REVIEW OF LITERATURE - Shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/73024/9/08. chapter 2.pdf · linked to skin aging and is co-related with an increase in activity of certain

REVIEW OF LITERATURE

School of Science, SVKM’s NMIMS University Page 21

CHAPTER 2

REVIEW OF

LITERATURE



2. REVIEW OF LITERATURE

2.1 IN VITRO ASSAYS FOR EVALUATION OF ANTI-AGING ACTIVITY

2.2 IN VITRO ASSAYS FOR EVALUATION OF ANTI- ACNE ACTIVITY

2.3 TLC-BIOAUTOGRAPHY METHOD

2.4 REVIEW OF LITERATURE OF PLANTS SELECTED FOR THE STUDY

2.4.1 Ocimum tenuiflorum Linn

2.4.2 Citrus reticulata Blanco

2.4.3 Citrus aurantifolia (Christm) Swingle

2.4.4 Butea monosperma Lam

2.4.5 Vitis vinifera Linn



2.1 In vitro assays for evaluation of anti-aging activity

Skin is a major candidate and target of oxidative stress. The skin aging is an inevitable

process for all living organisms. During this processs, ROS generation is increased

which leads to the degradation of ECM. Degradation of the ECM has directly been

linked to skin aging and is co-related with an increase in activity of certain enzymes

involved in skin aging. These enzymes are mainly elastase and collagenase (Ndlovu et

al. 2013). Topical application of antioxidants with specific inhibition activity of

enzymes for protection of ECM can be a useful approach to prevent the skin damage

from various sources. Furthermore, plant sources have been evaluated for developing

natural antioxidants that may be involved in anti-ageing and anti- wrinkle care

(Oresajo et al. 2010)

Recently, a number of studies focused on the determination of anti-aging activity of

plant extracts through in vitro antioxidant, anti-collagenase and anti-elastase activity.

Thring et al studied the anti-collagenase, anti-elastase and anti-oxidant activities of

extracts from 21 plants and shown that anti-collagenase activities were exhibited by

sixteen plants, of which the highest activity was seen in white tea (~87%) (Thring et

al. 2009). Chompoo et al demonstrated the effect of Alpinia zerumbet components on

antioxidant and skin diseases-related enzymes. Results indicate that the rhizome

aqueous extract exhibited greater collagenase inhibitory activity and proved to be the

source of bioactive compounds against enzymes responsible for causing skin diseases

(Chompoo et al. 2012). In a very recent study, Wahab et al assessed antioxidant

capacity, anti-collagenase and anti-elastase activity of Malaysian unfermented cocoa

bean for cosmetic application (Wahab et al. 2014). A number of studies have been

interested in interactions between elastase and its inhibitors (Edwards & Bernstein

1994) (Bizot-Foulon et al. 1995) (Vasconcelos et al. 2011). Kim at al investigated the

inhibitory effects of 150 medicinal plants on elastase activity. In vitro screening of

Jeju medicinal plants were performed in the search of new cosmeceutical materials. In

this study, extracts obtained from 254 different kinds of Jeju medicinal plants were

screened for inhibitory effects on tyrosinase and elastase, and for free radical

scavenging effects. Results demonstrated that 17 plant extracts were able to inhibit

elastase enzyme and thus can be valuable in the development of anti-aging cosmetics

(Kim et al. 2007).



The list of plants showing anti-aging activity by invitro antioxidant and enzyme

assays is represented in Table 2.1.

Table 2.1: List of anti-aging plants and in vitro methods employed

Sr.

No Name of the Plants

Methods

employed References

1

Clerodendrum glabrum,

Peltophorum africanum,

Psychotria capensis,

Schotia brachypetala

Antioxidant assay:

ABTS radical scavenging

assay

Anti-elastase assay,

Anti-collagenase assay,

Anti-hyaluronidase assay

(Ndlovu et al.

2013)

2 Alpinia zerumbet

Antioxidant assay:

DPPH, ABTS, Superoxide

assay,

Anti-collagenase assay,

Anti-elastase assay,

Anti-hyaluronidase assay,

(Chompoo et al.

2012)

3 Albizzia julibrissin

Durazz.

Antioxidant assays:

DPPH assay

Nitrite scavenging assay,

Elastase inhibition assay,

(Leem 2011)

4 Garcinia indica

Hyaluronidase inhibition

assay,

Elastase inhibition assay

(Sahasrabudhe &

Deodhar 2010)

5 Areca catechu L


assay,

Elastase inhibition assay

(Lee et al. 2001)

6 Tagetes erecta Linn


assay,

Elastase inhibition assay,

Matrix Metalloproteinase

(MMP-1) inhibition assay

(Maity et al.

2011)



2.2 In vitro assays for evaluation of anti-acne activity

Acne vulgaris is a disease of pilosebaceous follicles. It generally begins in puberty,

when androgen levels increase significantly and stimulate excess sebum secretion

(Scholl et al. 1984). It is a multifactorial disease. Especially colonization of P. acnes

in sebaceous gland is one of the causative factors for acne vulgaris. Apart from P.

acnes, S. epidermidis and Staphylococcus aureus (S. aureus) also play major role in

pathophysiology of acne. P. acnes has been described as an inflammatory anaerobic

organism that is implicated in the development of inflammatory acne, while S.

epidermidis and S. aureus are aerobic organisms that are usually involved with

superficial infections of the sebaceous unit (Burkhart et al. 1999). Hassanzadeh et al

showed that the most frequent bacteria isolated from acne patients was S. aureus

(Hassanzadeh et al. 2008). Several reports showed that, the secondary metabolite

products of medicinal plants have been effectively used as a source of antimicrobial

agents against acne inducing bacteria (Lertsatitthanakorn et al. 2006) (Park et al.

