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Formulation and Evaluation of Polyherbal
Formulations for Canker Sore (Mouth Ulcer)
A
Dissertation
Submitted for the Degree of
MASTER OF PHARMACY
IN
THE FACULTY OF PHARMACY
(PHARMACOGNOSY)
TO
GANPAT UNIVERSITY, KHERVA
2008- 2009
Research Guide: Submitted by:
Mrs. Nikunjana Patel Karuna J. Patel
M. Pharm. B. Pharm.
S. K. PATEL COLLEGE OF PHARMACEUTICAL EDUCATION AND RESEARCHGANPAT UNIVERSITY, KHERVA-382711.
DIST – MEHSANA (GUJARAT), INDIA.
CERTIFICATECERTIFICATEI hereby certify that Miss Karuna J. Patel has completed her thesis for Master of
Pharmacy on the topic “FORMULATION AND EVALUATION OF
POLYHERBAL FORMULATIONS FOR CANKER SORE (MOUTH
ULCER)”. I further certify that the work done by her is of her own and original and
tends to the general advancement of knowledge. The work was carried out under my
supervision and guidance at Department of Pharmacognosy, S. K. Patel College of
Pharmaceutical Education and Research, Ganpat Vidyanagar, during the
academic year 2008-2009. This work is up to my satisfaction.
Research Guide:
Mrs. Nikunjana PatelM. Pharm.Assistant Professor, Department of Pharmacognosy.
Forwarded Through:
Dr. N. J. Patel
M. Pharm., Ph.D.,Principal,
S.K.Patel College of Pharmaceutical Education and Research,Ganpat University, Kherva-382711.
Date:
Place: Ganpat University
Head of Department:
Dr. R. K. PatelM. Pharm., Ph.D.Head & Associate Professor,Department of Pharmacognosy.
DECLARATIONDECLARATION
I, Karuna J. Patel hereby declare that the topic entitled “FORMULATION
AND EVALUATION OF POLYHERBAL FORMULATIONS FOR CANKER
SORE (MOUTH ULCER)” which is submitted to the Ganpat University, Kherva, in
partial fulfilment for the award for Degree of Master of Pharmacy in
Pharmacognosy. The result of the work done by me in Department of Pharmacognosy
under the guidance of Mrs. Nikunjana Patel, Assistant Professor S. K. College of
Pharmaceutical Education and Research, Ganpat University, Kherva. I further declare
that the results of this work have not been previously submitted for any degree of
fellowship.
Place: Ganpat University. Karuna J. Patel
Date: - (B. Pharm.) Department of Pharmacognosy, S.K.P.C.P.E.R. Ganpat University
ACKNOWLEDGEMENTACKNOWLEDGEMENTIt is great pleasure for me to acknowledge all those who have contributed towards the
conception, origin and nurturing of this project. This work was carried out at the
Department of Pharmaceutics and Pharmaceutical Technology, S. K. Patel College
of Pharmaceutical Education and Research, Kherva during the years 2008-2009
which affiliated with Ganpat University, Kherva.
With a deep sense of gratitude and the respect, I thank my esteemed guide and
preceptor Mrs. Nikunjana Patel, for her inestimable guidance, valuable suggestions
and constant encouragement during the course of this study. It is with affection and
reverence that I acknowledge my indebtness to her and her outstanding dedication,
often far beyond the call of duty. Apart from guiding me, her unwiring moral support
and advice.
I take this opportunity to place on record my indebtness to Dr. Rakesh K.
Patel ,Head & Associate Professor ; I take this opportunity to say many thanks to Mr.
Hardik Patel. Mr. Kapil Khambholja, Lecturers, Department of Pharmacognosy for
their valuable suggestions, directions and selfless support throughout the
investigation.
I also expressed my profound gratitude to Dr. N. J. Patel, Principal & H.O.D,
Department of Pharmacology and toxicology, S. K. Patel College of Pharmaceutical
Education and Research, Kherva who have been a constant source of inspiration to
steer me forward through the two years of study and also for allowing me to use
facility during my project work..
I special mention thanks to Dr. S .S. Pancholi, H.O.D, Department of quality
assurance, and other faculty members of quality assurance, for their valuable
suggestions, directions and selfless support throughout the investigation.
I am also thankful to Mr. H. R Patel, Lecturer, Department of Pharmaceutics, for his
valuable help in my project work.
I sincerely thank to Mr. P. I. Patel, Librarian and assistant Mahadevbhai and
Mukeshbhai for providing me library facilities and their untiring and really
appreciable help in my literature survey.
I would like to express my thanks to non-teaching staff especially, Rakeshbhai (Lab.
Assistant), Jaimin bhai and Madhuben for their untiring help and support
throughout my M. Pharm. course.
I also thankful to Mr. Manishbhai Patel, Mrs. Kiranben Patel, (Lab. Assistant.
pharmaceutics department), Mr. Dineshbhai Patel (Store in charge), Mr. Sushilbhai
Patel (Lab. Assistant. quality assurance department), Mr. Nrupeshbhai Patel and
Mr. Rajubhai Raval (office staff) for their support throughout my M. Pharm. Course.
I sincerely acknowledge the help rendered to me by colleagues of M. pharm, Megha,
Sangita, Abhilasha, Monika, Pathik, Kaushik, Asif, Piyush and Umang for being co-
operative. Without their constant support and ideas this work would have been
difficult for me.
I equally thankful to my junior colleagues Tejas, Jigar, Nirav, Nevil, Rahul, Priya,
Kajal, Urvashi , Riddhi and Sejal for kind co-operation and timely help in the
fulfillment of my course work.
Memories play an important part in keeping special people close at heart. I can not
forget the sweat memories of the time that I have spent with my friends Manish,
Nisha, Anita and Ripal, who contributed in one or another way knowing and
unknowingly to reach out giving an encouragement and constant support when I am
need.
I hearty expressed my deep gratitude to my parents Mr. Jayantibhai Patel and Smt.
Sushilaben Patel, my brother Devendra, and other family members for their moral
support, constant encouragement and patience absolutely needed to complete my
entire study. They never opposed me and allowed me to what I want to do. They gave
me all facility and support in every situation of life. Taught me key of successes and
how to become a good person in life. Without their support I was not able to reach at
this level. It was the blessing of them that gave me courage to face the challenges and
made my path easier.
Thank you papa and mom.
Thank you very much to every body.
Karuna J. Patel
Dedicated
to
my beloved
family
Chapter.1Introduction
Chapter.2Review
ofLiterature
Chapter.3Aim and Plan of the
work
Chapter.4Evaluation of Raw
materials
Chapter.5Formulation and
Evaluation ofGel
Chapter.6Formulation and
Evaluation ofPatch
Chapter.7Antimicrobial activity
ofselected formulations
Chapter.8Summary
andConclusion
List ofTables and Figures
Index
List of Tables
List of Tables
Table
No.Title Page No.
1.1 Physical and Chemical Properties of Carbopol 28
1.2 Viscosity range of different Carbopol Polymers 28
4.1 Physicochemical parameters of G. glabra powder 70
4.2Observation table for calibration curve of std. 18-β- G.A.
(Using HPTLC method)72
4.3 Physicochemical parameters of A. catechu powder 74
4.4Observation table for calibration curve of std. catechin
(Using HPTLC method)76
4.5 Results of performed physical parameters of clove oil 77
4.6Observation table for calibration curve of std. eugenol
(Using HPTLC method)79
5.1 Preparation of gel formulations 83
5.2 Evaluation Parameters of Gels 87
5.3 In vitro Release profile of gel formulations 90
6.1 Preparation of patch formulations 94
6.2 Evaluation Parameters of Patches 98
6.3 In vitro Release profile of patch formulations 101
7.1 Zone of Inhibition (cm) of gel formulations 106
7.2 Zone of Inhibition (mm) of patch formulation 107
M. Pharm. (Pharmacognosy) I
List of Figures
List of Figures
Figure
No.Title
Page
No.1.1 Glycyrrhiza glabra Plant and Root 11.2 Acacia catechu Plant and Extract 61.3 Syzygium aromaticum plant , flower buds and oil 111.4 Types of Intraoral dosage forms 171.5 Variation in buccal patches 231.6 General Structure of Carbopol Polymers 261.7 Schematic drawing of a molecular segment of a cross-
linked polyacrylic acid polymer
27
1.8 Schematic representation of Hypromellose 341.9 Mouth Ulcer 404.1 Morphology of Glycyrrhiza glabra 684.2 Microscopic characters of G. glabra powder sample 684.3 TLC profile of Glycyrrhiza glabra powder 694.4 HPTLC profile of G. glabra powder 714.5 HPTLC chromatogram of standard 18-β-G.A. 714.6 HPTLC chromatogram of G. glabra extract 724.7 Calibration curve of 18-β- G.A. by HPTLC method 724.8 Morphology of A. catechu 734.9 TLC profile of A. catechu powder 744.10 HPTLC profile of A. catechu powder 754.11 HPTLC chromatogram of standard catechin 754.12 HPTLC chromatogram of standard A. catechu extract 764.13 Calibration curve of catechin by HPTLC method 764.14 TLC profile of clove oil 784.15 HPTLC profile of clove oil 784.16 HPTLC chromatogram of standard Eugenol 794.17 HPTLC chromatogram of clove oil 794.18 Calibration curve of Eugenol by HPTLC method 805.1 Prepared Gel Formulation 865.2 Calibration curve of 18-β-G.A. by HPTLC method 865.3 Calibration curve of Catechin by HPTLC method 875.4 HPTLC profile of Gel 885.5 HPTLC chromatogram of gel 885.6 HPTLC chromatogram of standard 18-β-G.A. 895.7 HPTLC chromatogram of standard catechin 895.8 HPTLC chromatogram of standard Eugenol 895.9 In vitro release profile of 18-β-G.A. in gel in Phosphate
buffer pH 6.8 at 254 nm
90
5.10 Calibration curve of Catechin by HPTLC method 916.1 Prepared Patch Formulation 976.2 Calibration curve of 18-β-G.A.by HPTLC method 976.3 Calibration curve of Catechin by HPTLC method 986.4 HPTLC profile of Patch 99
M. Pharm. (Pharmacognosy) II
List of Figures
6.5 HPTLC chromatogram of Patch 996.6 HPTLC chromatogram of standard 18-β-G.A. 1006.7 HPTLC chromatogram of standard Catechin 1006.8 HPTLC chromatogram of standard Eugenol 100
6.9In vitro release profile of 18-β-G.A. in patch in phosphate
buffer pH 6.8 at 254 nm
101
6.10In vitro release profile of catechin in patch in phosphate
buffer pH 6.8 at 254 nm
102
7.1 Antimicrobial Activity of gel formulation 1057.2 Antimicrobial Activity of patch formulation 106
M. Pharm. (Pharmacognosy) III
Index
Index
Chapter
NoTitle
Page
No.
List of Tales
List of Figures
I
II
1 Introduction 1-421.1
1.2
1.3
1.4
Introduction of Plants
Introduction of Buccoadhesive Drug Delivery
Introduction of Polymers
Introduction of Mouth Ulcer
References2 Review of Literature 43-58
2.1
2.2
2.3
2.4
2.5
Review Literature on Glycyrrhiza glabra and
Glycyrrhizin
Review Literature on Acaia Catechu and Catechin
Review Literature on Clove and Eugenol
Review Literature on Buccoadhesive Drug Delivery
Review Literature on Mouth Ulcer
References
3 Aim and Plan of the work 594 Evaluation of Raw materials 60-81
4.1
4.1.1
4.1.2
4.1.3
4.1.4
4.1.5
4.1.6
4.2
4.3
Experimental
Procurement of Raw Materials
Evaluation of G. glabra Whole Material and Powder
Evaluation of A. catechu Whole Material and Powder
Evaluation of Clove oil
Preparation of Extracts
Standardization of Extracts by HPTLC
Results and Discussion
Conclusion
References
5 Formulation and Evaluation of Gel82-92
M.Pharm. (Pharmacognosy)
Index
5.1
5.1.1
5.1.2
5.1.3
5.1.4
5.1.5
5.2
5.3
Experimental
Materials
Instruments
Preparation of Gel Formulations
Preparation of Standard Calibration Curves
Evaluation of Gel Formulations
Results and Discussion
Conclusion
References
6 Formulation and Evaluation of Patch93-103
5.1
5.1.1
5.1.2
5.1.3
5.1.4
5.1.5
5.2
5.3
Experimental
Materials
Instruments
Preparation of Patch Formulations
Preparation of Standard Calibration Curves
Evaluation of Patch formulations
Results and Discussion
Conclusion
References
7 Antimicrobial Activity of Selected Formulations 104-107
7.1
7.2
7.3
7.4
Introduction
Material and Method
Results and Discussion
Conclusion
References8 Summary and Conclusion 108
M.Pharm. (Pharmacognosy)
Chapter: 1.1 Introduction of Plants
A. Glycyrrhiza glabra
Source 1
Drug consists of the dried roots of Glycyrrhiza gtabra Linn. (syn. Liquiritae
officinalis Moench. ), Fam. Fabaceae. The plant is cultivated in Punjab and sub-
Himalayan tracts.
Figure: 1.1 Glycyrrhiza glabra plant and root
Classical Names 2
Yashtimadhu, Yashti. Yashtimadhuka, Madhuyashtika, Madhuka, Kleetaka,
Yashtyahva.
Vernacular Names 2
Eng.- Liquorice
Hindi- Mulhatii, Jethimadh, Jethimadhu, Mulelhi, Muleti, Mulathi
Beng.- Jashtimadhu. Jai-shbomodhu
Guj.- Jethimadha, Jethimard
Kan.- Yashti madhuka, Atimadhura, Jesthamadhu, Madhuka
Mal.-Yashtimadhukam. Atimadhuram, Irattimadhuram
Mar.- Jeshthamadh
Punj.- Mulelhi. Jethimadh
Tam.- Atimadhuram
Tel.- Yashtli-madhukam, Atimadhuramu
Arab.- Aslussus
Assam- Jesthimadhu, Yeshtmadhu
Burm.-Noe-khiyu, Noe-khiyu-asui
Kash.- Multhi
Urdu- Mulelhi, Asl-us-sus.
M. Pharm. (Pharmacognosy) 1
Chapter: 1.1 Introduction of Plants
Botanical Description 2
It is a hardy herb or undershrub attaining a height up to 2 m.; leaves multifoliolate,
imparipinnate; flowers in axillary spikes, papilionaceous, lavender to violet in colour;
pods compressed, containing reniform seeds.
Ayurvedic Properties 2
Rasa - Madhura
Guna - Guru, Snigdha
Veerya - Sheeta
Vipaka – Madhura
Doshaghnata - Vatapittashamaka
Rogaghnata - Vranashotha, Visha, Khalitya, Palitya, Shastrabhighataja vrana,
Vatavikara, Vatarakta, Amavata, Shiroroga. Vamana, Trishna,
Vibandha,Udarashoola, Amlapitta, Paittika apasmara. Hikka, Raktavikara,
Raktalpata, Raktapitta, Shwasa, Kasa, Swarabheda, Yakshma, Urogata vrana,
Urahkshata. Parshwashoola, Mootrakrichchhra, Pooyameha. Paittika
prameha,Shukrameha, Varnavikara, Kandu, Charma roga, Jeerna jwara,
Samanya daurbalya, Netra roga
Karma – Dahashamaka, Keshya, vedanasthapana, Shothahara, Nadibalya,
Medhya, Chhardinigrahuna, Trishnanigrahana, Vatanulomana, Mridurechana,
Shonitasthapana, Kaphanissaraka, Kanthya, Mootrala, Mootruvirajaneeya,
Shukravardhaka,Varnya.,Kandughna,Jwarashamaka,Jeevaneeya,Sandhaneea,
Rasayna. Balya, Chakshushya
Parts Used 3
Root
Chemical Constituents 1
Major: Triterpenoid saponin glycyrrhizin (2-9 %), a mixture of potassium and
calcium salts of glycyrrhizinic acid
Minor: Include other triterpenoid saponons viz.,glabranin A&B, Glycerrhetol,
glabrolide, iso- glabrolide;viz., formononetin, glabrone, neoliquiritin, hispaglabridin
A&B; coumarins viz., herniarin, umbelliferone; triterpene sterols viz., onocerin, β-
amyrin, stigmasterol
M. Pharm. (Pharmacognosy) 2
Chapter: 1.1 Introduction of Plants
Glycyrrhizin
Quantitative Standards 1
Ash values
Total ash - Not more than 10.0 %
Acid insoluble ash - Not more than 2.5 %
Extractive values
Alcohol soluble extractive -Not less than 10.0 %
Water soluble extractive - Not less than 20.0 %.
Adulratants / Substitues 4
The root of Abrus precatorious Linn. are known in the trade of Indian liquorice. Root
and Rhizome of Glycyrrhiza uralensis Fisch. ex DC. and some related plant species
are used as substitutes and adulterants of liquorice. G. uralensis contains only a small
percentage of sugar and gives a rather pungent extract.
Pharmacology
The drug possesses potent demulcent, expectorant, and anti-inflammatory properties
and these are attributed to the presence of glycyrrhizin. The latter is 50 times sweeter
than sucrose 5. Glycyrrhizin is also credited with antihepatotoxic 5,antiviral and
antibacterial 6 activities.
The drug is also use in the treatment of peptic ulcer and deglycyrrhizinated liquorice
while being substantially free from mineralocorticoid side effects of liquorice root is
clinically effective for gastric and deuodenal ulcers. It also indicates that in addition to
glycyrrhizinic acid, other unidentified constituents of the drug contribute to the
antiulcer activity 7, 8.
M. Pharm. (Pharmacognosy) 3
Chapter: 1.1 Introduction of Plants
Clinical Studies 2
During a controlled clinical trial conducted on 92 cases of post-operative traumatic
inflammation following tonsillectomy, 28 cases were given Yashtimadhu powder in a
dose of 3 gm t.d.s. In another series of 24 cases, oxyphenbutazone 2 tablets t.d.s. were
given. On sequential analysis, the anti-inflammatory response of Yashtimadhu was
found to be equivalent to that of oxyphenbutazone. It appeared to possess a more
potent anti-pyretic and anti-exudative activity as compared to oxyphenbutazone.
Contra Indication 10
The German Commission E lists cholestatic liver disorders, liver
cirrhosishypertension, hypokalaemia, severe kidney insufficiency and pregnancy as
contraindications. It is also Contraindicated in oedema and congestive heart failure.
Potassium loss due to other drugs, like thiazide diuretics, can be increased. Thus they
should not be taken together with glycyrrhizin.
Therapeutic Category 7
Anti-inflammatory, Antiulcer
Safety Profile 1
The drug when used within the recommended dosage the treatment period is devoid
of any adverse reactions. However, if taken in excessive amounts it can cause
matabolic disturbances known as pseudoaldosteronism ( due to the mineralocortecoid
effect of glycyrrhizin) leading to oedema, hypertension and weight gain.
Toxicology 2
LD50 of glycyrrhizin and glycyrrhizin- thiamine HC1 in rats is reported to be 1.94 and
0.764 g/kg s. c. respectively. Liquiritoside a root flavonoside is a low toxic substance.
Consumption of liquorice 10-45 g. / day is reported to cause raised blood pressure,
together with a block of aldosterone/ renin axis and electrocardiogram changes.
Dose 3, 9
Powdered drug: 2-4 gm
Liquorice compound powder: 4-8 ml
Liquorice liquid extract: 2-4 ml
M. Pharm. (Pharmacognosy) 4
Chapter: 1.1 Introduction of Plants
Formulations and Preparations 2
Yashtyadi churna, Yashtimadhvadya taila, Kalyanavaleha, Angamardaprashamana
kashaya, Brihat ashwagandha ghrita, Brihachchhagaladya ghrita, Shatavaryadi
ghritla, Nasika churna, Guduchyadi taila, Pippalyadi taila, Vyaghri taila,
Kubjaprasarani tlaila, Vridhihara lepa.
References:
1. Anonymous, “Indian Herbal Pharmacopoeia”, Indian Drug Manufacture’s
Association, Mumbai 2002, 89-97.
2. Sharma P.C., Yelne M.B, Dennis T.J., Assisted by Joshi Aruna, Prabhune Y.S.,
Khade Kundan, Rawat M.S., Database of medicinal plants used in Ayurveda,
vol-III , 561-566.
3. Anonymous, “The Ayurvedic Pharmacopoeia of India”; Ministry of health &
family welfare, Department of Health, Govt. of India, Part I,1st ed.,vol-1,
1989,127.
4. Anonymous, “The wealth of India (Raw material)”, CSIR, New Delhi, 1956,
vol-IV, 152.
5. Chandler R.E., Can. Pharm. J., 1985,118,420.
6. Kiso Y., Johkin M., Hikino H., Hattori M., Sakamoto T., and Namba T., Planta
med., 1984,50, 298.
7. Kassir Z.A., Irish Med J., 1985,78,153.
8. Longman M.J.S., Drugs , 1977,14,105.
9. Anonymous, “Pharmacopoeia of India”, Ministry of Health, Gov. of India, 2nd
Edi. 1966,406.
10. Blumenthal M (editor), 1998. Complete German Commission E monographs:
Therapeutic Guide to Herbal Medicines. American Botanical Council,
Integrative Medicine Communications, Boston, Massachusetts.
M. Pharm. (Pharmacognosy) 5
Chapter: 1.1 Introduction of Plants
B. Acacia catechu
Source 1
Drug consists of either dried bark or dried aqueous extract known as Black catechu or
cutch, prepared from the heartwood of Acacia catechu Willd. (Syn. A. catechuoides
Wall.) : Fam. Fabaceae. Sub. fam. - Mimosaceae. The plant is distributed throghout
india mostly in dry types of forests. Mainly eastern slopes of Wastern ghats,
Andhrapradesh, Bihar, Punjab and Himalaya up to an elevation of 1500 m.
Figure: 1.2 Acacia catechu plant and extract
Classical Names 2
Khadira, Balpatra, Bahushalya, Dantadhawana, Raktasara, Yadniya, Gayatri.
