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Amir Hassan, HimayatUllah, Muhammad Israr
Page | 732
The antioxidant activity and phytochemical analysis of medicinal plant Veronica biloba
Amir Hassan1, *
, Himayat Ullah1
, Muhammad Israr2
1Department of Organic Chemistry; Government Post Graduate College Mardan (23200), Affiliated with Abdul Wali Khan University Mardan Khyber Pakhtunkhwa
Pakistan. 2Department of Botany; Government Post Graduate College Mardan (23200), Affiliated with Abdul Wali Khan University Mardan Khyber Pakhtunkhwa Pakistan.
*corresponding author e-mail address:[email protected] | Scopus Author ID: 57211903251
ABSTRACT
A medicinal plant veronica genus has 450 well known species and found across both temperate and hemisphere region their 26 species are endemic and
known in a total of 79 popular species and are widely utilized throughout the world due to important biological activities. In this study fully powdered
uniform size specie veronica biloba plant taken in porous bag were manually subjected to soxhlet hot continuous process for cyclization of extraction
using ethanol (300 mL) a concentrated dried extract obtained after solvent evaporation. Furthermore, liquid-liquid extracted fractions as water,
dichloromethane, n-hexane, and ethyl acetate yields results found polar fraction with the highest percentage (water 47.51 %). The phytochemical
screening of veronica biloba has shown all major compounds entirely present in extracts. One of the primitive phenolic compound flavonoid is present
in plant and show potency towards antioxidants. All the extracted fractions of plant showed excellent antioxidant activity using a stable DPPH (1,1-
diphenyl-2-picryl-hydrazyl) method at concentration (range from 31.25 to 500µg/mL). Primary our ethyl acetate extract fraction showed the highest
inhibition potential at IC50 = 1.70±0.05µg/mL which is much closer to a standard positive control Propyl gallate showed IC50 = 1.6±0.05µg/mL percent
potential. The purification and isolation of these extract areimportant which can provide us help in novel antioxidants discovery also natural antioxidants
currently in cosmetics products, food and therapeutics health related products significantly demanded because they are very effective, efficient and
harmless as compared to synthetic.
Keywords: Soxhlet; Phytochemicals; DPPH; Antioxidants; Percent Yields.
1. INTRODUCTION
A medicinal plant veronica genus has 450 well known
species and found across both temperate and hemisphere region
[1]. In veronica plant their 26 species are endemic and known in a
total of 79 popular species [2]. Veronica species are widely
utilized throughout the world due to higher chemotaxonomic,
phytochemicals value, and important biological activities
investigated on it therefore mostly utilized by Chinese and Turkish
also in conventional or, traditional medicinal system for tonics,
wound healing, influenza, expectorants, restoratives, and also for
respiratory infection [3]. Some species of this plant areutilized in
anticancer treatment [4]. Extractions from medicinal plants
possess certain potentially active substances or, compounds which
is accountable for biological activity [5]. The most recent study
investigated in a medicinal plant founds mainly antioxidants, such
as flavonoids, Vitamin A, C, E, Lignin’s, and tannins phenolic
compounds are antioxidants and exist in plants [6]. The plants
extracted natural products give help in the new development of
pure or, standardized compounds in chemical diversity and
unknown availability [7]. The natural product is very effective,
efficient and has no or, little side effects as compared to synthetic
drugs which have side effects and is therefore people utilized
conventional drug obtained from plants [8]. The medicinal plants
possess several novels biological like antioxidants, antiviral,
antitumor, antimalarial, and antimicrobial activities and necessary
to examine these plants and are compulsory to extract the potent
compound may act as natural antioxidants or, in other activities
instead of synthetic one [9]. Currently in cosmetics products, food
and therapeutics health related products significantly demanded
the natural antioxidants having multifacetedness high amount and
mass [10]. Muchnoticeable biological activity reported on the
veronica plant species like antibacterial [11]. Mocan et al [12]
found in veronica orchidea, officinallis, teucrium, antioxidants,
phytochemical and antimicrobial. Dunkic et al [13] in veronica
spicataconfirmed antioxidants plus antimicrobial while Exarchou
[14] isolated compounds hispidulin also antiviral [15] an effective
anti-cancer, anti-mutagenic and antitumor with new compounds
[16-19] anti-ulcerogenic [20, 21] anti-cholinesterase [22] anti-
inflammatory [18] anti-insecticidal [23] anti-hepatocarcinoma [24]
antioxidants [25] angiogenic, neuroprotective potency [26] wound
healing, cough, catarrh, and kidney medication [12] natural
preservatives, dietary supplement, and hyperglycemia [27, 28] was
reported in this genus species, Javadsharif rad et al [29] found in
veronica persicapoiracetylcholinesterase, scolicidal, tyrosinase,
xanthine oxidase, lipoxygenase. Our current study is to focused
only on antioxidant analysis by (1,1-diphenyl-2-picryl-hydrazyl)
method and phytochemical confirmation also yield calculation for
the first time in the genus veronica medicinal plant a specie
veronica biloba.