2004) (Saising & Voravuthikunchai 2012)

Antimicrobial activity of plant extracts and their mixtures can be determined by

various diffusion and dilution methods. Agar well diffusion method is commonly used

for preliminary screening of antimicrobial activity followed by the determination of

Minimum inhibitory concentration (MIC) and Minimum bactericidal concentration

(MBC) of test extracts using broth dilution or agar dilution methods (Klančnik et al.

2010). Chomnawang et al studied the antimicrobial effects of Thai medicinal plants

against acne-inducing bacteria by disc diffusion and broth dilution methods

(Chomnawang et al. 2005). Ali-Shtayeh et al investigated the antimicrobial activities

of 56 Palestinian medicinal plants against etiologic agents of acne vulgaris, mainly P.

acnes and S. aureus using disc diffusion and broth dilution methods (Ali-shtayeh et al.

2013). Time kill efficacy study is a measure of time dependant antibacterial activity.

Usually broth dilution methods are adapted for determination of time kill curves.

Olajuyigbe and Afolayan studied the in vitro antibacterial and time-kill assessment of

crude methanolic stem bark extract of Acacia mearnsii De Wild against Gram

positive and Gram negative bacteria (Olajuyigbe & Afolayan 2012).



P. acnes lipase is an important factor in the pathogenesis of acne because it

breakdowns the sebaceous triglycerides in to free fatty acids which leads to severe

inflammation (Higaki 2003). If plant extract is able to inhibit this lipase enzyme then

it will be beneficial as anti-acne agent. Batubara et al screened the antiacne potency of

Indonesian medicinal plants by antibacterial, lipase inhibition, and antioxidant assays

(Batubara et al. 2009). Patil et al demonstrated the anti-lipase activity of Indian

medicinal plants by colorimetric microasssay and plate assay method (Patil et al.

2012).

The list of plants showing anti-acne activity by antimicrobial methods is represented

in Table 2.2.

Table 2.2: List of anti-acne plants and in vitro methods employed

Sr.

No Name of the Plants Methods

Employed

Test

organisms References

1

Rhodomyrtus tomentosa

(Aiton) Hassk

Disc diffusion,

Broth dilution,

Time-kill curves,

Cytotoxicity

assay

P. acnes

(Saising &

Voravuthikunc

hai 2012)

2

Picrorhiza kurroa,

Vitex negundo,

Terminalia chebula,

Embelia ribes

Agar well

diffusion,

Broth dilution,

Anti-lipase assay

P. acnes

(Patil et al.

2012)

3

Barleria prionitis,

Butea monosperma,

Casuarina equisetifolia,

Dalbergia sympathetic,

Lagenaria siceraria

Agar well

diffusion,

Antioxidant assay

P. acnes

(Thube & Patil

2012)

4

Azadirachta indica,

Vitex negundo,

Annona squamosa,

Cymbopogon citratus

Terminalia chebula

Disc diffusion

assay P. acnes

(Balakrishnan

et al. 2011)



5 Garcinia mangostana

Linn

Broth dilution

assay,

TLC-

Bioautography

method

P. acnes,

S. epidermidis

(Pothitirat et

al. 2010)

6

Mollugo pentaphylla,

Angelica anomala,

Matteuccia orientalis,

Orixa japonica

Disc diffusion

assay,

Broth dilution

assay

P. acnes,

S. epidermidis

(Kim et al.

2008)

7

Hemidesmus indicus,

Eclipta alba,

Coscinium fenestratum,

Curcubito pepo,

Tephrosia purpurea,

Mentha piperita,

Pongamia pinnata,

Symplocos racemosa,

Euphorbia hirta,

Tinospora cordyfolia,

Thespesia populnea,

Jasminum officinale

Disc diffusion

assay,

Broth dilution

assay,

TLC-

Bioautography

method

P. acnes,

S. epidermidis

(Kumar et al.

2007)

8

Excoecaria

cochinchinensis Lour,

Salvia officinalis Lour,

Argyreia nervosa

(Burm.f) Bojer

Agar well

diffusion assay,

Broth dilution

assay

P. acnes,

S. aureus

(Leelapornpisi

d et al. 2005)



2.3 TLC-Bioautography method

TLC-Bioautography is a useful technique to determine bioactive compounds with

antimicrobial activity from plant extracts. TLC bioautographic methods combine

chromatographic separation and in situ activity determination, facilitating the

localization and target-directed isolation of active constituents in a mixture.

Traditionally, bioautographic technique has used the growth inhibition of

microorganisms to detect anti-microbial components of extracts, chromatographed on

a TLC layer. This methodology has been considered as the most efficacious assay for

the detection of anti-microbial compounds (Tradit et al. 2011).

From crude plant extracts, bioactivity oriented isolation of compounds can be

performed by TLC-Bioautography method. TLC-bioautography can be coupled with

several other hyphenated techniques like HPLC, LC-MS, GC-MS and H1 NMR in

order to identify the isolated active constituents. Table 2.3 represents the list of plants

and their isolated compounds by TLC-Bioautography-Hyphenated techniques.

Table 2.3: List of plants and their isolated compounds by TLC-Bioautography-

Hyphenated techniques

Sr.

No

Name of the

Plants Methods Employed

Isolated

compounds References

1

Himanthalia

elongata

TLC bioautography,

UV-visible, FT-IR

Fucoxanthin

(Rajauria &

Abu-Ghannam

2013)

2 Piper betle TLC bioautography,

GC-MS

Eugenol,

Allylpyrocatechol,

eugenyl acetate.