Vernacular Names 2
Eng. - Cutch tree
Hindi – Khair
Beng. – Khayar
Guj. - Kherio baval
Khair, Kathe,Kher
Kan.- Kalu, Kachu, Kaggali, Kanti, Kaggal.nara, Kachinamara, Koggigida
Mal. - Karingali, Khadiram
Mar.- Kaderi, Khair
Punj.-Khair
Tam.- Karunkali, Kadiram, Karngalli
Tel.- Podalimanu,kaviri, Kachu, Kadiramu, Sandra
Assam- Kharira, Khara, Khayar, Khoria
Kash.-Kath
Urdu - Chanbe kaath
M. Pharm. (Pharmacognosy) 6
Chapter: 1.1 Introduction of Plants
Botanical Description 2
A moderate sized, deciduous tree, up to 3 m high. Leaves pinnate, with a pair of
recurved prickles at the base of the rachis. Flowers pale yellow, in cylindrical spikes.
Pods glabrous, oblong.
Ayurvedic Properties 2
Rasa - Tikta, Kashaya
Guna - Loghu, Ruksha
Veerya - Sheeta
Vipaka - Katu
Prabhava - Kushthaghna
Doshaghnata - Kaphapittashamaka
Rogaghnata - Aruchi, Atisara, Kaphaja kasa, Prameha, Kushtha, Twakaroga,
Jeernajwara, Raktapitta, Krimi.
Karma-Ruchivardhaka, Stambhana, Shonitasthapana. Mutrasangrahana,
Kushlhaghna, Kandughna, Vrana ropaka.
Parts Used 2
Bark, heartwood, catechu (kattha)
Chemical Constituents 3
Major: Tannins-condensed tannins : catechutannic acid(25-30%), catechin (2-12%),l-
epicaetchin, catechu red, gum (20-30%)
Minor: flavanoids flavanols- quercitrin, quercitin
Catechin
M. Pharm. (Pharmacognosy) 7
Chapter: 1.1 Introduction of Plants
Quantitative standards 1
Heartwood
Foreign matter: Not more than 2 %,
Ash values:
Total ash - Not more than 2 %
Acid insoluble ash - Not more than 0.2 %
Extractive values
Alcohol soluble extractive -Not less than 1 %
Water soluble extractive - Not less than 3 %.
Catechu
Ash values:
Total ash- Not more than 8 %
Acid insoluble ash- Not more than 0.5 %
Extractive values
Alcohol insoluble residue- Not more than 30 %
Water insoluble residue-Net more than 25 %
Adulratants / Substitutes 4
Catechu i.e. heart wood extract can be adultrated with mineral matter viz. Clay or
starch, etc. This can be easily detected by the higher percentage of total and acid
insoluble ash. Similar extract is also obtained from leaves and young shoots of
Uncaria Gambier Rox, Family Rubiaceae. It is known as pale catechu, which has a
pale colour, and lighter in weight. The Gambier fluorescin test can be applied to
distinguish the two. Black catechu does not respond to this test.
Pharmacology
It is useful in Passive diarrhea either alone or in combination with cinnamon or
opium.
The concentrated aqueous extract, known as khayer gum or cutch is astringent 5,
cooling and digestive 6, beneficial in cough and diarrhoea6, applied externally to
ulcers6, boils and eruptions of the skin6, and is used extensively in Ayurvedic
formulations.
M. Pharm. (Pharmacognosy) 8
Chapter: 1.1 Introduction of Plants
The extract known as catechu or cutch is used medicinally as an astringent in fevers
and other maladies 6.
The bark, in combination with other drugs, is prescribed for snake bite 5.
Hypoglycemic activity of Acacia catechu (seeda) was observed when administered to
normal albino rats7.
A. catechu shows hypotensive action 8.
Agglutinins from seeds are administered to leukaemia patients 9.
A small piece of catechu with cinnamom and nutmeg is held in toothache, loss of
voice etc, also in cases of mercurial salivation, hoarseness, relaxed sore throat,
bleeding, ulcerations and sponginess of gums. It is used for bed sores 2.
Therapeutic category 11
Antiseptic, Astringent, Antidysentric
Safety profile 1
Usage of the drug in Indian system for centuries proves it’s safety. Both in acute and
sub- acute studies on the drug have found it safe. In higher doses, hemolytic action
may be observed.
Dose 10
Crude: 3-10 gm
Dried extract: 100-340 mg
Tincture (1:5): 2.5-5 ml
Formulations and Preparations2
Khadirashtaka churna, Kadhiradikvalha, Khadiradi vati, Arimedadi taila,
Khadirarishta
M. Pharm. (Pharmacognosy) 9
Chapter: 1.1 Introduction of Plants
References:
1. Anonymous, “Indian Herbal Pharmacopoeia”, Indian Drug Manufacture’s
Association, Mumbai 2002, Page no.1-11.
2. Sharma P.C., Yelne M.B, Dennis T.J., Assisted by Joshi Aruna, Prabhune Y.S.,
Khade Kundan, Rawat M.S., Database of medicinal plants used in Ayurveda, Vol-
I , 216-218.
3. Anonymous , “The wealth of India (Raw material)”, CSIR, New Delhi, 1985,vol-
I: A,25
4. Shah C.S and Quadry J.S., A Textbook of Pharmacognosy, B.S. Shah Prakashan,
Ahmedabad,7th edition,1990,154,157,158.
5. Anonymous, “British Pharmacopoeia”, Department of Health, British
Pharmacopoeia commission, London 1999.
6. Kirtikar K.R., Basu B.D., “Indian medicinal plants”.2nd edi., L. M. Basu,
Ahmadabad, Vol. II,926,927.
7. Singh K.N., Mittal R.K., and Barthwal K.C., et al.; Indian J Med Res, 1976,
64,754.
8. Sham K. N., Chiu K. W., Hypotensive Action Of Acacia Catechu, Planta Med,
1984, 50,177.
9. Agrawal S. and Agrawal S.S. et al; Indian J Med Res, 1990, 92: 38-42.
10. Chaudhari R.D., Herbal Drug Industry, Eastern publishers New Delhi, 1996,321.
11. Nadkarni K.M., Indian Materia Medica, Popular Prakashan, Bombay, Vol-I,
1976,11-13.
M. Pharm. (Pharmacognosy) 10
Chapter: 1.1 Introduction of Plants
C. Syzygium aromaticum
Source 1
Drug consists of flower buds of Syzygium aromaticum (Linn.) Merrill & Perry [Syn.
Eugenia aromatica Baill; Eugenia caryophyllata Thumb ; Caryophyllus aromaticus
Linn.] Family. Myrtaceae, A tree indigenous to the Molucca islands, grown in India
in certain parts of Tamilnadu (The Nilgiris and Kanniyakumari) and Kerala
(Kottarakara and Chengannur).
Figure: 1.3 Syzygium aromaticum plant , flower buds and Oil
Classical Names 2
Lavanga, Devakusuma, Shriprasuna, Chandanapushpaka, Varija.
Vernacular Names 2
Eng. - Clove tree. Cloves
Hindi- Lavanga, Laung, Lavamg, Laumg
Beng.-Lavang, Langa
Guj.- lavang, Laving
Kan.- lavanga
Mal.- Karampu, Karayampoovu, Grampu, Chanki, Karayampu,Krambu
Mar.- Lvang, Lawangta
Punj.- Laung, Long, Karanfal
Tam.- Kiramhu, Lavangam, Kiramber, Ilavangap-pu, Karuvat-pui crambu
Tel.- lavangalu, Lavangamu, devekusumamu
Assam- lavang, Lan. Long
Kash.- Rung, Raung
M. Pharm. (Pharmacognosy) 11
Chapter: 1.1 Introduction of Plants
Botanical Description 2
A pyramidal or conical evergreen tree up to 12 m high. Leaves simple, lanceolate, in
pairs, 7-13 cm in length and 2-4 cm in breadth, gland dotted, fragrant. Flower buds
greenish to pink, aromatic, clustered at the ends of branches. Fruits fleshy, dark pink
drupes. Seeds grooved on one side, oblong.
Ayurvedic Properties 2
Rasa- Katui, 'T'ikta
Guna –laghu, teekshna
Veerya - Sheeta
Vipaka - Katu
Doshagnata - Kaphapittanhamaka
Rogaghnata- Shirahshoola, Pratishyaya, Mukharoga, Charmaroga, Amavata,
Katishoola, gridharasi, Dantashoola, Dhwajabhanga, Aruchi, Agnimandya, Ajeerna,
Adhmana, udarashoola. Arnlapilta. Chhardi, Trishna. Grahani,
Visuchika,Yakridvikara, Hridduaurbalya, Raktavikara, Kasa, Shwasa, Hikka,
Kshaya,Mootraikrichchhra, Jwara, Samanya daurbalya
Karma - Raktotkleshaka, Uttejaka, Krimighna, Deepana, Pachana, Ruchya,
Mukhadurgandhanashan, Vaishadyakar, Vatanulomana, Shoolaprashamana,
Shleshmahara, Shleshmaputihara, Shwasahara, Vajikarana. Stanyajanana,
Stanyashodhana, Mootrajanana, Amapachana, Jwaraghna, katupaushtika,
Chakshushya, Vranaropana, Vranashodhana
Parts Used 3
Flower buds, oil
Chemical constituents 1
Major: volatile oil (15-20 %) containing chiefly a phenol eugenol (55-85%) and β-
caryophyllene (10-20%)
Minor: Eugenol acetate and derivative of β-caryophyllene, α-humulene and its
epoxide, acetophenone, benzyl salicylate, α-cardinol, γ-decalactone, fenchone,
hexanal, 2-hexanone, methyl palmitate, γ-muurolene, palustrol, propylbenzoate and
α-thujene.
M. Pharm. (Pharmacognosy) 12
Chapter: 1.1 Introduction of Plants
Eugenol
Quantitative standards 1
Foreign matter: Not more than 4.0%
Ash values
Total ash - Not more than 7.0 %
Acid insoluble ash - Not more than 0.5 %
Extractive values
Alcohol soluble extractive -Not less than 15.0 %
Water soluble extractive - Not less than 14.0 %
Volatile oil: Not less than 15.0%
Adulratants/substitues4
Cloves are sometimes adulterated with mother-of-cloves, clove stems, exhausted
cloves, withered cloves, clove dust containing broken stamens and flowers,
farinaceous products, cereal starches, ground fruit-stones and unripe fruits of
Cinnamomum verum J.S. Presl.
Pharmacology
Clove has a positive effect on the healing of stomach ulers5.
The sesquiterpenes of the drug are known to posses activity on inducing the
detoxifying enzyme glutathione S-transferase in mouse liver and small intestine and
the ability of natural anticarcinogens to induce such detoxifying enzymes correlates
well with their ability to inhibit chemical carcinogenesis4, 6.
The oil is antiseptic, local anaesthetic, rubefacient and useful in catarrh, cough,
bronchitis, flatulence, colic, skin diseases, dyspepsia, vomiting, odontalgia, dental
caries and cephalalgia2.
Clinical studies2
An Ayurvedic toothpaste prepared from Zingiher officinale, Terminalia chebulu.
Acacia catechu, Piper nigrum, Syzygium aromaticum, Cimiamomum zeylanicum and
M. Pharm. (Pharmacognosy) 13
Chapter: 1.1 Introduction of Plants
camphor etc. has been investigated in 50 patients suffering from dental diseases. The
paste was highly effective in controlling the diseases.
Therapeutic category1
Carminative, stomachic
Safety profile7
Clove buds are devoid of any major side effects/toxicity. However the clove oil
should be used with caution orally and should not be used on the skin.
Toxicology 2
Essential oil from flower buds showed typical toxicity to cowpea weevil. With 1%
dose, complete mortality was recorded within 24 hours. However some mortality was
reported in 4 days at 0.75% dose.
Dose 7
Clove: 120-300 mg
Clove oil: 0.05-0.2 ml
Formulations and Preparations 2
Dashanasanskwachurna, Lavangadi churna, Brihat chandrodayamakaradhwaja,
Ashwagandhadi churna, Kumariasava, Jeerakarishta Shrikhandasava,
Saubhagyashunthi, Lavangadi vati, Lavanga chatuhsama, lavangodaka, Avipattikara
churna, Devakusumadi rasa.
M. Pharm. (Pharmacognosy) 14
Chapter: 1.1 Introduction of Plants
References:
1. Anonymous , “Indian Herbal Pharmacopoeia”, Indian drug manufacture’s
Association, Mumbai 2002,Page no.89-97
2. Sharma P.C., Yelne M.B, Dennis T.J., Assisted by Joshi Aruna, Prabhune Y.S.,
Khade Kundan, Rawat M.S., Database of medicinal plants used in Ayurveda, Vol-
IV , 358-362.
3. Anonymous, “The Ayurvedic Pharmacopoeia of India”; Ministry of Health &
Family Welfare, Department of Health, Govt. of India, Part I, 1st ed., Part- I, vol.
I, 1989, 80-81
4. Bisset N.G. Herbal Drugs and Phytopharmaceuticals, Medpharm, Stuttgart,
1994,130
5. Zaidi S.H., Singh G.B., and Khanna N.M., Indian J Med Res, 1958, 46,732.
6. Zheng G.Q., Kenney p.m., AND Lam L.K.T., J. Nat.Prod. 1992,55,999.
7. Newall C.A., Anderson L.A., and Phillipson J.D., Herbal Medicines, The
Pharmaceutical Press, London, 1996, 79.
M. Pharm. (Pharmacognosy) 15
Chapter: 1.1 Introduction of Plants
M. Pharm. (Pharmacognosy) 16
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
1.2 Introduction of Buccoadhesive Drug Delivery
The oral route is the most preferred route of drug delivery as it is convenient,
inexpensive, and versatile. However, drug delivery by this route has certain
disadvantages such as first-pass metabolism by the liver and gastrointestinal
enzymatic degradation of the drug. Therefore, other transmucosal routes such as
nasal, rectal, vaginal, ocular, and oral mucosa are being considered as alternatives to
conventional oral dosage forms for drug delivery to avoid the above disadvantages
associated with conventional oral delivery (i.e., tablets, capsules, syrups, etc.). Of
these routes of delivery, the buccal oral mucosa has emerged as one of the target sites
for administration of drugs in a wide variety of dosage forms, particularly for those
drugs targeted for local delivery in the oral cavity and systemic absorption.
The buccal route of drug delivery provides the opportunity for drug absorption
through the buccal epithelial lining of the oral cavity (mucosa of the cheek) for it to
exhibit its action locally or systemically. The noninvasive nature of administration,
ease and convenience of administration, precise localization, and increased
permeability of the buccal mucosa compared to other transepithelial routes makes this
a promising route of delivery. Also, the rich supply of blood vessels and lymphatics in
the buccal mucosa results in rapid onset of drug action for those that have the
requisite physicochemical profile 1-3.
Drugs absorbed from the buccal mucosa may directly enter the systemic
circulation by way of the jugular vein, minimizing the first-pass liver metabolism and
gastric acid- or enzyme-mediated degradation (salivary fluid has lower enzymatic
activity than gut). The presence of food or variations in the gastric emptying rate has
little or no influence on drug delivery by the buccal route. The continuous exposure of
the oral mucosal tissues to a multitude of substances and its high cellular turnover rate
makes the buccal tissue robust and less prone to local toxicity or irritation from drugs,
their dosage forms, and formulation excipients. The absence of Langerhans cells in
the oral mucosal tissues imparts tolerance to potential allergens 4. Therefore,
prolonged administration of drugs (chronic use) to the oral cavity is less prone to
induction of local tissue sensitization and allergic reactions. When compared to other
mucosal delivery routes, buccal drug delivery offers a higher degree of control and
reproducibility; it also allows the removal of the systemic dosage form (i.e., chewing
gums, lozenges, patches, etc.) to terminate drug absorption if necessary.
M. Pharm. (Pharmacognosy) 16
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
Figure 1.4: Types of Intraoral dosage forms
1.2.1 Advantages of Oral Mucosal Drug Delivery 5, 6
The rationale and key advantages of drug delivery to the oral cavity for local and
Systemic absorption includes:
1. Alternative to injection
2. Improved patient compliance, “patient-friendly”
3. Convenience
4. Targeted delivery to treat local diseases of the oral cavity
5. Potential route for delivery of proteins/peptides/vaccines
6. Opportunity for product line extension of quick-dissolving dosage forms
7. Dosing or administration anywhere at anytime without water
8. Increase drug bioavailability in some cases
1.2.2 Disadvantages of Oral Mucosal Drug Delivery 5, 6, 7
There are, however, a number of technical challenges to overcome in order to
effectively deliver drugs to the oral cavity for local oral mucosal absorption and
systemic delivery. The absorption of drugs from the oral mucosal tissues has the
following disadvantages.
1. Drug transport is by passive diffusion, drug absorption is low, which results in a
low bioavailability.
M. Pharm. (Pharmacognosy) 17
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
2. Buccal mucosa, like the small intestine, offers a lipoidal barrier and this route is
usually practical for small lipophilic molecules.
3. Only a few hydrophilic drugs or compounds such as certain amino acids and
monosaccharides have been reported to be transported via a carrier-mediated process.
4. Taste masking may be necessary for drugs that are bitter or irritable.
5. The total area for drug absorption is small (100 to 170 cm2) when compared to the
total area of gastrointestinal absorption.
6. The dosage form must be kept in place for effective absorption because salivary
flow may wash away the dissolved drug and the dosage form may be swallowed prior
to drug dissolution.
1.2.3 Intraoral Drug Delivery
For transdermal drug delivery, the rate-limiting barrier for a compound to be
absorbed into the systemic circulation is the stratum corneum, which is the keratinized
layer of the skin. Because the lining mucosa of the oral cavity is not keratinized, is
thinner (100 to 500 μm), and more permeable than skin, the degree of resistance for a
drug to penetrate the lining mucosal membrane is much less than that of skin. Similar
to the epithelium of the GIT and the skin, the epithelial lining mucosa is composed of
a constantly renewing cell population, which is produced by cell division in the basal
region.
Intraoral drug delivery overcomes hepatic first-pass metabolism and promotes
rapid systemic delivery with improved bioavailability with selected drugs having the
required physiochemical and biopharmaceutical characteristics. The oral mucosa
provides accessibility to allow for the precise localization of the dosage form for
targeted drug delivery. It also provides the opportunity to directly modify tissue
permeability, inhibit protease activity, or decrease immunogenic responses to drugs.
Therefore, the oral mucosa has emerged as one of the target sites for administration of
drugs in a wide variety of dosage forms, particularly for those drugs targeted for local
delivery in the oral cavity and those that required systemic absorption as well.1-3
Due to the relatively short cellular turnover time of the buccal mucosa, a
properly designed buccal adhesive delivery system may remain in place, under the
best circumstances, for 1 to 24 hours.
Historically, lozenges and troches, which are sucked on by the patient, and
sublingual tablets were the traditional means for delivering drugs to the oral cavity.
M. Pharm. (Pharmacognosy) 18
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
More recently, quick-dissolve dosage forms have become more popular, and
bioadhesive tablets and gels have been used to provide a longer contact time with the
absorbing tissue and thus minimize drug loss through clearance by salivary outflow.
Drug delivery via the membranes of the oral cavity can be subdivided as follows:
A. Sublingual delivery, which is the administration of the drug via the sublingual
mucosa (the membrane of the ventral surface of the tongue and the floor of the
mouth) to the systemic circulation.
B. Buccal delivery, which is the administration of the drug via the buccal mucosa
(the lining of the cheek and area between the gums and upper and lower lips) to the
systemic circulation.
C. Periodontal, gingival, and odontal delivery, for the local treatment of conditions
of the oral cavity, principally aphthous ulcers, bacterial and fungal infections, and
periodontal disease. These oral mucosal sites differ greatly from one another in terms
of anatomy, permeability to an applied drug, and their ability to retain the delivery
system for the desired length of time.
1.2.7 Classification of Intraoral Dosage Forms
Dosage forms targeted for delivery to the intraoral cavity can be classified in terms of
their dissolution or disintegration kinetics as either quick-dissolving (QD), slow
dissolving (SD), or nondissolving (ND), which release the drug over a period of
seconds up to 1 minute, 1 to 10 minutes, and >10 minutes to hours, respectively.
These IODs may be intended to present the drug for local release in the mouth (i.e.,
drug dissolved in saliva) or to the GIT (i.e., drug released as microparticles but not
dissolved in saliva) for subsequent systemic absorption. The various IODs may
further be targeted for release and local topical absorption by the tissues in the oral
cavity for treatment of local diseases or for systemic absorption for treatment of
diseases remote from the site of application. The IODs may be further categorized as
noninvasive (i.e., QD formulations), semi-invasive (i.e., patches, periodontal
filaments, microparticles, needleless injectors), or invasive (i.e., injection via needles,
as used in dental anesthesia, or other drug delivery devices) depending upon their
interaction with mucosal tissues and the degree to which they physically breach the
membrane barrier to absorption.
M. Pharm. (Pharmacognosy) 19
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
A. Quick-Dissolving Delivery Systems
QD delivery systems undergo disintegration or dissolution in the saliva, generally
within a few seconds to a minute, releasing the drug and inactive ingredients into the
oral cavity the major fraction of the drug will eventually be swallowed with the saliva
and transported along the GIT where the drug is subsequently absorbed. The technical
advantages of these dosage forms include: ease of swallowing, administration without
water anywhere and anytime, quick onset of action with some drugs, supervised
administration, buccal or sublingual absorption, and local therapy of the oral mucosa.
Therapeutic benefits of the mouth-dissolving dosage forms for patients may include:
enhanced efficacy, improved convenience, and improved compliance. Pharmaceutical
companies may benefit from these dosage forms due to product differentiation, life-
cycle management, reduction of development costs, and outsourcing. The following
therapeutic categories have been reported to have market opportunities for QD
delivery systems: nonopioid analgesics, opioid analgesics, migraine headache, cough
and cold, allergy, gastrointestinal, cardiovascular, central nervous system, urology,
and other categories.
B. Slow-Dissolving Delivery Systems
SD intraoral delivery systems are generally dissolved in the oral cavity within 1 to 10
minutes and the following products are included in this category: chewable tablets,
sublingual tablets, “lollipops,” mucoadhesive tablets, and buccal tablets. Although
there are many commercial products on the markets in the first three categories, only
a few mucoadhesive or buccal tablets have recently been introduced (i.e., Striant ®).
However, there have been numerous reports of mucoadhesive tablets investigated in
the scientific and patent literature, and some products are in the R&D pipeline.