2. MATERIALS AND METHODS
2.1. Material and Methods.
2.1.1. Collection and identification of the plant.
The medicinal plant veronica biloba confirmed and indentified via
a literature-survey also from Prof. Muhammad Israr botanical
export of Govt: Post Graduate College Mardan. The healthy and
fresh medicinal plant utilized in the project-experiment, plant at
flowering stage was collected during February-March months.
Selection is done from specific fertile land location Par HotiSang-
e-mar mar and RustamSurkhDheri tehsil and district 23200
Mardan. A Collected plant with roots washed with sterilized
Volume 8, Issue 4, 2019, 732 - 738 ISSN 2284-6808
Open Access Journal Received: 19.11.2019 / Revised: 20.12.2019 / Accepted: 23.12.2019 / Published on-line: 27.12.2019
Original Research Article
Letters in Applied NanoBioScience https://nanobioletters.com/
https://doi.org/10.33263/LIANBS84.732738
The antioxidant activity and phytochemical analysis of medicinal plant Veronica biloba
Page | 733
water and stabilized through air-exposure in cover and all dust
contamination along with water is removed.
2.1.2. Grinding and drying of the plant.
All the collected plant kept in a shade and avoided from light and
environmental-contamination exposure for three weeks (3-weeks).
After drying process converted into smaller pieces then equal size
powder achieved with extensivesurface area after grinding process
for a better process of extraction.
2.1.3. Soxhlet Extraction Process.
Finally uniform size fully powdered grinded 30 gram plant taken
in porous bag manually prepared from cellulose hard filter-paper
then placed in Soxhlet thimble section. In bottom flask of Soxhlet
300 mL absolute ethanol taken for cyclization of extraction
process. To condenser section water inlet-outlet wasprovided. The
mantox heater temperature maintained at 40 – 450C. The cycling
of solvent occurs through siphon arm up to 36 hrs until the
evaporated solvent does not bring any extracted residue. The dried
extract obtained after ethanol evaporation over the electrical
controllable water bath. Finally, fractions extract achieved from
after liquid-liquid extraction such as water, dichloromethane, n-
hexane, and ethyl acetate get concentrated for phytochemical
screening and for various biological analyses.
2.1.4. Phytochemicals Confirmation analysis.
The medicinal plants constitute several important biologically
active classes of compounds known as phytochemicals. Certain,
plants possess this type of characteristics the identification and
confirmation arevery useful and several studies assigned the
standard protocol for it we have used the procedure of previously
reported in [30-33] discussed given below:
2.1.5. Carbohydrates Confirmation.
Molish Test: The aqueous 3 mL freshly prepared extract taken in
test tube followed by 0.5 mL addition of Molish reagent and
slightly 2-3 drops concentrated H2SO4 added to test tube walls a
violet color ring appeared confirmed the carbohydrates presence.
2.1.6. Tannins Confirmation.
The aqueous 3 mL freshly prepared extract taken in test tube
followed by 0.5 mL addition of Ferric Chloride (FeCl3) solution a
condensed green color confirmed the tannins presence.
2.1.7. Saponins Confirmation.
Froth Test: The aqueous 3 mL freshly prepared extract taken in
test tube shaken vigorously by hand about 3 cm froth is formed in
test tube for about 5-15 minute confirmed the saponins presence.