(Annegowda et

al. 2012)

3 Cassia fistula TLC bioautography,

LC-MS Roseanone

(Jothy et al.

2011)

4

Anthocephalus

indicus

TLC bioautography,

FTIR, MS, H1 NMR

Iridoid glucoside (Sanadhya &

Durve 2014)

5

Excoecaria

agallocha

TLC bioautography,

HPLC, H1 NMR

Aliphatic

compounds

(Patra et al.

2012)



2.4 Review of literature of plants selected for the study

Following plants were selected for the study:

Table 2.4: Plants selected for the study

Sr. No Plants Part Selected

1 Ocimum tenuiflorum Linn Leaves

2 Citrus reticulata Blanco Peels

3 Citrus aurantifolia Swingle Peels

4 Butea monosperma Lam Seeds

5 Vitis vinifera Linn Seeds

These plants were selected based on literature review and their traditional use

mentioned in Ayurveda. The literature pertaining to the plants include in the present

study was reviewed with respect to the following points:

A. Classification

B. Synonyms

C. Parts used

D. Botanical description

E. Therapeutic benefits

F. Phytochemical Constituents

G. Pharmacological Activities



2.4.1 Ocimum tenuiflorum Linn

Fig. 2.1: Ocimum tenuiflorum Linn.

A. Classification:-

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnoliopsida

Subclass : Asteridae

Order : Lamiales

Family : Lamiaceae

Genus : Ocimum L.

Species : Ocimum tenuiflorum Linn

B. Synonyms:-

English : Holy Basil, Sacred Basil

Hindi : Tulasi

Marathi : Tulas

Sanskrit : Surasa, Krisnatulasi, Bana Tulasi.

Kannada : Tulasi, Shree Tulasi, Vishnu Tulasi

Telugu : Tulasi

Malayalam : Tulasi, Tulasa

Tamil : Tulasi, Thulasi, Thiru Theezai



C. Parts used:-

Whole plant

D. Botanical description:-

Ocimum tenuiflorum Linn is an annual erect herb, much branched and 30-60 cm

high. Leaves are simple, opposite, elliptic-oblong, margins serrate, hairy along the

vein. Leaves are 2-4 cm in length. Flowers are small. Each flower consist of

pubescent calyx, corolla 2-lipped, stamens 4 in 2 pairs, filament slender,the upper

pair with small appendages at base, style 2-lobed. Fruits are nutlets smooth and

broadly elliptic. Seeds are rounded to oval in shape, brown in color, 0.1 cm long

and slightly notched at the base.

E. Therapeutic benefits (Warrier et al. 2006)

The plant is bitter, aromatic, stomachic, diaphoretic, digestive and diuretic. It is

useful in cardiopathy, haemopathy, leucoderma, asthma, bronchitis, fever,

hepatopathy, vomiting, ophthalmia, gastropathy in children, genital-urinary

disorders, ringworm and skin diseases.

F. Phytochemical constituents:-

The chemical constituents of Ocimum tenuiflorum leaves can be categorized as

volatile and non-volatile compounds. Table 2.5 represents the major

phytoconstituents present in the Ocimum tenuiflorum Linn.



Table 2.5: Major phytoconstituents present in

Ocimum tenuiflorum Linn

Sr.

No Name Structure Reference

1 Eugenol

(Kelm et al. 2000)

2 Carvacol

(Yanishlieva et al.1999)

3 Linalool

(Khan et al. 2010)

4 Caryophylline

(Devendran et al. 2011)

5 Estragol

(Pattanayak et al. 2010)

6 Ursolic acid

(Shishodia et al. 2003)

7 Rosmarinic

acid

(Hakkim et al. 2011)

8 Apigenin

(Dutta et al. 2007)

9 Vanillin

(Dev et al. 2010)

10 Stigmasterol

(Singh et al. 2012)



G. Pharmacological activities:-

Anti-diabetic activity

Leaves of Ocimum tenuiflorum have been traditionally used in treatment of diabetes

mellitus. Sethi et al showed that the dietary supplementation of fresh tulsi leaves in

a dose of 2 gm/ kg body weight for 30 days led to significant lowering of blood

glucose levels in albino rabbits and made an attempt to study the hypoglycemic and

antioxidant effect of Ocimum tenuiflorum leaves and probable link between these

two effects (Sethi 2004).

Oral administration of ethanolic extract of Ocimum tenuiflorum leaves (50%)

exhibited hypoglycemic effects and led to marked lowering of blood sugar level in

normal, glucose fed hyperglycemic and streptozotocin induced diabetic rats

(Chattopadhyay 1993)

The anti-diabetic effects of ethyl acetate, petroleum ether and chloroform fractions

from ethanolic extract of the Tulsi leaves were investigated in normal and alloxan

induced diabetic rats. The hypoglycemic and hypolipidemic activities were studied

by various parameters like fasting blood glucose, total cholesterol, triglyceride,

serum glutamate oxaloacetate transaminases, serum glutamate pyruvate

transaminases level, and liver glycogen content and results indicate that the different

fractions have exhibited favorable effects in bringing down the severity of diabetes

together with hepatoprotectivity (Khan et al. 2009).

Anti- microbial activity

Various studies were conducted to prove the antibacterial and anti-fungal activities

of Ocimum tenuiflorum Linn. Singh et al showed that fixed oil of Ocimum leaves

showed good antibacterial activity against S. aureus, Bacillus pumilus and

Pseudomonas aeruginosa, where S. aureus was the most susceptible organism. This

study suggested that the higher content of linolenic acid fixed oil could contribute

towards its antibacterial activity (Singh et al. 2005).