Numerous U.S. and foreign patents as well as patent applications have been published
on buccal drug delivery with the first appearing in the late 1960s involving various
mucoadhesive polymers. These polymers include but are not limited to polyacrylic
acid, hydroxypropylmethyl cellulose, polyvinylpyrrolidone, hydroxypropyl cellulose,
vinylpyrrolidone and its copolymers, polyethylene oxide, and sodium carboxymethyl
cellulose.
C. Nondissolving Delivery Systems
ND intraoral dosage forms do not dissolve entirely when placed in the mouth and can
provide for controlled drug delivery from 10 minutes to several hours, and up to a day
or longer under the best circumstances. Examples of ND intraoral delivery systems
M. Pharm. (Pharmacognosy) 20
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
include the following dosage forms: chewing gums, buccal and gingival patches,
periodontal fibers, and drug delivery devices. Medicated chewing gum technology
provides a new competitive advantage for product life-cycle management for mature
and novel pharmaceuticals. This delivery system technology adds a new benefit in
efficacy, convenience, and quality of life. Chewing gum formulations offer optimal
patient controlled dose titration, an appealing mouth feel and texture, controlled
release of active substances, and can be manufactured under cGMPs. The advantages
of drug delivery in a gum formulation include fast onset of action, doses in the mg
range, few side effects, drug release for up to 60 minutes, and reduced first-pass liver
metabolism compared to oral dosage forms. A medicated chewing gum delivery
system may be ideal for drugs having limited oral absorption in conventional tablets
(i.e., nicotine) and indications where a quick onset of action is desirable, such as
treatment of migraine, pain, allergy, angina, motion sickness, and smoking cessation
may benefit.
Technologies similar to transdermal patches have been developed and designed to
adhere to either the gingival or buccal mucosa. Mucoadhesive patches (Cydot ®) have
been developed and gingival local anesthetic patches have been commercialized
(Dentipatch ®) that can provide for drug delivery over a period of from 30 minutes to
24 hours depending upon the patch design and therapeutic indication. Other controlled
drug delivery systems have been developed for the treatment of periodontal disease in
the form of thin filaments and microparticles that are compacted into the periodontal
pocket to provide for prolonged delivery of antibiotics such as tetracycline (Actisite ®
for up to 10 days).
1.2.8 Oral Mucosal Drug Delivery Systems
Several bioadhesive dosage forms for oral mucosal drug delivery have been
developed and some are already commercially available. They are mostly in the form
of patches and tablets, and some are in the form of ointments, films, and powders. The
newest approach in this area has been the use of multifunctional polymers (e.g.,
polymers possessing both bioadhesive and phase change characteristics) and/or
polymers that are bioadhesive but also capable of being penetration enhancers, or
peptide stabilizers. The following section focuses on bioadhesive drug delivery
systems such as patches and tablets, as well as the potential application of phase
change polymers.
M. Pharm. (Pharmacognosy) 21
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
A. Bioadhesive Tablets
Bioadhesive tablets for oral mucosal drug delivery are usually based on an erodible
tablet 18. An example of this uses crosslinked hydroxypropyl cellulose (SynchronR) as
both a bioadhesive and a drug release regulator 19. The drug (nitroglycerin) is mixed
with the bioadhesive and then compressed into tablets. Due to adhesion of the
gradually eroding polymer to the buccal mucosa, this dosage form may remain in
place for three hours and the drug release profile showed zero-order release kinetics.
They claim that the patient may talk, drink, and even eat with the tablet in place.
A two-layered tablet with hydroxypropyl cellulose and carbopol 934 as the
bioadhesive layer and a lactose nonadhesive as the backing layer was designed20. The
role of the upper (lactose) layer is to prevent drug diffusing away from the absorption
site and to allow easy placement of the tablet in the mouth. The lower layer, which
constitutes an erodible bioadhesive, together with triamcinolone acetonide as an
active ingredient provides sustained release of the drug for the treatment of local
aphtous stomatitis.
A delivery system to deliver local anesthesia for toothache also proposed 21, 22. This
three-layer tablet consists of a core containing the drug and bioadhesives
(hydroxypropyl cellulose and Carbopol 934 as before).
B. Bioadhesive Patches
Although bioadhesive patches pose a relatively new technology to pharmacy,
they have developed very quickly with the recent development of bioadhesive
technology. Generally speaking, four different types of adhesive patches have been
studied for oral mucosal drug delivery, as shown in Figure 1.5.
In these cases, the adhesive polymer serves either as a drug carrier itself, or an
adhesive layer link between a drug-loaded layer and the mucosa, or a shield to cover a
drug-containing disc. The design of these patches provides either unidirectional or
bidirectional release of the drug. The size of such systems typically varies from 1 to
16 cm 2, depending on the specific purpose of the application. Usually, 1 to 3 cm2
patches are commonly used because of convenience and comfort. However, the
administration site is also a factor. Large-size patches can be administered at the
central position of the buccal mucosa, (i.e., center of the cheek), whereas the
sublingual and gingival sites require a rather small-sized patch.
M. Pharm. (Pharmacognosy) 22
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
Figure 1.5: Variation in buccal patches: (a) bidirectional release from adhesive
patch by dissolution or diffusion; (b) unidirectional release from patch
embedded in an adhesive shield: (c) bidirectional release from a laminated
patch; (d) unidirectional release from a laminated patch. M: mucosa; P: polymer
with peptide; D: drug depot; S: adhesive shield; A: adhesive layer; B:
impermeable backing layer 25
A variety of polymers can be used for oral mucosal patches. This includes
water-soluble and insoluble polymers of both ionic and nonionic types 18. With
soluble polymer systems, drug release is accompanied by dissolution of the polymer;
therefore the overall drug release rate and duration are determined by both polymer
dissolution and drug diffusion, whereas in a nonsoluble hydrogel system, drug release
follows fickian or nonfickian diffusion kinetics, depending on design. The duration of
mucosal adhesion of different bioadhesive patches varies from minutes to days
depending on the type of polymer used, its amount per patch, and additional factors
such as the drying technique used to prepare the patches. Adhesive patches consisting
of two ply laminates with an impermeable backing layer and a hydrocolloid polymer
layer containing the drug has evaluated 23. The polymers they investigated were
hydroxyethylcellulose, hydroxypropylcellulose, poly(vinylpyrrolidone) and
poly(vinylalcohol). Their in vivo results indicated the adhesion time for all the above
polymers on human buccal tissue is within the range of several minutes. A patch
consisting of a mucoadhesive basement membrane, a ratelimiting center membrane,
and an impermeable facing membrane has developed. The mucoadhesive
polycarbophil permits tight attachment to the buccal mucosal and allows the patch to
remain in place for approximately 17 hours in dogs and humans regardless of eating
and drinking. A bilayer mucoadhesive polymer system (Carbopol 934 and PVP). It
consists of a fast-release layer and a sustained-release layer to achieve sustained
release of a peptide drug has reported24. This system is claimed to adhere to
gingival/alveolar mucosa for over 24 hours.
M. Pharm. (Pharmacognosy) 23
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
References:
1. De Vries, M.E., Bodde, H.E., Verhoef, J.C., and Junginger, H.E. (1991).
Developments in buccal drug delivery. Crit. Rev. Ther. Drug Carrier Syst., 8(3),
271-303.
2. Shojaei, A. (1998). Buccal mucosa as a route for systemic drug delivery: A
review. J.Pharm. Pharmaceut. Sci., 1, 15-30.
3. Squier, C.A. (1991). The permeability of oral mucosa. Crit. Rev. Oral Biol. Med.,
2(1),13-32.
4. Bodde, H.E., de Vries, M.E., and Junginger, E.H. (1990). Mucoadhesive polymers
for the buccal delivery of peptides, structure-adhesiveness relationships. J.
Control. Release, 13, 225-231.
5. Washington, N., Washington, C., and Wilson, C. (2001a). Physiological
Pharmaceutics-Barriers to Drug Absorption (2d ed.), New York: Taylor and
Francis.
6. Washington, N., Washington, C., and Wilson, G.C. (2001b). Drug delivery to the
oral cavity or mouth. Biological Pharmaceutics: Barriers to Drug Absorption (pp.
37-58). New York: Taylor and Francis.
7. Shojaei, A.H., Berner, B., and Li, X. (1998). Transbuccal delivery of acyclovir (I):
In vitro determination of routes of buccal transport. Pharm. Res., 15(8), 1182-
1188.
8. Zhou, X.H. Overcoming enzymatic and absorption barriers to non-parenterally
administered protein and peptide drug, J. Control Rel. 29:239-252, 1994.
9. Gutniak, M.K., Larsson, H., Sanders, S.W., Juneskans, O., Holst, J.J., and Ahren,
B. GLP-1 Tablet in type 2 diabetes in fasting and post-prandial conditions,
Diabetes Care, 20:1874-1879, 1997.
10. Merkle HP, Anders R, and Wermerskirchen A. Mucoadhesive buccal patches for
peptide delivery. In: Lenaerts V, Gurny R, eds. Bioadhesive Drug Delivery
Systems. Boca Raton, FL: CRC, 1990:105-136.
11. Schor JM, Davis SS, Nigalaye A, and Bolton S. Susadrin transmucosal tablets.
Drug Dev Ind Pharm 1983; 9:1359-1377.
12. Nagai T, Machida Y, Suzuki Y, and Ikura H. Method and preparation for
administration to the mucosa of the oral or nasal cavity. U.S. Patent 4226848,
1980.
M. Pharm. (Pharmacognosy) 24
Chapter: 1.2 Introduction of Buccoadhesive Drug Delivery System
13. Nagai T and Machida Y. Mucoadhesive dosage forms. Pharm Int 1985; 6:196-
200.
14. Ishida M, Nambu N, and Nagai T. Mucosal dosage form of lidocaine for
toothache using hydroxypropyl cellulose and carbopol. Chem Pharm Bull 1982;
30:980-984.
15. Anders R and Merkle HP. Evaluation of laminated muco-adhesive patches for
buccal drug delivery. Int J Pharm 1989; 49:231−240.
16. Lee Y and Chien YW. Oral mucosa controlled delivery of LHRH by bilayer
mucoadhesive polymer systems. J Control Rel 1995; 37:251-261.
17. Merkle HP, Anders R, Sandow J, and Schurr W. Drug delivery of peptides: the
buccal route. In: Davis SS, Illum L, Tomlinson E, eds. Delivery Systems for
Peptide Drugs. New York: Plenum, 1986:159-176.
M. Pharm. (Pharmacognosy) 25
Chapter: 1.3 Introduction of Polymers
1.3.1 Carbopol
Carbopol polymers are polymers of acrylic acid cross-linked with polyalkenyl ethers
or divinyl glycol. They are produced from primary polymer particles of about 0.2 to
6.0 micron average diameter. The flocculated agglomerates cannot be broken into the
ultimate particles when produced. Each particle can be viewed as a network structure
of polymer chains interconnected via cross-linking 1.
Carbomers were first prepared and patented in 1957 2. Since then, a number of
extended release tablet formulations, which involve carbomer matrices, have been
patented3.
Carbomers readily absorb water, get hydrated and swell. In addition to its hydrophilic
nature, its cross-linked structure and its essentially insolubility in water makes
Carbopol a potential candidate for use in controlled release drug delivery system 4, 5.
1. Nonproprietary Names
BP: Carbomers
PhEur: Carbomera
USPNF: Carbomer
2. Description
Carbopol polymers are offered as fluffy, white, dry powders (100% effective).
The carboxyl groups provided by the acrylic acid backbone of the polymer are
responsible for many of the product benefits. Carbopol polymers have an average
equivalent weight of 76 per carboxyl group 6. The general structure
can be illustrated with fig. No.1.6.
Figure 1.6: General Structure of Carbopol Polymers
M Pharm. (Pharmacognosy) 26
Chapter: 1.3 Introduction of Polymers
Figure 1.7: Schematic drawing of a molecular segment of a cross-
linked polyacrylic acid polymer
Carbopol polymers are manufactured by cross-linking process. Depending upon the
degree of cross-linking and manufacturing conditions, various grades of Carbopol are
available. Each grade is having its significance for its usefulness in pharmaceutical
dosage forms7.
Carbopol 934 P is cross-linked with allyl sucrose and is polymerized in solvent
benzene. Carbopol 71G, 971 P, 974 P are cross-linked with allyl penta erythritol and
polymerized in ethyl acetate. Polycarbophil is cross-linked polymer in divinyl glycol
and polymerized in solvent benzene. All the polymers fabricated in ethyl acetate are
neutralized by 1-3% potassium hydroxide. Though Carbopol 971 P and Carbopol 974
P are manufactured by same process under similar conditions, the difference in them
is that Carbopol 971 P has slightly lower level of cross-linking agent than Carbopol
974 P. Carbopol 71 G is the granular form Carbopol grade8.
3. Physical Properties 9
The three dimensional nature of these polymers confers some unique characteristics,
such as biological inertness, not found in similar linear polymers. The Carbopol resins
are hydrophilic substances that are not soluble in water. Rather, these polymers swell
when dispersed in water forming a colloidal, mucilage-like dispersion.
Carbopol polymers are bearing very good water sorption property. They swell in
water up to 1000 times their original volume and 10 times their original diameter to
form a gel when exposed to a pH environment above 4.0 to 6.0. Because the pKa of
M Pharm. (Pharmacognosy) 27
Chapter: 1.3 Introduction of Polymers
these polymers is 6.0 to 0.5, the carboxylate moiety on the polymer backbone ionizes,
resulting in repulsion between the native charges, which adds to the swelling of the
polymer. The glass transition temperature of Carbopol polymers is 105°C (221°F) in
powder form.
Table 1.1: Physical and Chemical Properties of Carbopol 10
Appearance Fluffy, white, mildly acidic polymerBulk Density Approximately 208 kg/m3 (13 lbs.ft3) *Specific gravity 1.41Moisture content 2.0% maximumEquilibrium moisture
content
8-10% (at 50% relative humidity)
PKa 6.0 ± 0.5pH of 1.0% water
dispersion
2.5 - 3.0
pH of 0.5% water
dispersion
2.7 - 3.5
Equivalent weight 76 ± 4Ash content 0.009 ppm (average) **Glass transition temperature 100-105C (212-221F)
4. Rheological properties
While the relationships between structure and properties have been of interest both
academically and in industry. Different grades of Carbopol polymers exhibit different
rheological properties, a reflection of the particle size, molecular weight between
crosslinks (Mc), distributions of the Mc, and the fraction of the total units, which
occur as terminal, i.e. free chain ends 11-18.
Table 1.2: Viscosity range of different Carbopol Polymers 21, 22
Polymer Viscosity*Carbopol 934 NF 30500 – 39400
Carbopol 934 P NF 29400 – 39400Carbopol 71 G NF 4000 – 11000
5. Applications of Carbopol polymers
The readily water-swellable Carbopol polymers are used in a diverse range of
pharmaceutical applications to provide:
Controlled release in tablets 1, 23-25.
Bioadhesion in buccal 26, ophthalmic 27, 28, intestinal 29, nasal 30, vaginal 31 and
rectal 32 applications.
M Pharm. (Pharmacognosy) 28
Chapter: 1.3 Introduction of Polymers
Thickening at very low concentrations to produce a wide range of viscosities
and flow properties in topical, lotions, creams and gels, oral suspensions and
transdermal gel reservoirs 33.
Permanent suspensions of insoluble ingredients in oral suspensions and
topicals 34.
Emulsifying topical oil-in-water systems permanently, even at elevated
temperatures, with essentially no need for irritating surfactants.
6. Bioadhesive Applications 35
Bioadhesion is a surface phenomena in which a material may be of natural or
synthetic origin, adheres or stick to biological surface, usually mucus membrane. The
concept of bioadhesion is emerging as a potential application in drug delivery due to
its applicability for bioavailability enhancement, prolongation of residence time for
drug in GIT and better contact between drug and absorbing surface.
Many hydrophilic polymers adhere to mucosal surfaces as they attract water from the
mucus gel layer adherent to the epithelial surface. This is the simplest mechanism of
adhesion and has been defined as “adhesion by hydration” Various kinds of adhesive
force, e.g. hydrogen bonding between the adherent polymer and the substrate, i.e.
mucus, are involved in mucoadhesion at the molecular level. Carbopol polymers have
been demonstrated to create a tenacious bond with the mucus membrane resulting in
strong bioadhesion.
Many commercial oral and topical products available today and under investigation
have been formulated with Carbopol polymers, as they provide numerous benefits in
bioadhesive formulations.
Benefits in Bioadhesive Applications
Improve bioavailability of certain drugs.
Enhance patient compliance (fewer doses are needed per day)
Lower concentrations of the active ingredients can be used.
Provide excellent adhesion forces.
7. Oral Care Applications 36,37
Carbopol polymers impart several desirable characteristics to toothpaste formulations
like Viscosity, Yield Value, Low thixotropy and Clarity.
Imparting viscosity at very low concentrations to thicken a system is a primary
function of the polymers. Suspending abrasives and solid actives is accomplished
through the build of yield value at low polymer concentrations. The combination of
M Pharm. (Pharmacognosy) 29
Chapter: 1.3 Introduction of Polymers
Carbopol polymers’ ability to build yield value with low thixotropy provides for a
clean, non-stringing ribbon of toothpaste. From aesthetic and practical perspectives
this means that Carbopol toothpaste formulations are pumpable, leave minimal solids
residue on the tube rim, stand up well on the brush, and can be used in clear
formulations.
Benefits in Oral care Applications:
Efficient co-binders at low usage levels.
Suspending agents for non-soluble actives or excipients.
Thicken peroxide gel systems while maintaining product stability.
Compatible with commonly used formulation ingredients.
References:
1. At Florence, Pu, Jani: “Novel Oral-Drug Formulations-Their Potential in
Modulating Adverse-Effects” Drug Saf., 1994., 410(3), 233-266.
2. H P Brown, US Patent No. 2798053 (1957)
3. B F Goodrich Bulletin: “Sustained Release Patents using Carbopol Resin” ,B F
Goodrich Company, Cleveeland, OH, 1987.
M Pharm. (Pharmacognosy) 30
Chapter: 1.3 Introduction of Polymers
4. Carnali J O, Naser M S: “The Use of Dilute Solution Viscosity to Characterize
the Network Properties of Carbopo® Microgels,” Colloid& Polymer Science,
1992,270(2),183-193.
5. Garcia-Gonzalez N, Kellaway, I W, Blanco, Fuente H, Anguiano, Igea S,
Delgado, Charro B,Otero, Espinar F J, Mendez J: “Influence of
Glycerol Concentration and Carbopol Molecular Weight on Swelling and Drug
Release Characteristics of Metoclopramide Hydrogels” Int. J.Pharm.,1994,
104,107-113.
6. B F Goodrich Company Technical Literature: Carbopol Resin Handbook, 1991
7. Alexander P, Organic Rheological Additives, Mfg. Chem., 1986,57(10),81-84.
8. Noveon Company Technical Literature, 2005.
9. Martindale – The Complete Drug Reference, 33rd Edition 2002 (1499).
10. Handbook of Pharmaceutical Excipients, Washington DC, American
Pharmaceutical Association / Pharmaceutical Society of Great Britain, 1986,41-
42.
11. Lochhead R Y, Davidson J A, Thomas G M: “Polymers in Aqueous Media”,
1989,13-147.
12. Meyer R J, Cohen L, “The Rheology of Natural and Synthetic Hydrophilic
Polymer Solutions as Related to Suspending Ability,” J. Soc. Cosmetic Chemists,
presented November 1958, New York City, 1958.
13. Flory, Paul J., Principles of Polymer Chemistry, Cornell University Press.
14. Reloar L R G: “The Physics of Rubber Elasticity, Oxford University Press”, New
York, 1958.
15. Advances in Polymer Science, Vol. 109 & 110, Responsive Gels: I & II,
Springer-Verlag.
16. Peppas N A: “Characterization of Cross-linked Structure of Hydrogels,”
Hydrogels Med.Pharm. , 1986,1, 27-56.
17. Brannon-Peppas L, “Preparation and Characterization of Cross-linked Hydrophilic
Networks,” Sud. Polym. Sci. , 1990, 8, 45-66.
18. Hooper H H et al: “Swelling Equilibrium for Positively Ionized Poly(acrylamide)
Hydrogels,” Macromolecules, 23(4) , 1990, 1096-1104.
19. Taylor, Bagley: “Tailoring Closely Packed Gel-Particles Systems for Use as
Thickening Agents,” J. Appl. Polym. Sci. , 1977, 21, 113-122.
M Pharm. (Pharmacognosy) 31
Chapter: 1.3 Introduction of Polymers
20. Taylor, Bagley: “Rheology of Dispersions of Swollen Gel Particles,”J. Polym.
Sci.: Polymer Physics Edition, 1975, 13, 1133-1144.
21. Nae H N, Reichert W W: “Rheological Properties of Lightly Cross-linked
Carboxy Copolymers in Aqueous Solutions,” Rheologica Acta,31, 1992 ,(4),
351-360.
22. Ishikawa S et al: “Evaluation of the Rheological Properties of Various Kinds of
Carboxyvinyl polymer Gels,” Chem. Pharm. Bull, 1988, 36(6),2118-2127.
23. Choulis N H, Papadopoulos H, Choulis M: “Long Acting Methadone,”
Pharmazie, 1976, 31, H 7.
24. Durrani, Manzer J, Whitaker, Roy, Benner, Samuel C: “A Comparative Study of
Controlled Release Agents- I: Effects of Compression Force and Polymer
Concentration,” Amer. Assoc Pharm. Sci., (Nov.), 1992.
25. Perez-Marcos B, Gutierrez C, Gomez-Amoza J L, Martinez-Pacheco R, Souto C,
Concheiro A: “Usefulness of Certain Varieties of Carbomer in the Formulation of
Hydrophilic Furosemide Matrices,” Int. J. Pharm. , 1991, 67(2), 113-121.
26. Guo J H: “Investigating the Bioadhesive Properties of Polymer Patches for Buccal
Drug Delivery (Carbopol934P),” J. Control. Release. ,1994, 28(1), 272-273.
27. Davies N M, Farr S J, Hadgraft J, Kellaway I W: “Evaluation of Mucoadhesive
in Polymers Ocular Drug Delivery-II. Polymer-Coated Vesicles ” Pharm. Res. ,
1992,9(9), 1137-44.