2.1.8. Flavonoids Confirmation.
The aqueous 3 mL freshly prepared hydrochloric acid extract
taken in two test tubes one followed by 1.5 mL distilled water and
other by 1.5 mL sodium hydroxide (NaOH) addition compare both
with one another a yellow color achieved confirmed the flavonoids
presence. Another method 3 mL extract treated with 1.5 mL of
lead acetate solution a yellowish precipitate obtained positive
result indicates (flavonoids).
2.1.9. Sterols and Triterpenes Confirmation.
Lieberman’s Test: The 3 mL (EtOH) freshly prepared extract
taken in test tube followed by 0.5 mL addition of acetic anhydride
slightly 2-3 drop concentrated H2SO4 added to test tube wall a
brown-reddish color ring appear in upper layer confirmed the
sterol and triterpenes test. Salkowski Test: The 3 mL (EtOH)
freshly prepared extract taken in test tube followed by 2-3 drop
addition of concentrated H2SO4 slightly to the wall of test tube a
reddish brown color ring without greenish layer confirmed the
sterol and triterpenes presence.
2.1.10. Anthraquinone Glycosides Confirmation.
The 3 mL (EtOH) freshly prepared extract taken in test tube
followed by 1 mL addition of ammonia solution a green color in
combination with light rose color confirmed the anthraquinone
glycosides presence.
2.1.11. Resin Confirmation.
The aqueous 4.5 mL extract wasmixed with 2 mL acetone-water
of (1:1) no formation of turbidity seen in solution confirmed the
resin absence.
2.1.12. Protein and amino acid Confirmation.
Xanthoproteic Test: 3 mL extract in test tube followed by 2-3 drop
addition of concentrated nitric acid (HNO3) a yellow color
achieved confirmed the protein presence. Biuret Test: 4.5 mL
extract in test tube addition 1 mL NaOH (10 %) solution then
heated, and consequently added 0.5 mL copper-sulphate (CuSO4)
10 % solution confirmed the protein presence.
2.1.13. Tannins Confirmation.
4.5 mL extract wastreated with 1.5 mL Gelatin (1%) solution in
NaCl combination a white color precipitate achieved confirmed
the tannins presence.
2.1.14. Fats and Fixed Oils Confirmation.
Aqueous extract approximately 2.5 mL pressed between the filter
papers no seen of any oily stain confirmed the absence of fats and
fixed oils.
2.1.15. Diterpenes Confirmation.
3 mL aqueous extract in test tube wasfollowed by 2-3 drop
copper-acetate solution a bright green color obtained confirmed
presence of diterpenes.
2.1.16. Alkaloids Confirmation.
3.5 mL freshly prepared to dilute hydrochloric acidic extract
wasfollowed by 0.5 mL reagent addition of (Mayer’s, Hager and
Wagner separately added) a formation of color precipitate
indicated alkaloids presence.
2.2. Antioxidant Activity by DPPH Method.
All the dried concentrated extracted fractions (water, ethyl-
acetate, dichloromethane, and n-hexane) obtained were evaluated
for antioxidant activity using stable,DPPH (1,1-diphenyl-2-picryl-
hydrazyl) radicals scavenging power effect according to previous
[34-36]. In first all the fractions extracts dissolve in suitable
solvent taking 10 mg/mL in Eppendrops tubes and diluted by ½ in
a serial to (31.25, 62.5, 125, 250, and 500) µg/mL. The standard
DPPH (1, 1-diphenyl-2-picryl-hydrazyl) solution was made 2 hrs
prior tomethanol (100 mL) from 0.0238 gram by weight and store
in dark at room temperature covered with aluminum foil to avoid
from decomposition. In second stage the serial dilution (ranging
from 500 – 31.25 µg/mL) of each extracted fractions were mixed
separately using 10 µL of a single serial with 90 µL DPPH
solution and incubated for 30 minute at room temperature (370C),
at 517 nm absorbance was measured also in triplicates. In the
same concentration (from 31.25 to 500 µg/mL) positive controls
PG (Propyl Gallate), TBH (3-Tert-butyl-4-hydroxyanisol), and
AA (ascorbic acid) were prepared, and placed DMSO
(dimethylsulfoxide) as a negative control media. In last the percent
inhibition of free radical DPPH scavenging activity is measured
according to [37] equation.