Ali and Dixit isolated two flavonoid compounds i.e. Orientin, Vicenin from aqueous

extract of fresh leaves and showed their antibacterial activity against bacteria



causing urinary tract infection in human which include Escherichia coli, S.

aureus, Staphylococcus cohni and Klebsialla pneumonia (Ali & Dixit 2012).

Viyoch et al screened the essential oils of various species of Ocimum for

antimicrobial activity against P. acnes. Results indicate that, these essential oils

could be incorporate in suitable formulations for acne skin care (Viyoch et al.

2006).

Prasannabalaji et al evaluated the in vitro antibacterial activity of various solvent

extracts of Indian traditional medicinal plants including Ocimum species.

Antibacterial activity was performed against Escherichia coli, S. aureus, Salmonella

typhi, Salmonella paratyphi and Klebsiella pneumonia and it was concluded that

methanol extracts of Ocimum species showed maximum zone of inhibition against

Salmonella typhi (Prasannabalaji et al. 2012).

Eugenol and Methyl eugenol were identified as the major constituents of essential

oil of Ocimum tenuiflorum Linn. Joshi studied the antibacterial activity of essential

oils of Ocimum, eugenol and methyl eugenol. Numbers of Gram positive, Gram

negative and fungal strains were used as test organisms in the study. Methyl eugenol

exhibited significant activity against Pseudomonas aeruginosa while eugenol was

effective only against S. aureus (Joshi 2013)

Anti- inflammatory activity

Singh and Majumdar studied the anti-inflammatory activity of fixed oil of Ocimum

tenuiflorum. Carrageenan and Arachidonic acid were used to induce inflammation in

Wistar albino rats. Linseed oil and soyabean oil containing linolenic acid were tested

along with fixed oil, showed significant inhibition of carrageenan-induced paw

edema. The results suggest that linolenic acid present in Ocimum tenuiflorum fixed

oil has the capacity to block both the cyclo-oxygenase and lipoxygenase pathways of

arachidonate metabolism and could be responsible for the the anti-inflammatory

activity of the oil (Singh & Majumdar 1997) (Singh 1998) (Singh et al. 2007).



Hannan et al showed that the ethanol extract of Ocimum tenuiflorum exhibited

promising anti-inflammatory effect in rats using carrageenan-induced rat paw edema

model in dose dependant manner. This study supported the use of Ocimum

tenuiflorum as an analgesic and anti-inflammatory drug in folk medicine (Hannan et

al. 2011).

Immunomodulatory activity

Traditionally it is believed that consumption of Tulsi leaves on empty stomach

increases immunity. Mondal et al showed that the 300 mg capsules of ethanolic

extracts of Tulsi leaves have immunomodulatory effect. Double-blinded randomized

controlled cross-over trial was conducted on healthy volunteers and various

parameters like interferon-γ and interleukin-4 cytokines, T-helper and T-cytotoxic

cells, B-cells and NK-cells were analysed (Mondal et al. 2011).

Immunomodulatory activity of aqueous extract of Ocimum tenuiflorum was studied in

Wistar albino rat by Jeba et al. They have administered oral dose of 100, 200

mg/kg/day for 45 days and checked immunomodulatory effect along with the

biochemical and haematological changes. Results showed increasing antibody

production in dose dependent manner (Jeba et al. 2011).

Skin care Activity

Sawarkar et al developed anti-acne topical formulation containing hydro-alcoholic

extract of Ocimum sanctum, ethanolic extract of Tabernaemontana divaricata, Aloe

vera concentrate gel powder and tea tree oil and tested its antibacterial potential

against P. acnes (Sawarkar et al. 2010).

Lertsatitthanakorn investigated the effect of bioactives of essential oil to control acne.

Seven essential oils including holy basil oil were studied against P.acnes. Antioxidant

activity using the DPPH free radical scavenging assay showed that the IC50 values of

holy basil oil were lower than that of ascorbic acid (Lertsatitthanakorn et al. 2006).

Viyoch et al reported the antibacterial activity of Thai basil oils and their micro-

emulsions, against P. acnes (Viyoch et al. 2006).



2.4.2 Citrus reticulata Blanco

Fig. 2.2: Citrus reticulata Blanco

A. Classification:-

Kingdom : Plantae



Subclass : Rosidae

Order : Sapindales

Family : Rutaceae

Genus : Citrus L.

Species : Citrus reticulata Blanco

B. Synonyms:-

English : Loose-skinned Orange, Mandarin Orange

Hindi : Narangi, Sangtara

Marathi : Naringa, Santra

Sanskrit : Naranga, Tvaksugandha

Kannada : Kittale, Herale

Telugu : Kamalepandu, Malli

Malayalam : Madhuranaranna

Tamil : Kamala, Koorg kudagu orang



C. Parts used:-

Leaf, flowers, fruit, rind


The plant is an evergreen bushy moderate sized tree with greenish white glabrous

branches; leaves are slender and unifoliate. Leaflets are elliptic or ovate and acute

or acuminate. Petioles are short and almost wingless. Flowers are white and

fragrant. Fruits are globose or subglobose with thin rind. Rind is easily separating

from the segments and becomes bright orange in color on ripening. Seeds are

many and white in color.

E. Therapeutic benefits (Warrier et al. 2006):-

The fruits are sour, sweet, cooling deodorant, aphrodisiac, digestive, liver tonic and

are useful in vomiting, retching, seminal weakness, anorexia, dyspepsia,

hepatopathy and cardiac disorder. The fruit rind is anthelmintic, antidiarrhoeal,

stomachic and tonic. It is useful in vomiting, helminthiasis, diarrhoea, dyspepsia,

flatulence, skin diseases and general debility.