28. Akiyama Y, Nagahara N, Hirai S, Toguchi H: “ In Vitro and In Vivo Evaluation
of Mucoadhesive Microspheres Prepared for the Gastrointestinal Tract Using
Polyglycerol Esters of Fatty Acids and a Poly(Acrylic Acid) Derivative,”
Pharm. Res. , 1995,12(3), 397-405.
29. ElHady S S A, Mortada N D, Awad G A S, Zaki NM, Taha RA, “Development of
insitu gelling and mucoadhesive Mebeverine Hydrochloride solution for rectal
administration” Saudi Pharm. J. , 2003,11(4), 59-169.
30. Al-Khamis K I, Davis S S, Hadcraft, J: “Microviscosity and Drug Release from
Topical Gel Formulations,” Parma. Res. , 1986, 3(4), 214-217.
31. Briede R H: “ Application of Carbomer water Gel 1%”, Pharm. Week,1983,
118(9), 170-174.
32. Berney B M, Deasy P B, “ Evaluation of Carbopol 934P as a suspending agent
for Sulfademidine suspensions” Int. J. Pharm. , 1979,3(2-3), 73-80.
M Pharm. (Pharmacognosy) 32
Chapter: 1.3 Introduction of Polymers
33. Anlar S, Capan Y, Hincal A., A: “Physico-Chemical and Bioadhesive Properties
of Polyacrylic Acid Polymers,” Pharmazie, 1993, 48(4), 285-287.
34. Chang H S, Park H, Kelly P, Robinson J R: “ Bioadhesive Polymers as Platforms
for Oral Controlled Release Drug Delivery-II: Synthesis and Evaluation of Some
Swelling, Water- insoluble Bioadhesive Polymers,” J. Pharmacol. Sci. 1985,
74, 229.
35. Smart J D: “Some Formulation Factors Influencing the Rate of Drug Release from
Bioadhesive Matrices,” Drug Devel. Ind. Pharm. , 1992, 18(2), 223-232.
36. Lehr C M, Bouwstra J A, Tukker J J, Verhoef A C, de Boer A G, Junginger, H E,
Breimer D D: “Oral Bioadhesive Drug Delivery Systems - Effects on G.I. Transit
and Peptide Absorption,” Pharm. Res., 7(9), Sep 1990 (Suppl.) PDD 7226, 1990.
37. Longer M A, Chang H S, Robinson J R: “Bioadhesive Polymers as Platforms for
Oral Controlled Drug Delivery- III: Oral Delivery of Chlorothiazide Using a
Bioadhesive Polymer,” J. Pharm. Sci. , 1985, 74(4), 406-411.
1.3.2 Hypromellose
1. Nonproprietary Names
BP: Hypromellose
JP: Hydroxypropylmethylcellulose
PhEur: Hypromellosum
USP: Hypromellose
M Pharm. (Pharmacognosy) 33
Chapter: 1.3 Introduction of Polymers
2. Synonyms
Benecel MHPC; E464; hydroxypropyl methylcellulose; HPMC; Methocel;
methylcellulose propylene glycol ether; methyl hydroxypropylcellulose; Metolose;
Tylopur.
3. Chemical Name and CAS Registry Number
Cellulose hydroxypropyl methyl ether [9004-65-3]
4. Empirical Formula and Molecular Weight
The PhEur 2005 describes hypromellose as a partly O-methylated and O-(2-
hydroxypropylated) cellulose. It is available in several grades that vary in viscosity
and extent of substitution. Grades may be distinguished by appending a number
indicative of the apparent viscosity, in mPa s, of a 2% w/w aqueous solution at 20°C.
Hypromellose defined in the USP 28 specifies the substitution type by appending a
four-digit number to the nonproprietary name: e.g., hypromellose 1828. The first two
digits refer to the approximate percentage content of the methoxy group (OCH3). The
second two digits refer to the approximate percentage content of the hydroxypropoxy
group (OCH2CH(OH)CH3), calculated on a dried basis. Molecular weight is
approximately 10 000–1 500 000.
5. Structural Formula
where R is H, CH3, or CH3CH(OH)CH2
Figure 1.8: Schematic representation of Hypromellose
6. Functional Category
Coating agent; film-former; rate-controlling polymer for sustained release; stabilizing
agent; suspending agent; tablet binder; viscosity-increasing agent.
7. Applications in Pharmaceutical Formulation or Technology
M Pharm. (Pharmacognosy) 34
Chapter: 1.3 Introduction of Polymers
Hypromellose is widely used in oral, ophthalmic and topical pharmaceutical
formulations.
In oral products, hypromellose is primarily used as a tablet binder, 1 in film-coating, 2–7
and as a matrix for use in extended-release tablet formulations 8–12. Concentrations
between 2% and 5% w/w may be used as a binder in either wet- or dry-granulation
processes. High viscosity grades may be used to retard the release of drugs from a
matrix at levels of 10–80% w/w in tablets and capsules.
Depending upon the viscosity grade, concentrations of 2–20% w/w are used for film-
forming
solutions to film-coat tablets. Lower-viscosity grades are used in aqueous film-coating
solutions, while higher-viscosity grades are used with organic solvents. Examples of
filmcoating materials that are commercially available include AnyCoat C, Spectracel,
and Pharmacoat.
Hypromellose is also used as a suspending and thickening agent in topical
formulations. Compared with methylcellulose, hypromellose produces aqueous
solutions of greater clarity, with fewer undispersed fibers present, and is therefore
preferred in formulations for ophthalmic use. Hypromellose at concentrations between
0.45–1.0% w/w may be added as a thickening agent to vehicles for eye drops and
artificial tear solutions.
Hypromellose is also used as an emulsifier, suspending agent, and stabilizing agent in
topical gels and ointments. As a protective colloid, it can prevent droplets and
particles from coalescing or agglomerating, thus inhibiting the formation of
sediments.
In addition, hypromellose is used in the manufacture of capsules, as an adhesive in
plastic bandages, and as a wetting agent for hard contact lenses. It is also widely used
in cosmetics and food products.
8. Description
Hypromellose is an odorless and tasteless, white or creamy-white fibrous or granular
powder.
10. Typical Properties
M Pharm. (Pharmacognosy) 35
Chapter: 1.3 Introduction of Polymers
Acidity/alkalinity: pH = 5.5–8.0 for a 1% w/w aqueous solution.
Ash: 1.5–3.0%, depending upon the grade and viscosity.
Auto ignition temperature: 360°C
Density (bulk): 0.341 g/cm3
Density (tapped): 0.557 g/cm3
Density (true): 1.326 g/cm3
Melting point: browns at 190–200°C; chars at 225–230°C. Glass transition
temperature is 170–180°C.
Moisture content: Hypromellose absorbs moisture from the atmosphere; the amount
of water absorbed depends upon the initial moisture content and the temperature and
relative humidity of the surrounding air.
Solubility: Soluble in cold water, forming a viscous colloidal solution; practically
insoluble in chloroform, ethanol (95%), and ether, but soluble in mixtures of ethanol
and dichloromethane, mixtures of methanol and dichloromethane, and mixtures of
water and alcohol. Certain grades of hypromellose are soluble in aqueous acetone
solutions, mixtures of dichloromethane and propan-2-ol, and other organic solvents.
Specific gravity: 1.26
Viscosity (dynamic): A wide range of viscosity types are commercially available.
Aqueous solutions are most commonly prepared, although hypromellose may also be
dissolved in aqueous alcohols such as ethanol and propan-2-ol provided the alcohol
content is less than 50% w/w. Dichloromethane and ethanol mixtures may also be
used to prepare viscous hypromellose solutions. Solutions prepared using organic
solvents tend to be more viscous; increasing concentration also produces more
viscous solutions.
To prepare an aqueous solution, it is recommended that hypromellose is dispersed and
thoroughly hydrated in about 20–30% of the required amount of water. The water
should be vigorously stirred and heated to 80–90°C, then the remaining hypromellose
should be added. Sufficient cold water should then be added to produce the required
volume.
When a water-miscible organic solvent such as ethanol (95%), glycol, or mixtures of
ethanol and dichloromethane are used, the hypromellose should first be dispersed into
the organic solvent, at a ratio of 5–8 parts of solvent to 1 part of hypromellose. Cold
water is then added
to produce the required volume.
M Pharm. (Pharmacognosy) 36
Chapter: 1.3 Introduction of Polymers
11. Stability and Storage Conditions
Hypromellose powder is a stable material, although it is hygroscopic after drying.
Solutions are stable at pH 3–11. Increasing temperature reduces the viscosity of
solutions. Hypromellose undergoes a reversible sol–gel transformation upon heating
and cooling, respectively. The gel point is 50–90°C, depending upon the grade and
concentration of material.
Aqueous solutions are comparatively enzyme-resistant, providing good viscosity
stability during long-term storage 13. The aqueous solutions are liable to microbial
spoilage and should be preserved with an antimicrobial preservative.
Hypromellose powder should be stored in a well-closed container, in a cool, dry
place.
12. Incompatibilities
Hypromellose is incompatible with some oxidizing agents. Since it is nonionic,
hypromellose will not complex with metallic salts or ionic organics to form insoluble
precipitates.
13. Method of Manufacture
A purified form of cellulose, obtained from cotton linters or wood pulp, is reacted
with sodium hydroxide solution to produce a swollen alkali cellulose that is
chemically more reactive than untreated cellulose. The alkali cellulose is then treated
with chloromethane and propylene oxide to produce methyl hydroxypropyl ethers of
cellulose. The fibrous reaction product is then purified and ground to a fine, uniform
powder or granules.
14. Safety
Hypromellose is widely used as an excipient in oral and topical pharmaceutical
formulations. It is also used extensively in cosmetics and food products.
Hypromellose is generally regarded as a nontoxic and nonirritant material, although
excessive oral consumption may have a laxative effect 14. The WHO has not specified
an acceptable daily intake for hypromellose since the levels consumed were not
considered to represent a hazard to health 15-16.
M Pharm. (Pharmacognosy) 37
Chapter: 1.3 Introduction of Polymers
15. Handling Precautions
Observe normal precautions appropriate to the circumstances and quantity of material
handled. Hypromellose dust may be irritant to the eyes and eye protection is
recommended. Excessive dust generation should be avoided to minimize the risks of
explosion. Hypromellose is combustible.
References:
1. Chowhan ZT. Role of binders in moisture-induced hardness increase in compressed
tablets and its effect on in vitro disintegration and dissolution. J Pharm Sci 1980;
69: 1–4.
2. Rowe RC. The adhesion of film coatings to tablet surfaces – the effect of some
direct compression excipients and lubricants. J Pharm Pharmacol 1977; 29: 723–
726.
3. Rowe RC. The molecular weight and molecular weight distribution of
hydroxypropyl methylcellulose used in the film coating of tablets. J Pharm
Pharmacol 1980; 32: 116–119.
4. Banker G, Peck G, Jan S, Pirakitikulr P. Evaluation of hydroxypropyl cellulose and
hydroxypropyl methyl cellulose as aqueous based film coatings. Drug Dev Ind
Pharm 1981; 7: 693–716.
5. Okhamafe AO, York P. Moisture permeation mechanism of some aqueous-based
film coats. J Pharm Pharmacol 1982; 34 (Suppl.): 53P.
6. Alderman DA, Schulz GJ. Method of making a granular, cold water dispersible
coating composition for tablets. United States Patent No. 4,816,298; 1989.
7. Patell MK. Taste masking pharmaceutical agents. United States Patent No.
4,916,161; 1990.
8. Hardy JG, Kennerley JW, Taylor MJ, et al. Release rates from sustained release
buccal tablets in man. J Pharm Pharmacol 1982; 34 (Suppl.): 91P.
9. Hogan JE. Hydroxypropylmethylcellulose sustained release technology. Drug Dev
Ind Pharm 1989; 15: 975–999.
10. Shah AC, Britten NJ, Olanoff LS, Badalamenti JN. Gel-matrix systems exhibiting
bimodal controlled release for oral delivery. J Control Release 1989; 9: 169–175.
11. Wilson HC, Cuff GW. Sustained release of isomazole from matrix tablets
administered to dogs. J Pharm Sci 1989; 78: 582–584.
M Pharm. (Pharmacognosy) 38
Chapter: 1.3 Introduction of Polymers
12. Dahl TC, Calderwood T, Bormeth A, et al. Influence of physicochemical
properties of hydroxypropyl methylcellulose on naproxen release from sustained
release matrix tablets. J Control Release 1990; 14: 1–10.
13. Banker G, Peck G, Williams E, et al. Microbiological considerations of polymer
solutions used in aqueous film coating. Drug Dev Ind Pharm 1982; 8: 41–51.
14. Anonymous. Final report on the safety assessment of hydroxyethylcellulose,
hydroxypropylcellulose, methylcellulose, hydroxypropyl methylcellulose and
cellulose gum. J Am Coll Toxicol 1986; 5(3): 1–60.
15. FAO/WHO. Evaluation of certain food additives and contaminants. Thirty-fifth
report of the joint FAO/WHO expert committee on food additives. World Health
Organ Tech Rep Ser 1990; No. 789.
16. Lewis RJ, ed. Sax’s Dangerous Properties of Industrial Materials, 11th edn. New
York: Wiley, 2004: 2054.
M Pharm. (Pharmacognosy) 39
Chapter: 1.4 Introduction of Mouth Ulcer
Mouth ulcers are open sores, which appear in the mouth. They are white or yellow in color, and are normally accompanied with a sharp pain, which is felt most when the person is eating. The pain is quite sharp when salty or spicy food passes over the ulcer. Mouth ulcers can occur wherever in the mouth - on the inner surface of the cheeks, lips, tongue, palate and at the base of the gums 1.
Figure 1.9 Mouth Ulcer
Symptoms 2
The symptoms preceding the ulcer may vary according to the cause of the ulcerative
process.
Some oral ulcers may begin with a sharp stinging or burning sensation at the site of
the future mouth ulcer. In a few days, they often progress to form a red spot or bump,
followed by an open ulcer. Sometimes this takes a little bit longer, depending on the
cause of the ulcer.
The oral ulcer appears as a white or yellow oval with an inflamed red border.
Sometimes a white circle or halo around the lesion can be observed. The grey, white,
or yellow coloured area within the red boundary is due to the formation of layers of
fibrin, a protein involved in the clotting of blood. The ulcer, which itself is often
extremely painful, especially when agitated, may be accompanied by a painful
swelling of the lymph nodes below the jaw, which can be mistaken for toothache.
Causes of mouth ulcers1
1. Incorrect diet
2. Digestive problems like constipation
3. Hormonal imbalances, particularly during puberty in girls
4. Anemia
5. Constant stress
6. Genetic factors
7. Illness of the herpes simplex virus
M.Pharm. (Pharmacognosy) 40
Chapter: 1.4 Introduction of Mouth Ulcer
8. Irritation by a few chemicals, tobacco and alcohol
In some cases there are infectious agents that are both bacterial and viral in nature that
are considered as causes of mouth ulcers. The various chemical compounds that are
found within the infectious agents are perhaps one of the reasons for mouth ulcers
forming 3.
Possible Complications 3
Cellulitis of the mouth, from secondary bacterial infection of ulcers
Dental infections (tooth abscesses)
Oral cancer
Spread of contagious disorders to other people
Treatment of mouth ulcers 2, 4
Treatments based on antibiotics and steroids are reserved for severe cases, and should
be used only under medical supervision.
Some doctors may also prescribe local anaesthetic, such as lidocaine, for cases of
multiple or severe oral ulcers.
Some people benefit from using the over-the-counter topical gel Bonjela, which
contains choline salicylate -- choline salicylate is a local analgesic that helps to reduce
the pain and inflammation associated with oral ulcers.
No single factor is solely responsible for the initiation of Aphthous lesion. In addition,
nutrient deficiencies need to be corrected and anti-inflammatory nutrients prescribed.
Herbs which are useful in the Treatment of Mouth Ulcers1
1. Licorice ( Glycyrrhiza glabra)
Licorice is used for a number of oral and dental problems. It is a piece of several
toothpaste brands. Its stem and leaves wash the mouth effectively. Apart from
controlling the sores, it can refresh the mouth and wash the teeth.
2. Kattha (Acacia catechu)
Kattha is an extract of the Acacia catechu plant, known normally in the west as the
catechu plant. This has caustic properties. It has a extraordinary place in Ayurvedic
medicine in oral treatment. It is used in many states or forms for the treatment of
ulcers.
M.Pharm. (Pharmacognosy) 41
Chapter: 1.4 Introduction of Mouth Ulcer
3.Banyan(Ficusreligiosa)
A decoction of the bark of the banyan tree decreases the pain caused by mouth ulcers.
4. Chebulic myrobalan (Tarminalia chebula)
The chebulic myrobalan is a component of the Triphala choorna, of which the amalaki
is also a necessary component. Its bark helps in decreasing the pain of the ulcers. It
also helps in correcting the constipation troubles, which causes ulcers.
5. Fenugreek (Trigonella foenum graecum)
Fenugreek leaves assists in the treatment of mouth ulcers. A mixture of these leaves is
used for gargling. Fenugreek is a strong mediator on the ulcers. Hence it is used as a
medicine for the recurrent ulcers.
6. Henna (Lawsonium alba)
Henna is a cooling herb and this herb can provide a soothing effect on the ulcers. It is
used with water for gargling to prevent mouth ulcers.
7. Indian Gooseberry (Emblica officinalis)
The Indian gooseberry or amalaki has a dual impact on ulcers. Used as a gargling
solution, it can loosen up the pain of the ulcers. Secondly it can deal with
constipation, which is usually one of the important factors causing ulcers in the
mouth.
8. Turmeric (Curcuma longa)
Turmeric is also a cooling agent as this helps in relieving mouth ulcers. It is mixed in
water and the suspension can be used for gargling for mouth ulcers treatment.
References :
1) http://www.himalayahomeremedies.com/homeremedies_mouthulcers.htm
2) http://www.medic8.com/healthguide/articles/mouthulcers.html
3) http://ulcertreatmentinfo4u.com/Causes-Of-Mouth-Ulcers.php
4) Joseph E.P., Michale T.M., Text Book of Natural Medicine, Vol-II, II nd edi.
1085.
M.Pharm. (Pharmacognosy) 42
Chapter: 2.1 Review of Literature on Glycyrrhiza glabra and Glycyrrhizin
Jeff burgess et al., reviewed Over-the-counter Treatments for Aphthous Ulceration
and Results from Use of a Dissolving Oral Patch Containing Glycyrrhiza Complex
Herbal Extract. The aim of this article was to present a review of over-the-counter
(OTC) treatment strategies used for aphthous ulcerations and to provide results from
the use of an herbal extract containing glycyrrhiza.
Haley, Jeffrey T have patented Licorice root extract oral patch for treating canker
sores. A method for treating mouth ulcers (canker sores/aphthous ulcers) with licorice
extracts oral patches to speed healing and relieve pain. If licorice extract is applied to
a mouth ulcer using an adhesive oral patch that delivers the medication for at least 30
minutes and the patches are used for at least two or more hours per day, the method
reduces the healing times from typically 10–14 days to typically 2 days. The licorice
extract patches also quickly reduces canker sore pain and, if used before commencing
a meal, reduces pain during the meal.
Deb. soumitra and mandal, Kumar S. have developed an estimation method of 18
B-glycyrrhetinic acid obtained from glycyrrhizin in Glycyrrhiza glabra (yastimadhu)
by Thin Layer Chromatography- Densitometric method. Beer’s law is obeyed in the
concentration range of 0.48 to 2.4 g/l. the recovery is 89-107%. The method is
simple and is applicable both for crude drug and polyherbal formulations containing
yastimadhu.
Chauhan S.K. et al., have developed and described a reverse phase high pressure
liquid chromatography method to determine glycyrrhizin in G. glabra and its extract.
The method involved separation of compound using the mobile phase Acetonitrile:
Water: Phosphoric acid (32:67:1) and detection of chromatogram at 250 nm using
photodiode array detector. The sensitivity of the method was observed to be 2.0 g
and the linearity was observed in the range of 2.0 g-16.0 g/l. The propose method
being precise, sensitive and reproducible, can be used for det5ection, monitoring and
quantification of glycyrrhizin from Glycyrrhiza glabra and its extract.
Chauhan S.K. et al., have developed and described a simple reproducible HPTLC
method for the determination of glycyrrhizin from Glycyrrhiza glabra and its
extract .The sensitivity was found to be linear in the range of 0.2 to 1.0 g. the
proposed method being precise, sensitive and reproducible can be used for detection,
monitoring and quantification of glycyrrhizin from Glycyrrhiza glabra and its
extract.
M. Pharm. (Pharmacognosy) 43
Chapter: 2.1 Review of Literature on Glycyrrhiza glabra and Glycyrrhizin
Racková et. Al., studied the Mechanism of anti-inflammatory action of liquorice
extract and glycyrrhizin. The antiradical activity, protective effect against lipid
peroxidation of liposomal membrane, and inhibitory effect on whole blood reactive
oxygen species (ROS) liberation of Glycyrrhiza glabra crude extract and glycyrrhizin,
its major compound, were assessed. The liquorice extract showed significant activity
in all the three assay systems used in a dose dependent manner. It displayed
remarkable reactivity with free stable 1,1'-diphenyl-2-picrylhydrazyl (DPPH) radical,
inhibitory efficacy in peroxidatively damaged unilamellar dioleoyl
phosphatidylcholine (DOPC) liposomes, and inhibition of ROS chemiluminescence,
generated by whole blood, induced by both receptor-bypassing stimuli (PMA) and
receptor operating stimuli (Opz) in the ranking order of stimuli PMA> Opz. These
activities may be attributed to phenolic antioxidants involving isoflavan derivatives,
coumarins and chalcones. Nonetheless, triterpene saponin glycyrrhizin exhibited no
efficacy in the system of DPPH reaction and peroxidation of liposomal membrane,
and negligible inhibition of chemiluminescence generated by inflammatory cells.
These results indicate that the mechanism of anti-inflammatory effect of glycyrrhizin
most probably does not involve ROS and this major constituent is not responsible for
the inhibition effects of liquorice extract on neutrophil functions.
Gupta V.K. et. al., investigated the antimicrobial potential of Glycyrrhiza glabra
roots. Antimycobacterial activity of Glycyrrhiza glabra was found at 500 μg/ml
concentration. Bioactivity guided phytochemical analysis identified glabridin as
potentially active against both Mycobacterium tuberculosis H37Ra and H37Rv strains at
29.16 μg/ml concentration. It exhibited antimicrobial activity against both Gram-
positive and Gram-negative bacteria. Our results indicate potential use of licorice as
antitubercular agent through systemic experiments and sophisticated anti-TB assay.
M. Pharm. (Pharmacognosy) 44
Chapter: 2.1 Review of Literature on Glycyrrhiza glabra and Glycyrrhizin
References:
1. Jeff burgess, Peter van der ven, Michael Martin, Jeffrey Sherman, The Journal of
contemporary dental practice, March 2008, 9(3).
2. Haley, Jeffrey T., Licorice root extract oral patch for treating canker sores , United
States Patent 7201930
3. Deb. Soumitra and Mandal, Kumar S., An estimation method of 18 B-
glycyrrhetinic acid obtained from glycyrrhizin in Glycyrrhiza glabra (yastimadhu)
by Thin Layer Chromatography- Densitometric method, Indian Drugs, November
1999, 36(11), 687-688.
4. Chauhan S.K. et.al., Estimation of glycyrrhizin from Glycyrrhiza glabra and its
extract by High pressure liquid chromatography, Indian drugs, Aug 1999,36(8),
521-524.
5. Chauhan S.K. et. al., Determination of glycyrrhizin from Glycyrrhiza glabra and
its extract by HPTLC, Indian journal of pharmaceutical science, July-Aug
1998,60(4),251-254.
6. Rackova, Lucia et al., Mechanism of anti-inflammatory action of liquorice extract
and glycyrrhizin, Natural Product Research, December 2007, 21(14), 1234-1241.
7. Gupta V. K. et al., Antimicrobial potential of Glycyrrhiza glabra roots, Journal of
Ethnopharmacology, March 2008, 116 (2), Pages 377-380.
M. Pharm. (Pharmacognosy) 45
Chapter: 2.2 Review of Literature on Acaia Catechu and Catechin
Singh K.N. and Lal B., noted Traditional Uses of Khair (Acacia catechu Willd.) by
Inhabitants of Shivalik Range in Western Himalaya. Katha or decoction of heartwood
is applied in mouth and on tongue to cure mouth ulcer. It is also applied externally on
ulcers, boils, skin eruptions and on gums as disinfectant.
Desai D.S. and Laddha K. S. have developed a stability indicating high performance
thin layer chromatographic (HPTLC) method for quantification of catechin. The
mobile phase was Chloroform: Ethyl acetate: Methanol: Toluene: Formic acid
(12:8:4:2:2). The calibration curve of catechin in methanol was linear in the
concentaration range of 800-2800 ng. The mean values of correlation coefficient,
slope and intercept were 0.9986 + 0.046, 477.363 + 3.961,266329 + 7.492. The limit
of detection of catechin was 25ng and limit of quantification was 85ng.no interference
was found from decomposition products. The percent recovery of catechin using
described procedure was 102.91 + 0.0123 from green tea and 98.805 +1.438 from
pale catechu. Concentration of catechin from green tea, black tea pale catechu and
black catechu was estimated.
Stuart (1979) has referred the use of the boiled and strained aqueous extract of the
heartwood as an astringent for inflamed conditions of the throat, gums, and mouth and
also externally for boils and chronic ulcers.
Sane R.T. et al., studied Spectrophotometric method for the determination of
cyanidalol (+ catechin) from pharmaceutical preparations. The methods use the
formation of coloured species of the drug with reagent like para-amino-phenol,
resorcinol, phosphomolybdic acid, folin and ciocalteau’s phenol and
parapenylediamine dihydrochloride (PPDA) in basic medium. The absorbance values
of the coloured species are measured at the wavelength of maximum absorption. The
methods are statistically validated and are found to be precise and accurate.
Sawant S.S. et al., studied the method of estimation of catechin from Acacia catechu
by HPTLC. This method could be used for routine analysis of catechin from acacia
catechu since it is relatively simple, sensitive and accurate.
Veluri R. et al., studied Phytotoxic and Antimicrobial Activities of Catechin
Derivatives. (±) - Catechin is a potent phytotoxin, with the phytotoxicity due entirely
to the (−)-catechin enantiomer. (+)-Catechin, but not the (−)-enantiomer, has
antibacterial and antifungal activities. Tetramethoxy, pentaacetoxy, and cyclic
derivatives of (±)-catechin retained phytotoxicity. The results indicate that antioxidant
properties of catechins are not a determining factor for phytotoxicity. A similar
M. Pharm. (Pharmacognosy) 46
Chapter: 2.2 Review of Literature on Acaia Catechu and Catechin
conclusion was reached for the antimicrobial properties. Centaurea maculosa (spotted
knapweed) exudes (±)-catechin from its roots, but the flavanol is not re-absorbed and
hence the weed is not affected. The much less polar tetramethoxy derivative may,
however, be absorbed and hence be able to cause toxicity. Because of the combination
of phytotoxicity and antimicrobial activity, (±)-catechin could be a useful natural
herbicide and antimicrobial.
Li ZhongXing et al., studied in-vitro study for anti-bacterial activity of Acacia
catechu (L.) Willd in 308 strains of clinical isolates 100 g of A. catechu was decocted
to make 200 ml of 50% catechu solution. Then different catechu concentrations were
made and used to study their antibacterial activity. Bacteria were isolated from
patients' blood, urine, abscesses and phlegm. These were Staphylococcus aureus (112
strains), Staphylococcus epidermidis (112 strains), Enterobacter aerogenes (28
strains), Klebsiella pneumoniae (28 strains) and Escherichia coli (28 strains). It was
found that the minimum inhibitory concentration (MIC) 50 values for catechu were
0.59, 1.19 and 1.19 mg/ml for S. aureus, S. epidermidis and E. aerogenes,
respectively, while the MIC 90 values were 1.19, 2.38 and 1.19 mg/ml. The MIC 90
value was 1.19 mg/ml for both K. pneumoniae and E. coli. It was concluded that
catechu has strong inhibitory activity on Gram +ve cocci and Gram -ve bacilli.
References:
1. Singh K.N. Singh and Lal B., Traditional Uses of Khair (Acacia catechu Willd.)
by Inhabitants of Shivalik Range in Western Himalaya, Ethnobotanical Leaflets
10, 2006, 109-112.
2. Desai D.S. and Laddha K. S., Stability indicating HPTLC determination and ph
stability profile of catechin, Indian Drugs, February 2002, 39(2), 91-95.
3. Stuart M., Reference section. In: Stuart M (Ed.), The Encyclopedia of Herbs and
Herbalism. London: Orbis Publishing, 1979 .141-283.
4. Sane R. T. et. al., Spectrophotometric methods for the determination of Cynidanol
(+ Catechin) from Pharmaceutical Preparations, Indian Drugs, 1984, 22(1), 20-24.
5. Sawant S. S. et. al., Estimation of Catechin from Acacia catechu by HPTLC,
Indian Drugs, 1995, 32(9), 461-463.
6. Veluri R. et al., Phytotoxic and antimicrobial activities of Catechin derivatives, J.
Agric. Food Chem., 2004, 52 (5), 1077–1082
7. Li ZhongXing, et al., Chinese Journal of Information on Traditional Chinese Med.
M. Pharm. (Pharmacognosy) 47
Chater:2.3 Review of Literature on Clove and Eugenol
Ayoola G. A. et al., studied Chemical analysis and antimicrobial activity of the
essential oil of Syzigium aromaticum (clove). The antimicrobial sensitivity of the
volatile oil against some Gram-negative bacteria (Escherichia coli ATCC 35218,
Escherichia coli, Klebsiella pneumoniae, Salmonella paratyphi, Citrobacter spp. and
Enterobacter cloacae), a Gram-positive bacterium (Staphylococcus aureus ATCC
25923), and a fungus (Candida albicans) showed a broad spectrum of activity.
Antioxidant screening of clove oil with 2,2-diphenylpicryl-hydrazyl radical (DPPH)
was positive, indicating the presence of free radical scavenging molecules which can
be attributed to the presence of eugenol, a phenolic compound.
Dr. Chetan has reviewed that the clove oil has antiseptic, antifungal, and anti-
bacterial properties and has been widely used to prevent oral diseases such as plaque
and mouth ulcers.
Dighe V.V et al. Have developed a simple , precise and quanititative HPTLC method
for determination of eugenol in Cinnamomum zeylanicum leaf powder. The
methanolic extract of cinnamomum zeylanicum were applied on TLC pre –coated
plate (merck) and was developed using Toluene: Ethyl acetate: Formic acid (9:1:0.1)
v/v/v as the mobile phase. Detection was carried out densitometrically using an UV
detector at 280 nm. The HPTLC proposed method is precise, accurate and rapid for
determination of eugenol.
Anandjiwala S. et al., have quantified 4 marker compounds, viz., eugenol, luteolin,
ursolic acid, and oleanolic acid, from the leaf of green and black varieties of O.
sanctum using high-performance thin-layer chromatography (HPTLC) with
densitometry. The methods were found to be precise, with relative standard
deviation (RSD) values for intraday analyses in the range of 0.52 to 0.91%, 0.77
to 1.29%, 0.11 to 0.16%, and 0.34 to 0.42% and for interday analyses in the
range of 0.73 to 0.96%, 1.02 to 2.08%, 0.11 to 0.12%, and 0.39 to 0.64% for
different concentrations of eugenol, luteolin, ursolic acid, and oleanolic acid,
respectively. Instrumental RSD values were 0.24, 0.39, 0.21, and 0.18% for
eugenol, luteolin, ursolic acid, and oleanolic acid, respectively. Accuracy of the
methods was checked by conducting a recovery study at 3 different levels for
the 4 compounds, and the average recoveries were found to be 99.73, 99.3,
100.58, and 100.57%, respectively. Eugenol content ranged from 0.175 to
0.362% (w/w) and luteolin from 0.019 to 0.046% (w/w) in the samples
M. Pharm. (Pharmacognosy) 48
Chater:2.3 Review of Literature on Clove and Eugenol
analyzed. The HPTLC-densitometry methods for the quantification of the 4
markers in O. sanctum leaf will have the applicability in quality control.
Dighe V. V. et al., developed A simple, rapid and precise reverse-phase high
performance liquid chromatographic method for the quantitative determination of
eugenol from the extract of dried powder of Cinnamomum tamala leaves and its
polyherbal formulation. Chromatographic analysis was carried out on Zorbax C18
column (150 mm · 4.6 mm, 5 lm) with a mobile phase of mixture of Water,
Acetonitrile and Methanol in the volume ratio of 50:40:10, at a flow rate of 1.0 ml/
min. Quantitation was performed using a UV-visible detector at 210 nm. Good
linearity was obtained over the ranges of 0.20–3.0l g /ml. for eugenol.
Pathak S. B. et al., have reported a simple TLC densitometric method for the
quantification of eugenol and gallic acid in clove. The method was validated for
precision, repeatability and accuracy. The contents of eugenol and gallic acid in
different samples of clove, as estimated by the proposed method, were found to be in
the range of 12.9–14.6% and 0.31–0.61% respectively. The proposed HPTLC method
for the estimation gallic acid and eugenol was found to be simple, precise, specific,
sensitive and accurate and can be used for routine quality control of clove.
Jyoti BB has reviewed Phytotherapeutics in conservative dentistry & endodontics. In
Conservative Dentistry & Endodontics, variety of dental materials & medicaments are
being used originated from plants. Identification & isolation of "Digoxin" from
Digitalis lanata, "Reserpine"from Rauwalfa serpentina, "Vincrystin "and "Vinblastin
"from Catharanthus rosea," Gutta percha "from Palaquiam species. "Eugenol "from
Svgygium aromaticum are some of the examples.
Lining Cai and Christine D. Wu studied the effect of crude MeOH extract of
Syzygium aromaticum (clove) exhibited preferential growth-inhibitory activity
against Gram-negative anaerobic periodontal oral pathogens, including
Porphyromonas gingivalis and Prevotella intermedia. By means of bioassay-directed
chromatographic fractionation, eight active compounds were isolated from this
extract and were identified as 5,7-dihydroxy-2-methylchromone 8-C-β-D-
glucopyranoside, biflorin, kaempferol, rhamnocitrin, myricetin, gallic acid, ellagic
acid, and oleanolic acid, based on spectroscopic evidence. The antibacterial activity
of these pure compounds was determined against Streptococcus mutans, Actinomyces
viscosus, P. gingivalis, and P. intermedia. The flavones, kaempferol and myricetin,
M. Pharm. (Pharmacognosy) 49
Chater:2.3 Review of Literature on Clove and Eugenol
demonstrated potent growth-inhibitory activity against the periodontal pathogens P.
gingivalis and P. intermedia.
Ogata M. et al., studied Antioxidant activity of Eugenol and related monomeric and
dimeric cmpounds. Since the inhibitory effect of eugenol (a), which was isolated as
an antioxidative component from plant, Caryopylli flos, on lipid peroxidation was
less than that of alpha-tocopherol, we synthesized the eugenol-related compounds
dieugenol (b), tetrahydrodieugenol (c), and dihydroeugenol (d), to find new strong
antioxidants and assessed them for their inhibitory effect on lipid peroxidation and
scavenging ability for superoxide and hydroxyl radicals. The antioxidative activities
were in the order: (b)>(c)> (d)> (a) for the thiobarbituric acid reactive substance
(TBARS) formation. These results suggest that the dimerized compounds have higher
antioxidant activities than that of the monomers. Electron spin resonance (ESR) spin
trapping experiments revealed that eugenol and its dimer, having allyl groups in the
structure, scavenged superoxide, and that only eugenol trapped hydroxyl radicals
under the conditions used. These finding suggest that eugenol and dieugenol have a
different mechanism of antioxidation, i.e. eugenol may inhibit lipid peroxidation at
the level of initiation, however , the related dimeric compounds may inhibit lipid
peroxidation at the level of propagation of free radical chain reaction like alpha-
tocopherol.
M. Pharm. (Pharmacognosy) 50
Chater:2.3 Review of Literature on Clove and Eugenol
References:
1. Ayoola G.A., Chemical analysis and antimicrobial activity of the essential oil of
Syzigium aromaticum (clove), African Journal of Microbiology Research., July
2008,2 , 162-166,
2. Dr. Chetan, Dentistry and Dental Information, 5 December, 2007.
3. Dighe V.V et al., Quantification of Eugenol from Cinnamomum zeylanicum
blume. leaf powder using High Performance Thin Layer Chromatography, Indian
drugs,43(6),June 2006,493-496.
4. Anandjiwala S . et al., Quantification of Eugenol, Luteolin, Ursolic acid, and
Oleanolic acid in black and green varieties of Ocimum sanctum Linn. Using High-
Performance Thin-Layer Chromatography, Journal of AOAC Int., Nov-Dec 2006,
89(6):1467-74.
5. Dighe V.V et al., Quantitative Determination of Eugenol from Cinnamomum
tamala Nees and Eberm. Leaf Powder and Polyherbal Formulation Using Reverse
Phase Liquid Chromatography, Chromatographia, May 2005, 61(9-10), 443-446.
6. Pathak S. B. et al., TLC Densitometric Method for the Quantification of Eugenol
and Gallic Acid in Clove, Chromatographia, 2004,60, 241-244.
7. Jyoti BB , Phytotherapeutics in conservative dentistry & endodontics -a review
(2005) Medknow Publications, United States
8. Lining Cai and Christine D. Wu, Compounds from Syzygium aromaticum
Possessing Inhibitory Activity against Oral Pathogens, J. Nat. Prod., 1996, 59
(10), 987-990.
9. Ogata M ., et al., Chem Pharm Bull (Tokyo). Oct 2000, 48(10), 1467-1469.
M. Pharm. (Pharmacognosy) 51
Chapter:2.4 Review of Literature on Buccoadhesive Drug Delivery
Mahdi A. B. et al., evaluated the efficacy of bioadhesive hydrogel patches, made of
a pharmaceutical grade cellulose derivative, in the control of pain and as an aid to
healing of aphthous ulceration. A significant reduction in stimulated pain was
recorded following application of the patches to the ulcers (P<0.01). The patches
were found to adhere longer to large ulcers in the early stages of ulceration, when they
achieved their maximum protective and pain-attenuating effects. The ulcer size was
recorded daily by the patient and patients claimed a reduction in healing time
following patch therapy.
Sang-Chul Shin et al., studied Mucoadhesive and Physicochemical Characterization
of Carbopol-Poloxamer Gels Containing Triamcinolone Acetonide. The viscosity
and bioadhesive property of Carbopol-Poloxamer gels containing triamcinolone
acetonide to mucosa were tested according to various concentrations of Carbopol
gels of various pH. The increase in Carbopol concentration caused increased
viscosity and bioadhesiveness. The neutralization of pH in various concentrations
of Carbopol gels showed the increased viscosity, showing the highest viscosity
and highest bioadhesiveness when neutralized to pH 6. According to FTIR and
XRD studies, the drug did not show any evidence of an interaction with the
polymers used and was present in an unchanged state.
Mizrahi B. et al., Studied the Mucoadhesive Polymers for Delivery of Drugs to the
Oral Cavity. Local therapy of the oral cavity is used to treat conditions such as
gingivitis, oral candidosis, oral lesions, dental caries, xerostoma and oral carcinomas.
Delivery systems used include mouthwashes, aerosol sprays, chewing gums,
bioadhesive tablets, films, gels and pastes. Prolonged contact time of a drug with body
tissue can significantly improve the clinical performance of many agents used for
treating oral disorders. These improvements range from better treatment of local
pathologies to improved drug bioavailability and controlled release to enhanced
patient compliance. There are abundant examples in the literature over the past 15
years of these improvements using bioadhesive polymers.
Chien, Yie, W.; NAIR, Mona; have patented the mucosal adhesive device for long-
acting delivery of pharmaceutical combinations in oral cavity. Mucosal
adhesive devices are provided for use in the oral cavity for therapy against
infections. The devices are dosage units which comprise a combination of
antimicrobial agents such as antifungal agents and anti-inflammatory agents,
M. Pharm. (Pharmacognosy) 52
Chapter:2.4 Review of Literature on Buccoadhesive Drug Delivery
optionally also a local anesthetic. The dosage units yield a gradual and
relatively constant release of the pharmaceuticals over at least a 12-hour period.
Salamat-Miller Nazila et al., have studied The use of mucoadhesive polymers in
buccal drug delivery. Buccal delivery of the desired drug using mucoadhesive
polymers has been the subject of interest since the early 1980s. Advantages associated
with buccal drug delivery have rendered this route of administration useful for a
variety of drugs. This review highlights the use of mucoadhesive polymers in buccal
drug delivery. Additionally, they focus on the new generation of mucoadhesive
polymers such as thiolated polymers, followed by the recent mucoadhesive
formulations for buccal drug delivery.
Ramana MV et.al., studied the Design and evaluation of mucoadhesive buccal drug
delivery systems containing Metoprolol tartrate. Mucoadhesive buccal tablets of
metoprolol tartarate were fabricated with objective of avoiding first pass metabolism
and prolonging duration of action. The mucoadhesive polymers used in formulation
were Carbopol-934, hydroxypropylmethylcellulose, hydroxyethylcellulose and
sodium carboxymethylcellulose. The formulations were characterized for
physiochemical parameters, in vitro release studies and in vivo placebo studies. The
best mucoadhesive performance and in vitro drug release profile were exhibited by
the tablets containing hydroxyethylcellulose and Carbopol-934 in ratio 1:2.
Khanna R. et al., have prepared and evaluated the mucoadhesive buccal films of
clotrimazole for oral candida infections. Mucoadhesive buccal films of clotrimazole
for local delivery of the drug to the oral cavity were formulated by the solvent
casting technique. A number of different bioadhesive and film- forming polymers
were evaluated. Propylene glycol was used as the plasticizer while the solvents
depended on the type of polymer chosen. The films were evaluated on the basis of
their physical characteristics, biadhesive performance, release characteristics,
surface ph, folding endurance and strechability. A combination of carbopol-934P
and hydroxy propyl cellulose- M in the ratio of 1:5 and using ethanol (95%) as the
solvent was found to give satisfactory results. The film exhibited an in vitro
adhesion time of 4 hours and maintained the concentration of clotrimazole in the
dissolution medium(isotonic phosphate buffer ph 6.6) above the MIC of candida
albicans for upto 4 hours. A maximum concentration of 21.1 µg/ml was obtained in
the dissolution medium after 2 hours. The drug release from the formulation was
found to be microbiologocally active.
M. Pharm. (Pharmacognosy) 53
Chapter:2.4 Review of Literature on Buccoadhesive Drug Delivery
Mizrahi B.et al., developed a bioadhesive erodable patch for treating mouth ulcers.
Adhesive patches were prepared by compression molding of mixed polymeric
powders, absorbed with citrus oil and magnesium salt .A clinical trial was conducted
on 248 volunteers suffering from mouth ulcers. Patches gradually erode for eight
hours releasing the citrus oil in a zero order pattern while the magnesium is released
during a period of two hours. Patches were effective in reducing pain and decreasing
healing time (p<0.05) without adverse side effects . The mucoadhesive patch was
found to be highly effective for pain reduction and healing time in both single ulcer
and recurrent aphthous stomatitis patients.
Kim TH et al., have prepared and Characterized the novel mucoadhesive polymer
blend film consisting of Carbopol, poloxamer, and hydroxypropylmethylcellulose
(HPMC). Triamcinolone acetonide (TAA) was loaded into
Carbopol/poloxamer/HPMC polymer blend film. Swelling ratio of
Carbopol/poloxamer/HPMC films was lowest in Carbopoll poloxamer/HPMC at
mixing ratio of 35/30/35 (wt/wt/wt). Adhesive force of Carbopol/poloxamer/HPMC
films increased with increasing HPMC content in Carbopol/poloxamer/HPMC
polymer blend film and increasing hydroxypropyl group content in HPMC due to
hydrophobic property of HPMC although bioadhesive force was highest at mixing
ratio of 35/30/35 (wt/wt/ wt). Release of TAA from TAA-loaded
Carbopol/poloxamer/HPMC polymer blend film in vitro increased with increasing
loading content of drug.