Table 2.6: Major phytoconstituents present in Citrus reticulata Blanco

Sr.


1 D-limonene

(Chutia et al. 2009)

2 Geranial

(Danielski et al.

2008)

4

Geranyl acetate

(Nautiyal & Tiwari

2014)

5 Linalool

(Danielski et al.

2008)

6

Nerol

(Tailor & Singh

2011)

7 β-Myrcene

(Yu et al. 2009)

8 Hesperidin

(Sun et al. 2010)

9 Tangeretin

(Jasim 2012)

10 Nobiletin

(Jasim 2012)




Antimicrobial activity

Sultana et al investigated the antimicrobial activity of volatile constituents of fruit

peels of Citrus reticulata Blanco against clinically isolated pathogenic

microorganisms. S. aureus was found to be most susceptible organism. Anti-

fungal activity of essential oil was also studied against Aspergillus Niger,

Aspergillus fumigatus, Aspergillus flavus and Candida albicans. Results indicate

the potential antimicrobial activity of volatile oil present in peels of Citrus

reticulata which can be useful for treatment of skin disorder and in aroma therapy.

It can be incorporated into cosmetic formulations (Sultana et al. 2012).

Jayaprakasha et al has performed the successive extraction of Citrus peels with

hexane, chloroform and acetone by Soxhlet extraction. The hexane and chloroform

extracts were further fractionated into alcohol-soluble and alcohol-insoluble

fractions and tested against different gram positive and gram negative bacteria. It

was found that ethanol soluble fraction was most effective against test bacteria and

this activity could be due to isolated polymethoxylated flavones from same

fraction. Characterisation of these isolated flavones was performed by UV, 1H,

13C NMR and mass spectral studies. The compounds identified were

desmethylnobiletin, nobiletin and tangeretin (Jayaprakasha et al. 2000).

The antifungal activity of Citrus peel essential oil was tested against five plant

pathogenic fungi viz Alternaria alternate, Rhizoctonia solani, Curvularia lunata,

Fusarium oxysporum and Helminthosporium oryzae. Essential oils obtained by

hydro-distillation method was analysed by Gas Chromatography (GC) and Gas

Chromatography–Mass Spectroscopy (GC-MS) (Chutia et al. 2009).

Anti-inflammatory activity

Ho et al have isolated and identified hydroxylated polymethoxyflavones from

citrus peels and investigated their anti-inflammatory and cancer chemopreventive

property. The outcome of the study suggest that, hydroxylated

polymethoxyflavones possess greater biological activity potency for

chemoprevention on targeting inflammation (Ho et al. 2012).



Ho and Lin determined the anti-inflammatory activities of heat-treated Citrus peel

extracts. Results showed that the anti-inflammatory activity of Citrus peel was

significantly elevated after 100 ºC heat treatment in a time-dependent manner

during a period from 0 to 120 min. The anti-inflammatory activity of Citrus peel

extract was correlated with the content of nobiletin and tangeretin and it was

concluded that, the proper and reasonable heat treatment helped to release nobiletin

and tangeretin, which were responsible for the increased anti-inflammatory activity

of heat-treated Citrus peels (Ho & Lin 2008).

Huang and Ho demonstrated, for the first time, that polymethoxy flavones,

especially nobiletin, were responsible for the inhibitory activity of Citrus fruit

peels on prostaglandin E2 (PGE2) production. They have selected seven Citrus

fruits for this study and determined the inhibitory ability of Citrus peel extracts on

the production of pro-inflammatory mediators. Among all, peels of Citrus

reticulata Blanco and Citrus tankan Hayata showed promising results and anti-

inflammatory activity (Huang & Ho 2010).

Antioxidant and Cardiovascular activity:

Vinson et al studied the effect of polyphenol antioxidants in Citrus juices and their

correlation with heart disease through in vitro and in vivo models. Polyphenol

quality in the juices was analyzed by using the inhibition of lower density

lipoprotein oxidation promoted by cupric ion, an in vitro model of heart disease. In

a hamster model of atherosclerosis, the juices were able to significantly inhibit

atherosclerosis and lowered cholesterol and triglycerides (Vinson et al. 2002).

Rincón et al showed that the Citrus reticulata peel has highest anti-radical activity

and it was correlated with the polyphenol content. The results of this study suggest

that the Citrus peel should be the most suitable, to reduce risk of cardiovascular

diseases because the studied samples were good sources of dietary fiber and

phenolic compounds (Rincón et al. 2005).



Anti-cancer activity

Gao et al showed that the Citrus flavonoid naringenin stimulates DNA repair in

LNCaP human prostate cancer cells. Results demonstrated that the naringeni

stimulate DNA base excision repair and prevent mutagenic changes in prostate cancer

cells (Gao et al. 2006).

Du and Chen identified the four major polymethoxyflavones viz. isosinensetin,

sinensetin, nobiletin and tetramethyl-o-scutellarein and studied their anti-proliferative

activity against cancer cell lines (A549, HL-60, MCF-7 and HO8910) and showed

that isosinensetin was most effective among all. Results suggested that the Citrus

peels are excellent sources of functional polymethoxyflavones that may help prevent

female cancers, such as ovarian cancer and breast cancer (Du & Chen 2010).

Skin care Activity

Wang et al studied the cosmetic applications of selected traditional Chinese herbal

medicines including Citrus reticulata Blanco peel. Authors have tested for

cytotoxicity on human epidermal melanocytes and evaluated their effects on

tyrosinase and melanin inhibitory activities. Free radical scavenging activities and

total phenolic content were also determined (Wang et al. 2006).



2.4.3. Citrus aurantifolia (Christm) Swingle

Fig. 2.3: Citrus aurantifolia (Christm) Swingle

A. Classification:-

Kingdom : Plantae



Subclass : Rosidae

Order : Sapindales

Family : Rutaceae

Genus : Citrus L.

Species : Citrus aurantifolia (Christm) Swingle

B. Synonyms:-

English : Acid Lime, Sour Lime

Hindi : Kaghzi nimbu

Marathi : Limbu

Sanskrit : Jambhiir

Kannada : Limbe, Nimbe

Telugu : Nimma

Malayalam : Vatukappulinarakam

Tamil : Elumichai



C. Parts used:-

Fruits, Rind, Leaves.


A medium sized thorny tree or a shrub with greenish white, glabrous young shoots and

greyish brown bark; leaves foliolate, leaf-stalks broadly winged, leaflet are elliptic or

ovate, acute, obtuse; flowers are white, large, very fragrant; fruit globose, bright

yellow when ripe, rind of fruit is very aromatic, pulp sour, bitter; seeds many, yellow

or coloured, smooth and slimy.

E. Therapeutic benefits (Warrier et al. 2006b) :-

The fruits are sour, bitter, astringent, thermogenic, laxative, stomachic, digestive,

anthelmintic and antiscorbutic, and are useful in vitiated conditions of Pitta and

Kapha, bronchitis, dyspepsia, nausea, flatulence, colic, helminthiasis, scabies and

anaemia.




Table 2.7: Major phytoconstituents present in

Citrus aurantifolia (Christm) Swingle

Sr.


1 Limonene

(Spadaro et al.

2012)

2 α -Pinene

(Yadav et al.

2004)

3

β-Pinene

(Yadav et al.

2004)

4 α -terpineol

(Arias & Ramón-

Laca 2005)

5 Linalool

(Lota et al. 2002)

6 Bergamottin

(Sandoval-

Montemayor et al.

2012)

7 Myrcene

(Sakanaka &

Cabral 2006)

8 γ-terpinene

(Dugo et al. 1997)





Aibinu et al investigated the antimicrobial activity of different parts of Citrus

aurantifolia as against various clinical isolates namely S. aureus, Salmonella

paratyphi, Shigella flexnerii, Streptococcus faecalis, Citrobacter spp, Serratia spp,

Klebsiella pneumoniae, Pseudomonas areuginosa etc. Anti-fungal activity was

studied and it was concluded that peel oil and epa-ijebu (burnt rind of the fruit) shown

maximum antibacterial effect (Aibinu et al. 2006).

Chanthaphon et al studied the antimicrobial activities of essential oils and crude

extracts from tropical Citrus species against food-related microorganisms and showed

that the ethyl acetate extract from lime peel showed broad spectrum inhibitory activity

against all Gram-positive bacteria tested (Chanthaphon et al. 2008).

Pathan et al evaluated the in vitro antimicrobial activity of Citrus aurantifolia leaf

extracts prepared in differert solvents. Results demonstrated that the hydroalcoholic

extract was found to be more potent against S. aureus, Escherichia coli, Klebsiella

pneumonia, Pseudomonas spp., Aspergillus niger, Aspergillus fumigates, Mucor spp

(Pathan et al. 2012).

Anti-cholinesterase activity

Tundis et al performed the comparative study on the antioxidant capacity and

cholinesterase inhibitory activity of various Citrus peel essential oils and shown that

Citrus aurantifolia oil showed the highest radical scavenging activity on ABTS assay

(IC₅₀ value of 19.6 μg/ ml and inhibited the acetylcholinesterase more selectively.

The study suggests a potential use of Citrus oils as a valuable new flavor with

functional properties for food or nutraceutical products with particular relevance to

supplements for the elder people (Tundis et al. 2012).

Loizzo et al evaluated the antioxidant and anti-cholinesterase activity of Citrus

aurantifolia peel and leaves extracts and concluded that the methanol extracts of the

peel and leaves demonstrated the strongest radical scavenging activity where as



n-Hexane fractions of both peel and leaves showed a good acetylcholinesterase

inhibitory activity (Loizzo et al. 2012).

Anti- atherosclerotic activity

Boshtam et al evaluated the effects of Citrus aurantifolia juice and peel on

antioxidant activity and atherosclerosis progression in rabbits receiving a hyper-

cholesterolemic diet. They have analyzed the serum lipids and measured the

antioxidant activity. The rabbits‟ aorta and coronary arteries were separated and the

presence of fatty streaks was studied. Results showed that, peel and juice increase

plasma antioxidant capacity in rabbits, and can thus prevent or decelerate the process

of atherogenesis. However, lime peel was more effective than lime juice (Boshtam et

al. 2013) (Boshtam et al. 2011) .

Antitumor activity

Gharagozloo1 et al investigated the anti-proliferative effect of concentrated lime juice

on the human breast carcinoma cell line (MDA-MB-453) and human lymphoblastoid

B cell line (RPMI- 8866). It was found that, this extract had no significant effect on

MDA-MB-453 cell line; however, significant inhibition of the spontaneous

proliferation of RPMI-8866 cell line was detected at 125, 250, and 500 µg/ ml of

concentrations. Authors have corelated the anti-proliferative activity of test extract

with protein nature of the biologically active macromolecules present in the extract

(Gharagozloo et al. 2002)

Principle components of volatile oil of Citrus aurantifolia peel were identified by

Patil et al and 78% inhibition of human colon cancer cells (SW-480) was shown at

100 µg/ ml concentration after 48 hrs. Results demonstrated the principles of volatile

oil can induce apoptosis-mediated cells death in human colon adenocarcinoma cells

(Patil et al. 2009)



2.4.4. Butea monosperma Lam.

Fig. 2.4: Butea monosperma Lam.