Dhiman M. et al., prepared and evaluated in vitro the bioadhesive gels of 5-
Fluorouracil (FU) for the treatment of oropharyngeal cancer. The gel formulations
containing FU were prepared by using Poloxamer 407, HPMC K 15 M, and Gantrez
S-97 (polymethylvinylether-co-maleic anhydride). The formulations contained
Poloxamer 407 (16-18% w/w) either alone or in combination with HPMC K 15 M and
Gantrez S-97. The bioadhesiveness of the gels was found to increase with increasing
proportion of HPMC K 15 M and Gantrez S-97. In vitro release studies indicated that
release could be sustained up to 8 hr. Increasing temperature increased the drug
release by increasing drug diffusion despite increase in viscosity. The pH of the
release medium showed a very slight effect on the release of FU.
Ali J. et al., designed and characterized Buccoadhesive erodible disk for treatment of
oro-dental infections. The optimized disk containing 5.0 mg of cetylpyridinium
chloride, 2.0 mg of magnesium stearate and 6.0 mg of mannitol along with sodium
M. Pharm. (Pharmacognosy) 54
Chapter:2.4 Review of Literature on Buccoadhesive Drug Delivery
carboxy methyl cellulose DVP and hydroxypropylmethylcellulose K4M in the ratio of
1:3 was found to release the drug for a period of over 6.0 h without getting dislodged.
Maximum in vitro drug release was found to be 94.78% in 6.0-h study. The
bioadhesive performance and the surface pH of the disks were satisfactory.
Cetylpyridinium chloride disks were tested against microorganisms commonly found
in oro-dental infections namely Candida albicans, Staphylococcus aureus,
Escherichia coli and Streptococcus mutans. The disk as well as the in situ samples
showed inhibition of growth of microorganisms.
Varshosaz J. et al., developed a localized drug delivery system that offers prolonged
administration of metronidazole into the periodontal pocket, muccoadhesive gel
formulations containing 5% w/w metronidazole were prepared using the bioadhesive
polymers: carboxymethylcellulose, methylcellulose, hydroxyethylcellulose,
polyvinylpirrolidone, and carbopol. Increased concentrations of the polymers
decreased the drug release rate and enhanced syringeability, yield value, and
adhesiveness but decreased the spreadability. The bioadhesive properties of the gels
were affected by pH and Ca2+ concentration. The gel containing 20%
hydroxyethylcellulose, 20% polyvinylpirrolidone, and 1% carbopol exhibited zero-
order drug release kinetics and suitable physical properties for drug delivery to the
periodontal pocket.
M. Pharm. (Pharmacognosy) 55
Chapter:2.4 Review of Literature on Buccoadhesive Drug Delivery
References:
1. Mahdi AB, Coulter WA, Woolfson AD, Lamey PJ, Journal of oral pathology
& medicine, 1996 Sep;25(8):416-9
2. Sang-Chul Shin ; Ja-Young Kim ; In-Joon Oh, Drug development and industrial
pharmacy, 2000 Mar;26(3):307-12
3. Mizrahi, Boaz; Domb, Abraham J., Recent Patents on Drug Delivery &
Formulation, Volume 2, Number 2, June 2008 , pp. 108-119(12)
4. Chien, Yie, W.; Nair, Mona, Mucosal adhesive device for long-acting delivery of
pharmaceutical combinations in oral cavity, Patent no.WO/1995/015137
5. Salamat-Miller Nazila ; Chittchang Montakam ; Johnston Thomas P. , Advanced
drug delivery reviews , 2005, vol. 57, no 11 (171 p.)
6. M.V.Raman, C Nagda, M Himaja, Indian Journal of Pharmaceutical Science, Year
: 2007 , Volume : 69 , Issue : 4 , Page : 515-518
7. R.Khanna, S.P.Agarwal, Alka Ahuja, Indian Journal Of Pharmaceutical
Science,1997,59(6),page.299-305
8. http://www.cankercover.comram
9. Kim TH , Ahn JS, Choi HK, Choi YJ, Cho CS. Arch Pharm Res. 2007
Mar;30(3):381-6
10. Dhiman M , Yedurkar P, Sawant KK. Pharmaceutical development and
technology, 2008; 13(1):15-25
11. J. Ali, R. Khar, A. Ahuja and R. Kalra, International Journal of Pharmaceutics,
Volume 238, Issues 1-2, 15 May 2002, Pages 93-103
12. Jaleh Varshosaz ; Nasser Tavakoli ; Sharona Saidian , Drug Delivery, Volume 9,
Issue 2 April 2002 , pages 127 - 133
M. Pharm. (Pharmacognosy) 56
Chapter: 2.5 Review of Literature on Mouth Ulcer
Moghadamnia A. A. et. al. studied the efficacy of the bioadhesive patches containing
licorice extract in the management of recurrent aphthous stomatitis. This study
evaluated the efficacy of licorice bioadhesive hydrogel patches to control the pain and
reduce the healing time of recurrent aphthous ulcer. According to the results of this
study, licorice bioadhesive can be effective in the reduction of pain and of the
inflammatory halo and necrotic center of aphthous ulcers.
Stuart (1979) has referred the use of the boiled and strained aqueous extract of the
heartwood as an astringent for inflamed conditions of the throat, gums, and mouth and
also externally for boils and chronic ulcers.
Mahdi A. B. et al., evaluated the efficacy of bioadhesive hydrogel patches, made of a
pharmaceutical grade cellulose derivative, in the control of pain and as an aid to
healing of aphthous ulceration. A significant reduction in stimulated pain was
recorded following application of the patches to the ulcers (P<0.01). The patches
were found to adhere longer to large ulcers in the early stages of ulceration, when they
achieved their maximum protective and pain-attenuating effects. The ulcer size was
recorded daily by the patient and patients claimed a reduction in healing time
following patch therapy.
Mizrahi B. et al., developed a bioadhesive erodable patch for treating mouth ulcers.
Adhesive patches were prepared by compression molding of mixed polymeric
powders, absorbed with citrus oil and magnesium salt. A clinical trial was conducted
on 248 volunteers suffering from mouth ulcers. Patches gradually erode for eight
hours releasing the citrus oil in a zero order pattern while the magnesium is released
during a period of two hours. Patches were effective in reducing pain and decreasing
healing time (p<0.05) without adverse side effects. The mucoadhesive patch was
found to be highly effective for pain reduction and healing time in both single ulcer
and recurrent aphthous stomatitis patients.
Hau; Kee Hung patented the invention that provides a medicament for topically
treating acute bacterial infections in the oral mucosa, and methods of use. The
medicament comprises a dry dosage (such as a troche or powder) of one or more
antibacterial agents and, preferably, one or more polyvalent metal compounds. The
medicament is directly applied to the site of the infection and dissolves in saliva,
within about 5 to about 15 minutes, thereby directly delivering a supratherapeutic
dosage of the antibacterial agent to the infected oral tissue. Further, in a preferred
embodiment the medicament directly delivers a therapeutically high concentration of
M. Pharm. (Pharmacognosy) 57
Chapter: 2.5 Review of Literature on Mouth Ulcer
a polyvalent metal compound in suspension to the infected area, thereby forming a
protective barrier over the infected oral tissue.
Campbell, Phillip patented Lesion and ulcer medication .The invention is based on
the discovery that certain combinations of antimicrobial agents, anti-inflammatories,
mid antihistamines, along with an accompanying mucoadhesive provide unexpected
and highly effective intraoral ulcer medications. The present invention provides an
intraoral ulcer medication which prevents both secondary infection and promotes
healing while simultaneously providing immediate relief from pain. This ulcer
medication comprises an antimicrobial, an anti-inflammatory, an antihistamine, and
optional antifungal and anesthetic components along with a mucoadhesive or
equivalent.
References:
1. Moghadamnia A. A. , Motallebnejad M. , Khanian M., The efficacy of the
bioadhesive patches containing licorice extract in the management of recurrent
aphthous stomatitis, Phytotherapy Research,2008, 23(2), 246 – 250
2. Stuart M, Reference section. In: Stuart M (Ed.) The Encyclopedia of Herbs and
Herbalism. 1979 pp. 141-283. London: Orbis Publishing
3. Mahdi AB, Coulter WA, Woolfson AD, Lamey PJ, Journal of oral pathology
& medicine, 1996 Sep;25(8):416-9
4. http://www.cankercover.comram
5. Hau; Kee Hung, Lesion-directed dry dosage forms of antibacterial agents for the
treatment of acute mucosal infections of the oral cavity, United States Patent
6,248,718, June 19, 2001.
6. Campbell, Phillip, Lesion and ulcer medication, US Patent 6352711
M. Pharm. (Pharmacognosy) 58
Chapter: 3 Aim and Plan of the Work
Aim of the work:
The prime aim of this investigation is to develop the Gel and Patch formulations of
the Glycyrrhiza glabra extract, Acacia catechu extract and clove oil for the mouth
ulcer and to evaluate them for in vitro release efficiency. These formulations are also
evaluated for various formulation parameters.
Plan of the work:
The plan of the work is divided in the following parts.
Procurement of Raw materials
Evaluation of Raw materials
Preparation of Extracts
Standardization of extracts by HPTLC
Preparation and Evaluation of Gel Formulations
Preparation and Evaluation of Patch Formulations
The evaluation of developed formulation is carried out as:
Evaluation of various physicochemical parameters of the gel like pH,
Viscosity, Spreadibility and Drug content.
Evaluation of various physicochemical parameters of the patch like Folding
endurance, Surface pH, In vitro Residence time and Drug content.
In vitro Drug Release study using cellophane membrane in modified diffusion
cell.
Antimicrobial Activity of Selected Formulations.
M. Pharm. (Pharmacognosy) 59
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 60
4.1 Experimental:
4.1.1 Procurement of Plant Materials
The whole material of Glycyrrhiza glabra (G. glabra) and Acacia catechu (A.
catechu) was procured from LVG and SONS and powder was made in laboratory and
passed through sieve no. 60. The clove oil was procured from Saraiya Chemicals,
Ahmedabad. The standard compounds, 18-β-glycyrhhetinic acid and Catechin was
procured from Sigma Aldrich Ltd., Banglore.
4.1.2 Evaluation of G.glabra whole Material and Powder
(1) Macroscopic study of Whole material and Powder
The morphology of G. glabra root was examined and photographed using camera.
The powder of G. glabra was tested for color, odor and taste. The preliminary
evaluation for foreign matter was also done to remove the admixture material if
present in sample may be as adulterant or substitute by visualization method.
(2) Microscopy study 1
The stained and unstained slide of G. glabra was prepared and the characters were
examined photographed using CCD camera.
(3) Chemical Identification1
Test for Saponins and Flavanoids:
Foam test:
Mixed the powder with water. After shaking stable foam was produced due to
presence of saponins.
Liebermann-Burchard test:
Mixed 2 ml of extract with chloroform,1-2 drops of acetic anhydride and 2 drops of
conc. H2SO4 was added. Red color was produced due to the presence of triterpenois.
The drug powder with 80% v/v sulphuric acid shows orange yellow color due to
presence of flavanoids.
(4) Identification by TLC 2
TLC of G. glabra was carried out in the following conditions.
Stationary phase : Aluminium-backed silica gel 60F254 plate (E. Merck)
Mobile phase : Ethyl Acetate: Ethanol: Water: Ammonia (65:25:9:1)
Chamber saturation : 30 minutes
Standard solution : 18-β-glycyrhhetinic acid dissolved in methanol
Test sample : Dry aqueous extract of G. glabra was dissolve in methanol
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 61
Detection : (1) In U.V.254 nm
(2) After spraying with Anisaldehyde sulphuric acid
Sample ID : A. G. glabra extract
B. 18-β-glycyrhhetinic acid
(5) Physicochemical parameters
To ensure good quality of G. glabra powder, various physicochemical parameters was
performed as per WHO guidelines 5.
a) LOD at 105° C
b) Ash value
Total ash
Acid insoluble ash
Water soluble ash
c) Extractive value
Alcohol soluble extractive value
Water soluble extractive value
(6) Estimation of 18-β-glycyrhhetinic acid (18-β-G.A.) in G. glabra powder by
HPTLC
Preparation of std. stock solution: 100 µg per ml stock solution was prepared by
dissolving 1 mg 18-β-glycyrhhetinic acid in10 ml chloroform
Preparation of test solution: 1 gm of G. glabra powder was mixed with 50 ml of water
and extracted with 25 ml chloroform. The chloroform was evaporated and residue was
dissolved in 10 ml chloroform.
Test solution of 3 µl was applied along with 3, 4, 5, 6, 7 and 8 µl standard 18-β-
glycyrhhetinic acid solution on Aluminium- backed silica gel 60F254 plate (E.
Merck) (10 x 10 cm). It was followed by development of plate up to 85 mm at 25º C
and the chomatogram was recorded by scanning at 254 nm on a CAMAG TLC
scanner. The Amount of 18-β-glycyrhhetinic acid was determined using the
calibration curve plotted between concentration and peak area of standard 18-β-
glycyrhhetinic acid.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 62
Chromatogrhaphic condition:
Stationary phase : 10 x 10 cm Aluminium-backed silica gel 60F254 plate
(E.Merck)
Mobile phase : Ethyl Acetate: Ethanol: Water: Ammonia (65:25:9:1)
Chamber saturation : 30 minutes
Band with : 5 mm
Distance of band : 7 mm
Rate of spotting : 10 sec/ µl
Distance run : 85 mm
Scanning wavelength : 254 nm
Scanning speed : 5mm/ sec
Slit dimension : 5.0 x 0.45 mm
Temperature : 25º C
4.1.3 Evaluation of A. catechu whole material and Powder
(1) Macroscopic study of Whole material and Powder
The morphology of A. catechu root was examined and photographed using camera.
The powder of A. catechu was tested for color, odor and taste. The preliminary
evaluation for foreign matter was also done to remove the admixture material if
present in sample may be as adulterant or substitute by visualization method.
(2) Chemical Identification 1
Test for Phenolic compounds and tannins
Ferric chloride test:
To 1 ml of extract, 5% ferric chloride solution added, formation of dark blue or
greenish black color shows the presence of the tannins.
Lead acetate test:
The test solution mixed with lead acetate solution. Formation of white precipitates
indicates the presence of tannins.
Match stick test:
A matchstick was dipped in decoction of black catechu, dried in air and dipped it in
concentrated hydrochloric acid and warmed it near the burner. Magenta or purple
color was produced.
Vanillin –Hydrochloric acid test:
(Vanillin 1: Alcohol 10: dil. hydrochloric acid 10) catechu showed pink or red color.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 63
(3) Identification by TLC 3
TLC of A. catechu was carried out in the following conditions.
Stationary phase : Aluminium-backed silica gel 60F254 plate (E. Merck)
Mobile phase : Toluene: Ethyl Acetate: Methanol: Glacial Acetic Acid
(2.7:6:1:0.3)
Chamber saturation : 30 minutes
Test sample : Dry aqueous extract of acacia catechu was dissolved in
methanol.
Detection : (1) In U.V.254 nm
(2) After spraying with vanillin sulphuric acid
(4) Physicochemical parameters
To ensure good quality of A. catechu powder, various physicochemical parameters
was performed as per WHO guidelines 5.
d) LOD at 105° C
e) Ash value
Total ash
Acid insoluble ash
Water soluble ash
f) Extractive value
Alcohol soluble extractive value
Water soluble extractive value
(5) Estimation of Catechin in A. catechu powder by HPTLC
Preparation of std. stock solution : 100 µg per ml stock solution was prepared by
dissolving 1 mg catechin in10 ml methanol
Preparation of test solution: 1 gm of acacia catechu powder was mixed with 20 ml of
water, filtered and evaporated to dryness. The residue was dissolved in 50 ml
methanol and filtered. From this 1 ml of solution was taken and diluted up to 10 ml
with methanol. From this 1 ml of solution taken and mixed with 1 ml of methanol.
Test solution of 6 µl was applied along with 5, 6, 7, 8 and 9 µl standard Catechin
solutions on a Aluminium- backed silica gel 60F254 plate (E. Merck) (10 x 10 cm).
Then the plate was developed up to 85 mm at 25º C and the chomatogram was
recorded by scanning at 254 nm on a CAMAG TLC scanner. The Amount of catechin
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 64
was determined using the calibration curve plotted between concentration and peak
area of standard catechin.
Chromatogrhaphic condition:
Stationary phase : 10 x 10 cm Aluminium-backed silica gel 60F254 plate
(E.Merck)
Mobile phase : Toluene: Ethyl Acetate: Methanol: Glacial Acetic Acid
(2.7:6:1:0.3)
Chamber saturation : 30 minutes
Band with : 5 mm
Distance of band : 7 mm
Rate of spotting : 10 sec/ µl
Distance run : 85 mm
Scanning wavelength : 254 nm
Scanning speed : 5mm/ sec
Slit dimension : 5.0 x 0.45 mm
Temperature : 25º C
(6) Estimation of total tannins in Acacia catechu 4
1 gm of Acacia caetchu powder was accurately weighed and introduced into a 250 ml
glass stopered flask, 100 ml of water was added and shaken for 1.0 hr. and kept
overnight. The material was allowed to settle and the liquid was filtered through a
filter paper, discarded first 20 ml of filtrate.10 ml of filtrate was transferred to a 1 liter
conical flask. To this filtrate 750 ml water and 25 ml of Indigosulphonic acid solution
were added. This solution was titrated with 0.1 N Potassium Permanganate and
shaken vigorously till a golden yellow end point (T2) was reached. A blank
determination (T1) was also performed.
Each ml of 0.1 N Potassium Permanganate is equivalent to 0.004157 g of total
tannins.
Quantity of total tannins (%) = [(T2-T1) x actual normality x 0.004157 x1000]
W x 0.1
Where W = the total weight of the plant material
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 65
4.1.4 Evaluation of Clove oil
(1) Description
The clove oil was tested for its appearance, color, odor and taste.
(2) Determination of Refractive Index 6
The refractive index of clove oil was measured using Cyber Abbe Refractometer at
25º C (±0.5) with reference to the wavelength of the D line of sodium (Ψ =589.3 nm).
The temperature was carefully adjusted and maintained since the refractive index
varies significantly with temperature. To achieve accuracy, the apparatus was
calibrated against distilled water: which has a refractive index of 1.3325 at 25 º C.
(3) Determination of Specific Gravity 6
Specific gravity of clove oil was determined using specific gravity bottle. The specific
gravity bottle was calibrated by filling it with recently boiled and cooled water at
25°C and weighing the contents. It was assumed that the weight of 1 ml of water at
25°C when weighed in air of density 0.0012 gm per ml is 0.99602 gm; calculate the
capacity of the specific gravity bottle. The temperature of clove oil was adjusted to
about 20° C and fills the specific gravity bottle with it and weighed. The tare weight
of the specific gravity bottle was substracted from the filled weight of specific gravity
bottle. The specific gravity was determined by dividing the weight in air, in g, of the
quantity of liquid which fills the specific gravity bottle at the specified temperature,
by the capacity expressed in ml, of the specific gravity bottle at the same temperature.
(4) Identification by TLC 2
TLC of clove oil was carried out in the following conditions.
Stationary phase : Aluminium- backed silica gel 60F254 plate (E. Merck)
Mobile phase : Toluene: Ethyl Acetate (93:7)
Chamber saturation : 30 minutes
Standard solution : Eugenol diluted with toluene
Test sample : Clove oil diluted with toluene
Detection : (1) In U.V.254 nm
(2) After spraying with Anisaldehyde sulphuric acid
Sample ID : A. Diluted Clove Oil
B. Eugenol
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 66
(5) Estimation of Eugenol in clove oil by HPTLC
Preparation of std. stock solution: 50 µl eugenol was diluted up to 50 ml with
methanol. From this 4 ml of solution was taken and diluted up to 50 ml with
methanol. Preparation of test solution: 100 µl of clove oil was diluted up to 10 ml
with methanol.
Test solution of 1.5 µl was applied along with 2.5, 5, 7.5,10 and12.5 µl standard
eugenol solution on a precoated silica gel 60 plate (10 x 10 cm). After that the plate
was developed up to 85 mm at 25º C and the chomatogram was recorded by scanning
at 254 nm on a CAMAG TLC scanner. The Amount of eugenol was determined using
the calibration curve plotted between concentration and peak area of standard
eugenol.
Chromatogrhaphic condition:
Stationary phase : 10 x 10 cm Aluminium-backed silica gel 60F254 plate
(E.Merck)
Mobile phase : Toluene: Ethyl Acetate (93:7)
Chamber saturation : 30 minutes
Band with : 5 mm
Distance of band : 7 mm
Rate of spotting : 10 sec/ µl
Distance run : 85 mm
Scanning wavelength : 254 nm
Scanning speed : 5mm/ sec
Slit dimension : 5.0 x 0.45 mm
Temperature : 25º C
4.1.5 Preparation of extracts
Preparation of aqueous extracts of G. glabra powder and A. catechu powder
100 gm of G. glabra and A. catechu powder was extracted with 500 ml of water by
cold maceration for 24 hrs. The solution was filtered and evaporated to dryness in
Rotary evaporator at 60 ºC. The resulting extract was dried in dessicator.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 67
4.1.6 Standardization of extracts by HPTLC
(1) Standardization of extracts of G. glabra
Preparation of std. stock solution: 100 µg per ml stock solution was prepared by
dissolving 1 mg 18-β-glycyrhhetinic acid in10 ml chloroform
Preparation of test solution: 10 mg of aqueous extract of G. glabra was dissolved in
50 ml of chloroform.
Test solution of 7 µl was applied along with 3, 4, 5, 6, 7 and 8 µl standard 18-β-
glycyrhhetinic acid solution on Aluminium-backed silica gel 60F254 plate (E. Merck).
(10 x 10 cm). It was followed by development of plate up to 85 mm at 25º C and the
chomatogram was recorded by scanning at 254 nm on a CAMAG TLC scanner as per
the chromatographic condition mentioned in section 4.1.2.(6). The Amount of 18-β-
glycyrhhetinic acid was determined using the calibration curve plotted between
concentration and peak area of standard 18-β-glycyrhhetinic acid.