A. Classification:-

Kingdom : Plantae



Subclass : Rosidae

Order : Fabales

Family : Fabaceae

Genus : Butea Roxb. ex Willd

Species : Butea monosperma (Lam.) Taubert

B. Synonyms:-

English : Bengal Kinotree

Hindi : Dhak, Palash

Marathi : Palash

Sanskrit : Kinsuka, Raktapuspaka, Ksara srestha, Brahma Vrksa

Kannada : Muttuga

Telugu : Moduga mada

Malayalam : Palashu

Tamil : Purashu



C. Parts used: -

Bark, leaves, flowers, fruits, seeds, gum.


A medium sized deciduous tree, very conspicuous when in flower, 12-15 meter in

height, trunk is somewhat crooked, leaves are 3-foliate and leaflets are obtuse,

glabrous above when old, silky and reticulately veined beneath. Flowers are bright

orange red, fruit pods are thickened at the sutures, containing a single seed.


The bark is bitter, astringent, emollient, aphrodisiac, appetizer, digestive,

anthelmintic and tonic, useful in vitiated conditions of Pitta and Kapha, useful in

dyspepsia, cooling, constipating, aphrodisiac, diarrhoea, intestinal worms and

bone fractures. The leaves are astringent and anti-inflammatory. The flowers are

astringent, sweet and cooling, diuretic and tonic. Seeds are useful in Pama,

Kandu, Dadru, Tvak dosha, nashaka – usfeul in allergic dermatitis, pruritis,

contact dermatitis etc. It helps to detoxify skin.




Table 2.8: Major phytoconstituents present in Butea monosperma Lam.

Sr. No Name Structure Reference

1 Palasonin

(Kaleysaraj &

Kurup 1967)

2 Linoleic acid

(Prasad et al. 1987)

3

Palmitic acid

(Prasad et al. 1987)

4 n-docosanoic acid

(Alam et al. 2010)

5

2 – Hydroxy – 1 –

methyl allophonic

acid

(Porwal et al. 1988)

6 15 – hydroxyl

pentacosanoic acid

(Sharma et al.

1991)

7 Monospermine

(Mehta & Bokadia

1981)

8 Palsonin – N –

phenyl imide

(Guha et al. 1990)




Anthelmintic activity

Singh et al investigated the anthelmintic efficacy of aqueous extract of Butea

monosperma seeds against Haemonchus contortus. The seed extract showed

complete mortality of worms at the concentration of 100 mg/ ml at the time exposure

of 6 hrs and with the concentration of 50 mg/ ml at the post exposure of 8 hrs.

Authors concluded that these cidal effects colud be due to presence of high phenolic,

flavonoids and tannin content in the extract (Singh et al. 2013).

The crude extracts of Butea monosperma leaves were investigated for their

anthelmintic activity against earthworms (Pheretima posthuma), roundworms

(Ascardia galli.) and tapeworms (Raillietina spiralis). Alcohol and ethyl acetate

extracts exhibited significant anthelmintic activity. Albendazole was used as a

standard reference in the study (Borkar et al. 2010).

Iqbal et al studied the in vivo anthelmintic activity of Butea monosperma against

Trichostrongylid nematodes in sheep and showed a dose-dependent (1–3 g/ kg) and a

time-dependent anthelmintic activity in sheep (Iqbal et al. 2006).


Ahmad and Khan studied the antibacterial activity of leaves, bark, flowers and seeds

extract of Butea monosperma against S. aureus, Klebsiella pneumonia, Pseudomonas

aeruginosa, Bacillus subtilis, Escherichia coli. Results demonstrated that, the

methanol and aqueous extracts of different parts of plants exhibited comparatively

higher antibacterial activity (Ahmad & Khan 2012).

The antimicrobial activity of Butea monosperma leaf extracts was evaluated by the

agar-well diffusion method against clinically isolated 12 Gram-positive and -negative

multidrug resistant pathogenic bacteria. Sahu and Padhy concluded that, the leaf-

extracts with hot water and ethanol had shown significant antibacterial activity against

all bacteria and the extract could be used in treating infectious diseases, caused by the

range of tested bacteria, as complementary and alternative medicine

(Sahu & Padhy 2013).



Anti-diabetic activity

Anti-diabetic potential of ethanolic extract of Butea monosperma flowers was studied

in glucose-loaded and alloxan-induced diabetic rats and it was shown that repeated

oral treatment of extract (200 mg/ kg/ day) for 2 weeks significantly reduced blood

glucose, serum cholesterol and improved HDL-cholesterol and albumin as compared

to diabetic control group (Somani et al. 2006).

Bavarva and Narasimhacharya studied the antihyperglycemic and antihyperlipidemic

effects of ethanolic extract of Butea monosperma in non-insulin dependent diabetes

mellitus rats and found that the dose of 300 mg/ kg body weight exhibited significant

antidiabetic, hypolipidemic and antiperoxidative effects (Bavarva & Narasimhacharya

2008).



2.4.5. Vitis vinifera Linn

Fig. 2.5: Vitis vinifera Linn

A. Classification:-

Kingdom : Plantae



Subclass : Rosidae

Order : Rhamnales

Family : Vitaceae

Genus : Vitis L.

Species : Vitis vinifera Linn

B. Synonyms:-

English : Grapes

Hindi : Angur

Marathi : Draksha

Sanskrit : Draksha

Kannada : Draksha, Angura

Telugu : Draksha kottai, Drakshai

Malayalam : Munthringya, Buaangur

Tamil : Kotumuntiri



C. Parts used:-

Ripe fruit (dried), leaf, stem, flower, seeds


A large, perennial tendril climber; tendrils leaf opposed, often bifid. Leaves

simple, rotund-cordate or orbicular-cordate, dentate, 3-7 lobed, 10-12 cm across,

glabrous above, tomentose beneath. Flowers in long peduncled, leaf- opposed

cymes, greenish or white. Fruits (berry) globose, ovoid or oblong, varying in size,

pale green or purple. Seeds 2-4, oblong-obovoid, brown, with a discoidal tubercle

on the back.


The fruits are sweet, refrigerant, laxative, demulcent, intellect promoting,

cardiotonic, haematinic, haemostatic, diuretic, aphrodisiac, rejuvenating, nervine

tonic, febrifuge, depurative, antispasmodic, digestive, stomachic, suppurative,

expectorant and tonic. They are useful in burning sensation, constipation, amentia,

cardiac debility, haemoptysis, haemorrhages, anaemia, fever, leprosy, skin diseases,

dyspepsia, colic, flatulence, cough, asthma, bronchitis, infections of eyes and throat,

gout, jaundice and general debility. The seeds are useful in skin diseases.




Table 2.9: Major phytoconstituents present in Vitis vinifera Linn

Sr. No Name Structure Reference

1 Proanthocyanidins

(Saito et al. 1998)

2 (+)-Catechin

(Georgiev et al. 2014)

3 (−)-epicatechin

(Georgiev et al. 2014)

4

Epigallocatechin

(Guendez 2005)

5 Epicatechin-3-O-

gallate

(Jayaprakasha et al.

2003a)

6 Gallic acid

(Prodanov et al. 2013)




Anti-ulcer activity

Saito et al reported the antiulcer activity of grape seed extract and procyanidins in

rats. Grape seed extracts with low and high flavonols content and procyanidins

strongly inhibited the stomach mucosal injury at a dose of 200 mg/ kg (Saito et al.

1998).

Wang et al investigated the therapeutic effect and mechanism of proanthocyanidins

from grape seed in the treatment of recurrent ulcerative colitis in rats.

Proanthocyanidin rich grape seed extract exerted a protective effect on recurrent

colitis by modifying the inflammatory response and inhibiting inflammatory cell

infiltration. It also promoted the damaged tissue repair by inhibiting the nitric oxide

synthase activities (Wang et al. 2010). The authors further studied the molecular

mechanism involved in the therapeutic effects of proanthocyanidins from grape seeds

on recurrent ulcerative colitis in rats and demonstrated that the expression levels of

tumor necrosis factor-α , inhibitor kappa B kinase , protein levels of inhibitory kappa

B-alpha and the translocation of nuclear translocation levels of nuclear factor-kappa B

were significantly reduced in the colon mucosa (Wang et al. 2011).


Jayaprakasha et al studied the antibacterial and antioxidant activities of grape seed

extracts. Antibacterial activity was tested against Bacillus cereus, Bacillus coagulans,

Bacillus subtilis, S. aureus, Escherichia coli and Pseudomonas aeruginosa and it was

found that, lower concentration was required for inhibition of Gram-positive bacteria

than Gram-negative bacteria (Jayaprakasha et al. 2003)

Baydar et al determined the total phenolic content and antimicrobial activity of grape

seed and bagasse extracts. Antimicrobial activity was checked against some food

spoilage and pathogenic bacteria and it was found that the grape seed extracts at 4%

and 20% may be useful as antibacterial agents to prevent the deterioration of food

products (Baydar et al. 2004).



Skin-care Activity

Extensive research suggests that grape seed extract is beneficial in many areas of

health because of its antioxidant effect to bond with collagen, promoting youthful

skin, cell health, elasticity, and flexibility and proanthocyanidins are responsible for

protecting body from sun damage (Yamakoshi et al. 2004)

Cornacchione et al evaluated the antioxidant effect of Vitis vinifera shoot extract on

normal human keratinocytes and the extract exhibited stronger activity than vitamin C

and vitamin E. Authors have compared the extract in combination with a

biotechnological extract (Ronacare Hydroine), and evaluated the efficacy on

photoaging skin. Results indicate that a 4-week twice-daily application of a serum

containing the combination improved the main clinical signs of photoaged skin

(Cornacchione et al. 2007).

Ravichandran and Kolhapure evaluated the clinical efficacy and safety of anti-wrinkle

cream in the management of facial skin wrinkles. Anti-wrinkle cream was a

polyherbal formulation including Vitis vinifera as one of the component. The principle

constituents of Vitis vinifera are flavonoids and glycosylated stilbenes which have

strong antioxidant potential and could be responsible for anti-wrinkle effect

(Ravichandran & Kolhapure 2005).

Yamakoshi et al studied the effect of oral intake of proanthocyanidin-rich extract of

grape seeds on chloasma. The results demonstrated that grape seed extract is a

powerful antioxidant and capable is reducing the facial hyperpigmentation of

Japanese women with chloasma in a one-year study (Yamakoshi et al. 2004).

Grape seed oil is useful in minimizing the skin aging due to its antioxidant properties.

Fine lines and wrinkles are common indications of aging processes, but the oil may

help to reduce the appearance of those signs by providing moisture and enough

protections against free radicals (Anon n.d.).

Documents

REVIEW OF LITERATURE - Shodhgangashodhganga.inflibnet.ac.in/bitstream/10603/73024/9/08. chapter 2.pdf · linked to skin aging and is co-related with an increase in activity of certain