(2) Standardization of extracts of A. catechu
Preparation of std. stock solution: 100 µg per ml stock solution was prepared by
dissolving 1 mg 18-β-glycyrhhetinic acid in10 ml methanol.
Preparation of test solution: 10 mg of aqueous extract of Acacia catechu was
dissolved in 50 ml of methanol.
Test solution of 14 µl was applied along with 5, 6, 7, 8 and 9 µl standard Catechin
solution on an Aluminium-backed silica gel 60F254 plate (E.Merck) (10 x 10 cm).
After that the plate was developed up to 85 mm at 25º C and the chomatogram was
recorded by scanning at 254 nm on a CAMAG TLC scanner as per the
chromatographic condition mentioned in section 4.1.3.(5). The Amount of catechin
was determined using the calibration curve plotted between concentration and peak
area of standard catechin.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 68
4.2 Results and Discussion:
The result of various standardization parameters performed for ensuring the quality of
samples collected from market was as noted below
4.2.1 Evaluation of G.glabra whole Material and Powder
(1) Macroscopic study of Whole material and Powder
Figure: 4.1 Morphology of Glycyrrhiza glabra
The morphology of the sample was showed that outer surface was yellowish brown or
dark brown in colour, externally longitudinally wrinkled with patches of cork. fracture
was coarsely fibrous in bark and splintery in wood. This morphological characters
was giving identity of G. glabra.
The G. glabra powder having pale yellow to brown color with characteristic odor and
sweet taste. The foreign matter determination by visualization method was showing
absence of any admixture or any other obnoxious materials. The particle size of the
samples was characterized to 60 # by sieve analysis.
(2) Microscopy study 1
Cork cells Crystal fibres
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 69
Vessel Ca-oxalate crystal
Figure: 4.2 Microscopic characters of G. glabra powder sample
Microscopic characterization showed the presence of calcium oxalate crystals, cork
cells, crystal fibres and vessels when observed under stained and unstained slide. The
presence of this well defined diagnostic characters was giving identity of G. glabra
species.
(3) Chemical Identification
G. glabra extract shows positive result for Foam test, Liebermann-burchard test and
with 80% v/v sulphuric acid. So it contain triterpenoidal saponins and flavanoid
glycosides.
(4) Identification by TLC
In the TLC identification , violet spot was observed at Rf = 0.46 in extract of G.
glabra and in std. 18-β-glycyrhhetinic acid solution in U.V 254 nm and after spraying
with Anisaldehyde sulphuric acid reagent which confirm the identity of the G. glabra.
A B A B
(1) (2)
A: Std. 18-β-glycyrhhetinic acid, B: test solution of G. glabra powder extract ,
(1): In U.V. 254 nm , (2): After spraying with Anisaldehyde sulphuric acid
Figure: 4.3 TLC profile of Glycyrrhiza glabra powder
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 70
(5) Physicochemical parameters
The various physicochemical parameters performed for G. glabra was mentioned in
Table 4.1.
Table: 4.1 Physicochemical parameters of G. glabra powder
Sr. no. Physicochemical
parameters
Observation* Reference value
1 LOD at 105 ºC 5.3 ± 0.4 NMT 8 %
2 Ash value
A. Total ash 7.22 ± 0.09 NMT 10 %
B. Acid insoluble ash 2.21 ± 0.26 NMT 2.5%
C. Water soluble ash 0.77 ± 0.025 -
3 Extractive value
A. Alcohol extractive value 11.4 ± 0.52 NLT 10 %
B. Water extractive value 22.43 ± 0.51 NLT 20 %
* = Mean ± Standard Error Mean ( n = 3)
The G. glabra powder shown loss on drying within the pharmacopoeial limit. The
results of ash values were also within the pharmacopoeial limit as mentioned in the
table. The results of water soluble extractives and alcohol soluble extractives of
samples are mentioned in the table. High values in cases of of water soluble
extractives indicate the presence of good amount of water soluble chemical
constituents in the drug.
(6) Estimation of 18-β-glycyrhhetinic acid in G. glabra powder by HPTLC
The quantification of of 18-β-glycyrhhetinic acid in G. glabra powder was carried
out by HPTLC as per the procedure mentioned in Section 4.1.2 (6).
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 71
1 2 3 4 5 6 T 1 2 3 4 5 6 T
A B
A: In U.V. 254 nm, B: After spraying with Anisaldehyde sulphuric acid reagent
Figure: 4.4 HPTLC profile of G. glabra powder
Figure: 4.5 HPTLC chromatogram of standard 18-β-G.A.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 72
Figure: 4.6 HPTLC chromatogram of G. glabra extract
Table: 4.2 Observation table for calibration curve of std. 18-β- G.A.
(Using HPTLC method)
Standard solution Test
solution
Spot no. 1 2 3 4 5 6
µl of spot 3 4 5 6 7 8 3
Concentration
(mcg/µl)
0.3 0.4 0.5 0.6 0.7 0.8 -
Peak area 2172.2 2700 3197.7 3570.5 3766.4 4195.5 3178.9
% 18-β-G.A. - - - - - - 0.18 %
y = 3975.x + 1073.1R² = 0.977
0
1000
2000
3000
4000
5000
0 0.2 0.4 0.6 0.8 1
AU
C
Concentration (mcg/µl )
Figure: 4.7 Calibration curve of 18-β- G.A. by HPTLC method
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 73
The concentration of 18-β-glycyrhhetinic acid in G. glabra extract was calculated
using regression equation obtained from the standard curve which was found to be
0.18 %.
4.2.2 Evaluation of A. catechu whole material and Powder
(1) Macroscopic study of Whole material and Powder
Figure: 4.8 Morphology of A. catechu
The morphology of the sample was showed that it occurs as square blocks or irregular
pecies black to blackish brown in color. Outer surface was hard and brittle; when
broken fractured surface showed small cavities and soft. This morphological
characters was giving identity of A. catechu.
The A. catechu powder having brownish black color with astringent taste.
The foreign matter determination by visualization method was showing absence of
any admixture or any other obnoxious materials. The particle size of the samples was
characterized to 60 # by sieve analysis.
(2) Chemical Identification
Acacia catechu extract shows positive result for Match stick test, Vanilin
Hydrochloric acid test, 5% FeCl3 test and lead acetate test. So, it contain catechin and
tannins.
(3) Identification by TLC
In the TLC of A. catechu extract, grey spot at Rf = 0.6 in U.V. 254 nm and after
spraying with vanillin sulphuric acid reagent respectively in both extract and std.
solution.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 74
A B A B
(1) (2)
A: Std. catechin, B: Test solution of A. catechu powder extract ,
(1): In U.V. 254 nm, (2): After spraying with vanillin sulphuric acid reagent
Figure: 4.9 TLC profile of A. catechu powder
(4) Physicochemical parameters
The various physicochemical parameters performed for A. catechu was mentioned in
Table 4.3.
Table: 4.3 Physicochemical parameters of A. catechu powder
Sr. no. Physicochemical
parameters
Observation* Reference value
1 LOD at 105 ºC 4.38 ± 0.46 -
2 Ash value
A. Total ash 11.13 ± 0.32 NMT 15%
B. Acid insoluble ash 0.6 ± 0.26 NMT 2%
C. Water soluble ash 1.4 ± 0.46 -
3 Extractive value
A. Alcohol extractive value 43.53 ± 0.53 NLT 40 %
B. Water extractive value 50.5 ± 0.71 NLT 50 %
* = Mean ± Standard Error Mean ( n = 3)
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 75
The Acacia caetchu powder shown loss on drying within the pharmacopoeial limit.
The results of ash values, water soluble extractives and alcohol soluble extractives
were also within the pharmacopoeial limit as mentioned in the table.
(5) Estimation of Catechin in A. catechu powder by HPTLC
1 2 3 4 5 T 1 2 3 4 5 T
A B
A: In U.V. 254 nm, B: After spraying with vanillin sulphuric acid reagent
Figure: 4.10 HPTLC profile of A. catechu powder
Figure: 4.11 HPTLC chromatogram of standard catechin
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 76
Figure: 4.12 HPTLC chromatogram of standard A. catechu extract
Table: 4.4 Observation table for calibration curve of std. catechin
(Using HPTLC method)
Standard solution
Test
solution
Spot no. 1 2 3 4 5
µl of spot 5 6 7 8 9 6
Concentration
(mcg/µl)
0.5 0.6 0.7 0.8 0.9 -
Peak area 3143.8 3524.1 3822.8 4299.8 4788.5 4230.1
% catechin - - - - - 12.8 %
y = 4065.x + 1070.R² = 0.991
0
1000
2000
3000
4000
5000
6000
0 0.4 0.8 1.2
AU
C
Concentratuion (mcg/µl )
Figure: 4.13 Calibration curve of catechin by HPTLC method
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 77
The concentration of catechin in A. catechu extract was calculated using regression
equation obtained from the standard curve which was found to be 12.5 %.
(6) Estimation of total tannins in A. catechu powder
The total tannins content estimated in A. catechu was 25.36% and it reveals that the
A. catechu powder sample contain total tannins within pharmacopoeial limit which is
25-30 % .
4.2.3 Evaluation of Clove oil
(1) Description
The clove oil was transparent with pale yellow color, aromatic odour and pungent
taste.
(2) Determination of Refractive Index
The Refractive index of clove oil was measured as per IP procedure and it was within
the pharmacopoeial limit as mentioned in Table 4.5.
(3) Determination of Specific Gravity
The Specific Gravity of clove oil was measured as per IP procedure and it was within
the pharmacopoeial limit as mentioned in Table 4.5.
(2) Identification by TLC
In the TLC of clove oil, pink spot at Rf = 0.52 and greenish yellow spot at Rf = 0.52
was observed in U.V. 254 nm and after spraying with Anisaldehyde sulphuric acid
reagent respectively in both extract and std. solution.
Table: 4.5 Results of performed physical parameters of clove oil
Sr. No. Physical parameter Observation Reference value
1. Refractive index 1.534 1.527 - 1.535
2. Specific gravity 1.039 1.38 -1.060
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 78
A B A B
(1) (2)
A: Test solution of clove oil, B: std. solution of eugenol ,
(1): In U.V. 254 nm , (2): After spraying with Anisaldehyde sulphuric acid
Figure: 4.14 TLC profile of clove oil
(3) Estimation of Eugenol in clove oil by HPTLC
1 2 3 4 5 T 1 2 3 4 5 T
A B
A: In U.V. 254 nm, B: After spraying with Anisaldehyde sulphuric acid reagent
Figure: 4.15 HPTLC profile of clove oil
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 79
Figure: 4.16 HPTLC chromatogram of standard Eugenol
Figure: 4.17 HPTLC chromatogram of clove oil
Table: 4.6 Observation table for calibration curve of std. eugenol
(Using HPTLC method)
Standard solution Test
solution
Spot no. 1 2 3 4 5
µl of spot 2.5 5 7.5 10 12.5 6
Concentration
(mcg/µl)
0.2 0.4 0.6 0.8 1.0 -
Peak area 13350.9 15673 17123 18435 19200 13355.6
% Eugenol - - - - - 58.34 %
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 80
y = 7409.1x + 12335R2 = 0.9792
0
5000
10000
15000
20000
25000
0 0.2 0.4 0.6 0.8 1 1.2
AU
C
Concentration (mcg/µl )
Figure: 4.18 Calibration curve of Eugenol by HPTLC method
4.2.4 Standardization of extracts by HPTLC
The extracts of G. glabra and A. catechu was showed 9.71% of 18-β-glycyrhhetinic
acid and 41.14 % of catechin , respectively when standardized by HPTLC method.
4.3 Conclusion
Intended raw materials are subjected here for various quantitative and qualitative
parameters like morphology, microscopy, TLC, ash and extractive value and HPTLC.
The observations were showing satisfactory outcomes for various standardization
parameters.
Chapter: 4 Evaluation of Raw Materials
M. Pharm. (Pharmacognosy) 81
References:
1. Khandelwal K.R., Preliminary phytochemical screening, Practical
Pharmacognosy, Nirali Prakashan, Page no. 125, 149-153.
2. Wagner H. and Bladt S., Plant Drug Analysis, A Thin Layer Chromatography
Atlas ,second edition, page no.170,326
3. Anonymous, “Quality standards of Indian medicinal plants”, Vol -III, Published
by Indian council for medicinal research, 2006, 73.
4. Anonymous, “Indian Herbal Pharmacopoeia”, Indian drug Manufacture’s
Association, Mumbai, 2002, page no.1,89,146.
5. Anonymous, “Quality control methods for medicinal plant materials”, WHO,
Geneva, 2002.
6. Anonymous, “ Indian pharmacopoeia”, , Government of India, Ministry of Health
and family welfare, published by the controller of the publications, Delhi, vol. II,
1996A-96, A-99.
Chapter: 5 Formulation and Evaluation of Gel
5.1 Experimental
5.1.1 Materials
Carbopol- 934P
Distilled water
Triethanolamine
Propylene glycol
Sodium dihydrogen phosphate
Sodium hydroxide
Aqueous extracts of Glycyrrhiza glabra and Acacia actechu
Clove oil
5.1.2 Instruments
Linomat V HPTLC spotter and scanner
Modified Diffusion cell
pH meter
Brookfield viscometer
5.1.3 Preparation of Gel Formulations
The Carbopol-934P gels of Glycyrrhiza glabra, Acacia actechu and clove oil was
prepared by cold method described by schmolka1.The weighed amount of carbopol-
934P (0.8%, 1%, 1.2%, 1.4 %) was placed in a beaker and sufficient amount of water
containing weighed amount of Glycyrrhiza glabra, Acacia catechu extract was added
to obtain a homogenous viscous mixture and kept at room temperature for 24 hours.
After formation of carbopol gel, weighed amount of clove oil was mixed in it with
continuous agitation and obtained gel, stored at ambient temperature prior to its use.
Table: 5.1 Preparation of Gel Formulations
M. Pharm. (Pharmacognosy) 82
Chapter: 5 Formulation and Evaluation of Gel
Ingredients A1 A2 A3 A4 A5
G .glabra extract 1.5 % 1.5 % 1.5 % 1.5 % 1.5 %A.catechu extract 1.5 % 1.5 % 1.5 % 1.5 % 1.5 %Clove oil 1.5 % 1.5 % 1.5 % 1.5 % 1.5 %Carbopol -934P 0.6 % 0.8 % 1 % 1.2 % 1.4 %Propylene glycol 10 % 10 % 10 % 10 % 10 %Tirethanolamine 0.5 % 0.5 % 0.5 % 0.5 % 0.5 %Water (q. s.) 100 ml 100 ml 100 ml 100 ml 100 ml
5.1.4 Preparation of standard calibration curves
(1) Preparation of standard calibration curve of 18-β-glycyrhhetinic acid
1 mg of 18-β-glycyrhhetinic acid was dissolved in 25 ml pH Phosphate buffer (pH
6.8) and extracted with 25 (10+10+5) ml of ether. The ether extract was evaporated
up to 10 ml and filtered. The volume of filtrate was adjusted up to 10 ml with ether.
Standard solution of 2, 3, 4, 5, 6, 7 and 8 µl was applied on a Aluminium-backed
silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was developed up to 85 mm
in Toluene: Ethyl acetate : Methanol: Glacial Acetic Acid (2.7:6.0:1.0:0.3) as mobile
phase at 25º C and the chromatogram was recorded by scanning at 254 nm on a
CAMAG TLC scanner.
(2) Preparation of standard calibration curve of Catechin
1 mg of Catechin was dissolved in 25 ml Phosphate buffer (pH 6.8 ) in and extracted
with 25 (10+10+5) ml of ether. The ether extract was evaporated up to 10 ml and
filtered. The volume of filtrate was adjusted up to 10 ml with ether. Standard solution
of 5, 10, 15, 20, 25, 30 and 35 µl was applied on a Aluminium-backed silica gel 60F254
plate (E. Merck) (10 x 10 cm). The plate was developed up to 85 mm in Toluene:
Ethyl acetate: Methanol: Glacial Acetic Acid (2.7:6.0:1.0:0.3) as mobile phase at 25º
C and the chromatogram was recorded by scanning at 254 nm on a CAMAG TLC
scanner.
5.1.5 Evaluation of gel formulations 2-9
a) pH
M. Pharm. (Pharmacognosy) 83
Chapter: 5 Formulation and Evaluation of Gel
1 gm of gel was accurately weighed and dispersed in 10 ml of distilled water. The pH
of these dispersions was measured using pH meter (Systronics digital- DI- 707).
b) Viscosity and Rheological studies
Viscosities of gels were determined using Brookfield Viscometer. Gels were tested
for their Rheological characteristics at 25oC using Brookfield Viscometer (DV-III
programmable Rheometer). The Measurement was made over the whole range of
speed setting from 10 rpm to 100 rpm with 30 seconds between two successive speeds
and then in descending order.
c) Spreadibility
For the determination of spreadibility excess of sample was applied in between two
glass slides and was compressed to uniform thickness by placing 1000 gm weight for
5 minutes. Weight (50 gm) was added to the pan. The time required to separate the
two slides i.e. the time in which the upper glass slide moves over the lower plate was
taken as measure of spreadibility (S).
S= m × l/t
Where,
m = weight tide to upper slide
l = length moved on the glass slide
t = time taken
d ) Drug Content
For the determination of drug content , 1 gm of gel from each formulation was taken
and dissolved in 25 ml Phosphate buffer (pH 6.8) and extracted with 25 (10+10+5) ml
of ether. The ether extract was evaporated up to 10 ml and filtered. The volume of
filtrate was adjusted up to 10 ml with ether. 5 µl of this solution was applied on a
Aluminium-backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was
developed up to 85 mm in Toluene: Ethyl acetate: Methanol: Glacial Acetic Acid
(2.7:6.0:1.0:0.3) as mobile phase at 25º C and the chromatogram was recorded by
scanning at 254 nm on a CAMAG TLC scanner. The concentration of 18-β-
glycyrhhetinic acid and catechin in the gel was determined using the calibration curve
of standard 18-β-glycyrhhetinic acid and catechin respectively.
M. Pharm. (Pharmacognosy) 84
Chapter: 5 Formulation and Evaluation of Gel
e) In vitro Release study
In-vitro diffusion studies were carried out in fabricated diffusion tube of surface area
1.5 cm2 through cellophane membrane. The cellophane membrane was hydrated for
24 hours with 0.1N NaOH and 1 hour with distilled water, 1gm of gel was applied on
it and fixed to the one end of the tube which act as a donor compartment. The
assembly was placed in the beaker contained 50 ml of Phosphate buffer (pH 6.8). The
teflon coated magnetic bead was placed in the beaker and rotated at 100 rpm using
magnetic stirrer and the temperature was maintained at 37 1 0 C.
Samples of 3 ml were withdrawn at regular intervals of every one hrs up to 8
hrs and replace the volume with same buffer and maintained sink condition through
the studies. The samples were extracted with 5 ml of ether and filtered. The volume of
filtrate was adjusted up to 5 ml with ether. 25 µl of this solution was applied on a
Aluminium-backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was
developed up to 85 mm in Toluene: Ethyl acetate: Methanol: Glacial Acetic Acid
(2.7:6.0:1.0:0.3) as mobile phase at 25º C and the chromatogram was recorded by
scanning at 254 nm on a CAMAG TLC scanner. The concentration of 18-β-
glycyrhhetinic acid and catechin in the gel was determined using the calibration curve
of standard 18-β-glycyrhhetinic acid and catechin respectively.
5.2 Results and Discussion
5.2.1 Preparation of gel formulations
M. Pharm. (Pharmacognosy) 85
Chapter: 5 Formulation and Evaluation of Gel
Gels prepared with carbopol were found to be translucent and homogeneous with
characteristics colour of extracts of Glycyrrhiza glabra and Acacia catechu.
Figure 5.1: Prepared Gel Formulation
5.2.2 Preparation of standard calibration curves
The calibration curves of 18-β-glycyrhhetinic acid and catechin were prepared as per
the procedure given in section 5.1.4 and are mention below.
y = 7720.2x - 1163
R2 = 0.9934
0
1000
2000
3000
4000
5000
6000
0 0.2 0.4 0.6 0.8 1
Concentration (mcg/µl)
AU
C
Figure: 5.2 Calibration curve of 18-β-G.A. by HPTLC method
M. Pharm. (Pharmacognosy) 86
Chapter: 5 Formulation and Evaluation of Gel
y = 4697.2x + 953.1
R2 = 0.9932
0
4000
8000
12000
16000
20000
0 0.5 1 1.5 2 2.5 3 3.5 4
Concentration (mcg/µl)
AU
C
Figure: 5.3 Calibration curve of Catechin by HPTLC method
5.2.3 Evaluation of Gel Formulations
Evaluation parameters of prepared gels have been carried out as per the procedure
given in section 5.1.5 and show in Table 5.2 and 5.3.
M. Pharm. (Pharmacognosy) 87
Table: 5.2 Evaluation Parameters of Gels
Formulation
codepH
Viscosity
(cps)
Spreadibility
(gm*cm/s)Drug content (%)
18-β-G.A. CatechinA1 6.65 587.33 × 103 3.21 84.65 71A2 6.47 747.3 × 103 1.19 80.0 73A3 6.35 124 × 104 0.93 79.5 70A4 6..27 136.33 × 104 0.81 83.2 72.65A5 6.10 145.66 × 104 0.74 81.7 68
Chapter: 5 Formulation and Evaluation of Gel
1 2 3 4 Catechin Eugenol 18-β.G.A
1, 2, 3, 4 :spot of gel
Figure: 5.4 HPTLC profile of Gel
Figure: 5.5 HPTLC chromatogram of gel
M. Pharm. (Pharmacognosy) 88
Chapter: 5 Formulation and Evaluation of Gel
Figure: 5.6 HPTLC chromatogram of standard 18-β-G.A.
Figure: 5.7 HPTLC chromatogram of standard catechin
M. Pharm. (Pharmacognosy) 89
Chapter: 5 Formulation and Evaluation of Gel
Figure: 5.8 HPTLC chromatogram of standard Eugenol
In the prepared gel formulations, drug content of 18-β-glycyrrhitinic acid was within
79.5-84.65 and of catechin was 68-73 % (Table 5.2) in the formulations. Viscosity,
Spreadibility and pH were measured for different formulation are show in Table 5.2.
The Formulation A4 had viscosity 136.33 × 104 which was easy to spread.
In vitro release profile from gel formulations
The data obtained from in vitro release (Q1 and Q6) are show in Table 5.3.
Table: 5.3 In vitro Release profile of gel formulationsFormulation
code18-β-G.A. Catechin
Q1 Q6 Q1 Q6
A1 6.4 71.2 4.1 71A2 8.6 77.4 4.6 73A3 9.2 80.1 6.3 70A4 10.1 85.6 7.2 72.65A5 10.5 86.8 8.1 68
Q1 : In vitro Release of Drug in 1 hr
Q6 : In vitro Release of Drug in 6 hr
M. Pharm. (Pharmacognosy) 90
Chapter: 5 Formulation and Evaluation of Gel
0
20
40
60
80
100
0 2 4 6 8 10
% CPR of 18
-18-β.
G.A.
Time (hr)
A1 A2 A3 A4 A5
Figure: 5.9 In vitro release profile of 18-β-G.A. in gel
in phosphate buffer (pH 6.8 ) at 254 nm
0
20
40
60
80
100
0 2 4 6 8 10
% CPR for Catechin
Time (hr)
A1 A2 A3 A4 A5
Figure: 5.10 In vitro release profile of catechin in gel
in phosphate buffer (pH 6.8)at 254 nm
5.3 Conclusion:
From this study it was revealed that as the concentration of the polymer increase , the
various parameters such as viscosity, spreadibility, pH, drug content and in vitro
release was also changed. The viscosity of the formulation was increased and
spreadibility decrease with increasing the concentration of polymer in the
formulations. The formulations were shown maximum in vitro release at 6 hr and
after that the release was constant. According to data shown in Table 5.2 for pH,
M. Pharm. (Pharmacognosy) 91
Chapter: 5 Formulation and Evaluation of Gel
viscosity, spreadibility, Drug content and In vitro Release study, it was concluded that
the formulation A4 was better than other formulations in respect to in vitro release
profile and viscosity.
References:
1. Schmolka, I.S, J. Biomed. Maater. Res., 1972, 6, 571.
2. Saleem, M. A., Sanaullah, S. and Purohit, M. G., Antimicrobial and wound
healing activity of prepared gatifloxacin topical gels, Indian Drugs, 2005, 43(4),
292-295.
3. Kikwai, L., Babu, Jayachandra, Prado, R., Kolot, A., Cheryl A., Armstong, Ansel,
John, C. and Singh Mandeep, Invitro and invivo evaluation of topical formulations
of spantide II, AAPS PharmSciTech, 2005, 6(4), EF 65-72.
4. Nayak, S. H., Nakhat, P. D. and Yeole, P. G., Development and evaluation of
cosmeceutical hair styling gels of ketoconazole, Indian J. Pharm. Sci., 2005, 313-
316.
5. Kavitha, K., Sivaramakrishnan, M. and Nalini, C. N., Formulation and evaluation
of topical drug delivery system of fluconazole, Indian Drugs, 2003, 40 (12), 720-
723.
6. Djordjevic, J., Michniak, B. and Uhrich, Kathryn E., Amphiphilic star like
macromolecules as novel carriers for topical delivery of non steroidal anti-
inflammatory drugs, AAPS PharmSciTech, 2003, 5(4), 1-12.
7. Ota, Yusuke, Hamada, A., Nakano. M. and Saito, H., Evaluation of percutaneous
absorption of midazolam by terpenes, Drug Metab. Pharmacokin, 2003, 18(4),
261-266.
M. Pharm. (Pharmacognosy) 92
Chapter: 5 Formulation and Evaluation of Gel
8. Uma devi, S., Ganesan, M. and Mohanta, G. P., Design and evaluation of
tetracycline hydrochloride gels, Indian Drugs, 2002, 39(10), 552-554.
9. Raghuramana.S. et.al., In Design and evaluation of propranolol hydrochloride
buccal films. Indian J. Pharm. Sci., 2002, 64 (1): 32-36.
M. Pharm. (Pharmacognosy) 93
Chapter: 6 Formulation and Evaluation of Patch
6.1. Experimental:
6.1.1. Materials:
HPMC K15M
Distilled water
Methanol
Glycerine
Sodium dihydrogen phosphate
Sodium hydroxide
Aqueous extracts of Glycyrrhiza glabra and Acacia actechu
Clove oil
6.1.2. Instruments:
Linomat V HPTLC spotter and scanner
Modified Diffusion cell
pH meter
USP disintegration apparatus
Digital vernier caliper
6.1.3. Preparation of patch formulations 1
The patches were prepared by solvent evaporation technique in Petridish. The
weighed amount of Glycyrrhiza glabra and Acacia catechu extracts were dissolved in
sufficient amount of distilled water. The weighed amount of clove oil was mixed with
sufficient amount of methanol and mixed with above mixture. The weighed amount of
HPMC K15M (0.5%, 1 %, 1.5%) was added in the mixture under stirring condition
with a magnetic stirrer. Glycyrine was added as plasticizer. The dispersion was
poured on a petridish and dried at room temperature for 24 hrs. the prepared patches
were packed in aluminum foil.
Table 6.1 : Preparation of patch formulations
M. Pharm. (Pharmacognosy) 93
Chapter: 6 Formulation and Evaluation of Patch
Ingredients B1 B2 B3
G.glabra extract 2 mg 2 mg 2 mgA.catechu extract 2 mg 2 mg 2 mgClove oil 6 µl 6 µl 6 µlHPMC K15M 0.5 % 1 % 1.5 %Glycerine 0.2 ml 0.2 ml 0.2 mlWater 5 ml 5 ml 5 mlMethanol 5 ml 5 ml 5 ml
6.1.4. Preparation of standard calibration curve:
(1) Preparation of standard calibration curve of 18-β-glycyrhhetinic acid
1 mg of 18-β-glycyrhhetinic acid was dissolved in 25 ml Phosphate buffer (pH 6.8) in
and extracted with 25 (10+10+5) ml of ether. The ether extract was evaporated up to
10 ml and filtered. The volume of filtrate was adjusted up to 10 ml with ether.
Standard solution of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7 and 0.8 µl was applied on a
Aluminium-backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was
developed up to 85 mm in Toluene: Ethyl acetate : Methanol: Glacial Acetic Acid
(2.7:6.0:1.0:0.3) as mobile phase at 25º C and the chomatogram was recorded by
scanning at 254 nm on a CAMAG TLC scanner.
(2) Preparation of standard calibration curve of catechin
1 mg of catechin was dissolved in 25 ml Phosphate buffer (pH 6.8) in 100 ml beaker
and extracted with 25 (10+10+5) ml of ether. The ether extract was evaporated up to
10 ml and filtered. The volume of filtrate was adjusted up to 10 ml with ether.
Standard solution of 0.5, 1, 1.5, 2, 2.5 ,3 and 3.5 µl was applied on a Aluminium-
backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was developed up to
85 mm in Toluene: Ethyl acetate : Methanol: Glacial Acetic Acid (2.7:6.0:1.0:0.3) as
mobile phase at 25º C and the chomatogram was recorded by scanning at 254 nm on a
CAMAG TLC scanner.
6.1.5 Evaluation of gel formulations 2,3,4
a) Patch Thickness
M. Pharm. (Pharmacognosy) 94
Chapter: 6 Formulation and Evaluation of Patch
Thickness of patches of was measured using digital vernier caliper at three different
places and the mean value was calculated.
b) Determination of Surface pH
For determination of surface pH, the patches (14 mm) of each formulation were kept
in contact with 1 ml of distilled water for 2 hr, in fabricated glass tubes. Excess of
water from the tubes was drained and the pH was noted by bringing the electrodes
near the surface of the formulation and allowing it to equilibrate for 1 min.
c) Folding Endurance:
The patches of each formulation of size (2× 2 cm) were cut by using sharp blade.
Folding endurance was determined by repeatedly folding a small strip of patch at the
same place till it broke. The number of times the patch could be folded at the same
place without breaking gave the value of folding endurance.
d) In vitro Residence time
The in vitro residence time was determined using USP disintegration apparatus. The
disintegration medium was 800 ml of Phosphate buffer (pH 6.8) maintained at
37±2°C. The Segment of rat intestinal mucosa, each of 3 cm length, was glued to the
surface of glass slab, which was then vertically attached to the apparatus. Three
mucoadhesive patches of each formulation were hydrated on one surface using pH 6.8
PB and the hydrated surface was brought into contact with the mucosal membrane.
The glass slab was vertically fixed to the apparatus and allowed to move up and
down. The patch was completely immersed in the buffer solution at the lowest point
and was out at the highest point. The time required for complete erosion or
detachment of the patch from the mucosal surface was recorded.
e) Drug Content
For the determination of drug content , 1.5 × 1.5 cm of patch from each formulation
was taken and dissolved in 10 ml Phosphate buffer (pH 6.8) and extracted with 10
(5+5) ml of ether. The ether extract was evaporated up to 5 ml and filtered. The
volume of filtrate was adjusted up to 5 ml with ether. 20 µl of this solution was
applied on a Aluminium-backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The
M. Pharm. (Pharmacognosy) 95
Chapter: 6 Formulation and Evaluation of Patch
plate was developed up to 85 mm in Toluene: Ethyl acetate: Methanol: Glacial Acetic
Acid (2.7:6.0:1.0:0.3) as mobile phase at 25º C and the chromatogram was recorded
by scanning at 254 nm on a CAMAG TLC scanner. The concentration of 18-β-
glycyrhhetinic acid and catechin in the gel was determined using the calibration curve
of standard 18-β-glycyrhhetinic acid and catechin respectively.
f) In vitro Release study
In-vitro diffusion studies were carried out in fabricated diffusion tube of surface area
1.5 cm2 through cellophane membrane. The cellophane membrane was hydrated for
24 hours with 0.1N NaOH and 1 hour with distilled water, 0.5 × 0.5 cm of patch from
each formulation was applied on it and fixed to the one end of the tube which act as a
donor compartment. The assembly was placed in the beaker contained 50 ml of
Phosphate buffer (pH 6.8). The teflon coated magnetic bead was placed in the beaker
and rotated at 100 rpm using magnetic stirrer and the temperature was maintained at
37 1 0 C.
Samples of 3 ml were withdrawn at regular intervals of every one hrs up to 8
hrs and replace the volume with same buffer and maintained sink condition through
the studies. The samples were extracted with 5 ml of ether and filtered. The volume of
filtrate was adjusted up to 5 ml with ether. 35 µl of this solution was applied on a
Aluminium-backed silica gel 60F254 plate (E. Merck) (10 x 10 cm). The plate was
developed up to 85 mm in Toluene: Ethyl acetate: Methanol: Glacial Acetic Acid
(2.7:6.0:1.0:0.3) as mobile phase at 25º C and the chromatogram was recorded by
scanning at 254 nm on a CAMAG TLC scanner. The concentration of 18-β-
glycyrhhetinic acid and catechin in the gel was determined using the calibration curve
of standard 18-β-glycyrhhetinic acid and catechin respectively.
6.2 Results and Discussion
6.2.1 Preparation of patch formulations
M. Pharm. (Pharmacognosy) 96
Chapter: 6 Formulation and Evaluation of Patch
Patches prepared with HPMC were found to be translucent, smooth and with
characteristics color of extracts of Glycyrrhiza glabra and Acacia catechu.
Figure 6.1: Prepared Patch Formulation
6.2.2 Preparation of standard calibration curves
The calibration curves of 18-β-glycyrhhetinic acid and catechin were prepared as per
the procedure given in section 6.1.4 and are mention below.
y = 7720.2x - 116.3
R2 = 0.9934
0
100
200
300
400
500
600
0 0.02 0.04 0.06 0.08 0.1
Concentration (mcg/ µl)
AU
C
Figure: 6.2 Calibration curve of 18-β-G.A.by HPTLC method
M. Pharm. (Pharmacognosy) 97
Chapter: 6 Formulation and Evaluation of Patch
y = 4697.2x + 95.309
R2 = 0.9932
0
400
800
1200
1600
2000
0 0.1 0.2 0.3 0.4
Concentration (mcg/µl)
AU
C
Figure: 6.3 Calibration curve of Catechin by HPTLC method
6.2.3 Evaluation of Patch Formulations
Evaluation parameters of prepared patches have been carried out as per the procedure
given in section 6.1.5 and show in Table 6.2 and 6.3.
1 2 3 4 Catechin Eugenol 18-β.G.A
M. Pharm. (Pharmacognosy) 98
Table: 6.2 Evaluation Parameters of PatchesFormula
-tion
code
Patch
Thickness
(mm)
Surface
pH
In vitro
residence
time (hr)
Folding
Endurance
Drug content
(mg/cm2)
18-β-G.A. catechinB1 0.10 ± 0.015 6.15 2.50 150 4.08×10-3 4.08×10-3
B2 0.14 ± 0.021 6.37 2.75 159 6.12×10-3 6.12×10-3
B3 0.16 ± 0.015 6.42 3.15 168 7.64×10-3 7.64×10-3
Chapter: 6 Formulation and Evaluation of Patch
1, 2, 3, 4:spot of patch
Figure: 6.4 HPTLC profile of Patch
Figure: 6.5 HPTLC chromatogram of Patch
Figure: 6.6 HPTLC chromatogram of standard 18-β-G.A.
M. Pharm. (Pharmacognosy) 99
Chapter: 6 Formulation and Evaluation of Patch
Figure: 6.7 HPTLC chromatogram of standard Catechin
Figure: 6.8 HPTLC chromatogram of standard Eugenol
In the prepared patch formulations, Drug content of 18-β-glycyrrhitinic acid was
within 4 × 10-3 -7.7 × 10-3/ cm2 and of catechin was 0.016-0.033/ cm2 (Table 6.2).
Patch Thickness, Surface pH , In vitro residence time and Drug content were
measured for different formulation was shown in Table 6.2.
In vitro release profile from patch formulations
The data obtained from in vitro release study are show in Table 6.3.
Table: 6.3 In vitro Release profile of patch formulationsFormulation
code18-β-G.A. Catechin
Q0.5 Q3 Q0.5 Q3
B1 8.9 75.8 7.6 71.8
M. Pharm. (Pharmacognosy) 100
Chapter: 6 Formulation and Evaluation of Patch
B2 13.5 79.3 8.6 75.1B3 15.3 83.2 12.22 80.0
Q0.5: In Vitro Release of drug in 0.5 hr
Q3: In Vitro Release of drug in 3 hr
0
20
40
60
80
100
0 1 2 3 4 5 6
% CPR of18
-β.G.A
Time (hr)
B1 B2 B3
Figure: 6.9 In vitro release profile of 18-β-G.A. in patch
in Phosphate buffer (pH 6.8) at 254 nm
0
20
40
60
80
100
0 1 2 3 4 5 6
% CPR of Catechin
Time (hr)
B1 B2 B3
Figure: 6.10 In vitro release profile of catechin in patch
in Phosphate buffer (pH 6.8) at 254 nm
6.1 Conclusion:
M. Pharm. (Pharmacognosy) 101
Chapter: 6 Formulation and Evaluation of Patch
From this study it was revealed that the as the concentration of the polymer , the
various parameters such as surface pH, in vitro residence time, drug content and in
vitro diffusion was also changed. The In vitro residence time of the formulation was
increased with increasing the concentration of polymer in the formulations. The
formulations were shown maximum in vitro diffusion at 3 hr and after that the
diffusion was constant. The selection of best formulation depends on the In vitro
residence time and In vitro Release study. The in vitro Release was higher in the
formulation B3 as compared to other formulations. The In vitro residence time of
formulation B3 was also higher as compared to other formulations and it was retain
on the mucosal surface up to the maximum diffusion of the drug . So, formulation B3
was selected as the best formulation.
References:
1. Ting Li et. Al., Optimized preparation and evaluation of Indomethacin
Transdermal Patch, Asian Journal of Pharmaceutical Sciences, 2007, 2 (6): 249-
259.
2. Semalty M., Semalty A., Kumar G., Formulation and Characterization of
Mucoadhesive Buccal Films of Glipizide, Indian Journal of Pharmaceutical
Sciences, Jan-Feb 2008, 43-48.
3. Sahni J., Raj S., Ahmad F.J. Khar R.K., Design and In Vitro Characterization of
Buccoadhesive Drug Delivery System of Insulin, Jan-Feb 2008, 61-65.
4. Raghuramana.S. et.al., In Design and evaluation of propranolol hydrochloride
buccal films. Indian J. Pharm. Sci., 2002, 64 (1): 32-36.
M. Pharm. (Pharmacognosy) 102
Chapter: 6 Formulation and Evaluation of Patch
M. Pharm. (Pharmacognosy) 103
Chapter: 7 Antimicrobial Activity of selected Formulations
7.1 Introduction 1
Pharmaceutical compositions are provided which comprise effective amounts of
antimicrobials, anti-inflammatories, and antihistamines, to provide an ulcer
medication which prevents secondary infections and promotes healing while
providing immediate relief from pain. Antimicrobial compounds can be used to
prevent the ulcer from spreading and relief.
In some cases there are infectious agents that are both bacterial and viral in nature that
are considered as causes of mouth ulcers. The various chemical compounds that are
found within the infectious agents are perhaps one of the reasons for mouth ulcers
forming.
7.2 Material and method 2-6
Test organisms and Inoculums:
Gram positive:- Bacillus subtilis, Staphylococcus aureus
Gram negative :- Escherichia coli, Pseudomonas aeruginosa
Standard:- Antimicrobial Nitrofurazone gel (500mg).
Media: - Dehydrated nutrient agar media was used and was prepared in distilled
water. The composition of the media was as given below.
Composition of nutrient agar medium
1) Agar 15.0%
2) Peptic digest of animal tissue 5.0%
3) Sodium chloride 5.0%
4) Beef extract 1.5%
5) Yeast extract 1.5%
6) pH 7.4 ± 0.2 at 25oC
7) distilled water 1000 ml
The medium was autoclaved at 15 lbs per square inch pressure at 121oC
Preparation of media: - Dehydrated nutrient agar media (28gm) was
accurately weighed and suspended in 1000 ml of distilled water in a conical
flask. It was heated on a water bath to dissolve the medium completely.
M. Pharm. (Pharmacognosy) 104
Chapter: 7 Antimicrobial Activity of selected Formulations
Sterilization of media: - The conical flask containing the nutrient agar
medium was plugged with the help of non-absorbent cotton bung. The mouth
of the conical
flask and the cotton bung were properly covered with aluminum foil. The medium
was then sterilization by autoclaving at 15 lbs per square inch pressure for 20 minutes.
Method: - Cup and Plate method. The sterile nutrient agar medium at a
temperature between 40o to 50o C was immediately poured into the sterile Petri
plates to give a depth of 3 to 4 mm, by placing the plates on a level surface.
The plates were then allowed to solidify. Each plate was then inoculated with
0.1ml of the solution of test organisms prepared in water for injection. The
wells in each plate were bored in the centre that was filled with 500 mg plain
gel, 300,500 and 700 mg of selected gel (A4) and 0.5 cm2 plain patch , 0.5 cm2
and 1.0 cm2 of selected patch (B3). The plates were then incubated at 37o for
24h. After incubation, zonal inhibition (inhibition around each well) was
measured and this value was taken as an indicator for the antimicrobial
activity.
7.3 Result and Discussion
7.3.1 Result of Antimicrobial activity of selected Gel Formulations
(a) (b)
M. Pharm. (Pharmacognosy) 105
Chapter: 7 Antimicrobial Activity of selected Formulations
(c) (d)
Figure: 7.1 Antimicrobial Activity of gel formulation; (a) Bacillus subtilis ,
(b) Staphylococcus aureus, (c) Pseudomonas aeruginosa , (d) Escherichia coli
The zone of inhibition of gel formulations was mentioned below in Table 7.1
7.3.2 Result of Antimicrobial activity of selected Patch Formulations
(a) (b)
(c) (d)
M. Pharm. (Pharmacognosy) 106
Table: 7.1 Zone of Inhibition (cm) of gel formulationNitrofurazone
gel
Plain
gel0.3 mg gel 0.5 mg gel 0.7 mg gel
Bacillus subtilis 2.2 0 1.8 1.7 2.5Escherichia coli 2.0 0 2 2.4 2.7Pseudomonas
aeruginosa2.7 0 1.9 2.2 2.6
Staphylococcus
aureus2.0 0 2.5 2.7 2.9
Chapter: 7 Antimicrobial Activity of selected Formulations
Figure: 7.2 Antimicrobial Activity of patch formulation; (a) Bacillus subtilis ,
(b) Staphylococcus aureus, (c) Pseudomonas aeruginosa , (d) Escherichia coli
The zone of inhibition of patches formulations was mentioned below in Table 7.2
7.4 Conclusion :
The selected formulation of gel and patch was evaluated for antimicrobial activity
against various gram positive and gram negitive bacteria. the plain gel and patch
without the herbal extracts did ot shown any zone of inhibition .The formulations
were shown the significant antimicrobial activity against various bacteria and which
would be benificial in the Mouth ulcer.
References:
1. Campbell, Phillip, Lesion and ulcer medication ,United States Patent 6352711
2. Anonymous, “British pharmacopoeia”, 2002, Pharmaceutical press, London, 796.
3. Anonymous, “Indian pharmacopoeia”, 1996, The Controller of Publications,
Delhi, Vol. II, A-105.
4. Vivek K. Gupta, Atiya Fatima, Uzma Faridi, Arvind S. Negi, Karuna Shanker,
J.K. Kumar, , Journal of Ethnopharmacology, 5 March 2008,116( 2) , 377-380.
5. G.A. Ayoola, African Journal of Microbiology Research. July 2008, 2 , 162-166
6. Li ZhongXing, Wang XiouHua, Yue YunSheng, Zhao BaoZhen, Chen JingBo, Li
JiHong, Chinese Journal of Information on Traditional Chinese Medicine.
M. Pharm. (Pharmacognosy) 107
Table:7.2 Zone of Inhibition (mm) of patch formulationPlain patch 0.5cm2 patch 1.0 cm2 patch
Bacillus subtilis 0 1 1.6Escherichia coli 0 1.2 1.2Pseudomonas aeruginosa 0 0.8 0.9Staphylococcus aureus 0 0.9 1
Chapter: 7 Antimicrobial Activity of selected Formulations
M. Pharm. (Pharmacognosy) 108