Amir Hassan, Himayat Ullah, Muhammad Israr
Page | 734
Antioxidant activity percent % inhibition
=
Whereas Control Abs = absorbance of standard solution, and
Sample Abs = absorbance of the sample used the assay
experiments were repeated in triplicates and median inhibitory
concentration calculated using regression statistics and standard
deviation taking concentration against percent scavenging
inhibition.
3. RESULTS
3.1. Fraction Extracts Yields Calculation.
The percent yield by calculated according to [31] net dried extract
by weight obtained from after Soxhlet hot continuous extraction
process was 7.8314 (gram) about 26.10% by subjecting 30 (gram)
powdered plant to processing. Furthermore, liquid-liquid extracted
fractions yields results almost varied from one another in each
solvent. Fractions percent taken from concentrated extract
following sequence in increasing order as shown water >n-hexane
> ethyl-acetate > dichloromethane variation in percent yields are
water = 47.8%, n-hexane = 28.74%, ethyl-acetate = 20.45% and
dichloromethane = 3.28% shown in [Table 1] also in reported
literature [10, 38-42] results which is based on compounds in
plants by polarities [31] is comparable to our data and we have
found polar fraction with excellent percent yields as shown in
[Figure 1].
Figure 1. Percent-yields comparison of each concentrated fraction.
Table 1. Calculated yields values of veronica biloba for fractions extracts.
S.NO Solvent Fraction Extracts Yield Values
1 Ethyl-acetate Fraction 20.45 %
2 Dichloromethane Fraction 3.28 %
3 n-hexane Fraction 28.74 %
4 Water Fraction 47.51 %
3.2. Phytochemical Screening Confirmation.
Phytochemical screening of medicinal plant veronica
biloba evaluated according to [30-33] almost all major compounds
entirely present in extracts such as flavonoids, tannins, terpenoids,
phenol, alkaloids, protein and amino acids, glycosides and
anthraquinine, carbohydrates, saponins and sterol except fats and
oil and resin test found negative results as shown [Table 2] One of
the primitive phenolic compounds is flavonoids their presence in
plant show potency towards antioxidants [43] while steroids
shows inflammatory potential [44] and alkaloids mostly towards
antispasmodic, analgesic and antimicrobial [43-45].
Table 2. Phytochemical screening results of Veronica Biloba.
S. No Tested Phytochemical Positive
Appearance
Negative
Appearance
1 Alkaloids Positive ----
2 Flavonoids Positive ----
3 Saponins Positive ----
4 Protein and amino acids Positive ----
5 Fats and oil ---- Negative
6 Tannins Positive ----
7 Sterol Positive ----
S. No Tested Phytochemical Positive
Appearance
Negative
Appearance
8 Terpenoids Positive ----
9 Glycosides and anthraquinine Positive ----
10 Resin test ---- Negative
11 Carbohydrates Positive ----
12 Phenol Positive ----
The Veronica genus includes tannins, phenol, saponins,
carboxylic acid, steroids, and flavonoids contents due to which it
shows influential action [11]. Phytochemical are essential dynamic
compounds existing in plants example is flavonoids, alkaloids,
tannins, steroids polyphenol, terpenoids, and saponins,
significantly utilized to prescribe diseases additionally employed
in nutrient plus dietary-supplement [46-50]. Every effective
phytochemical are accountable for a biological action such as
against pathogens provide help in unlike compound development
[51-53]. Flavonoids is essentially polyphenol their bearing can
enhance the potential of the antibiotic toward microbes [54, 55].
Particular flavonoids structure complexes with the cell wall of
bacteria-protein and extracellular ingredients and is extremely
valuable plus efficient composite [56]. Some medicinally
bioactive compound appearance inside plant largely shows
antimicrobial activity possibly it bears flavonoids, tannins,
Saponins, steroids, and terpenoids [57]. Terpenoids concerned
meanwhile reducing microorganism cell wall plus membranous
tissue extinction [58]. Steroid in antimicrobial capable of liposome
leakage of lipid bilayer membrane [59].Saponins synergy amidst
microbes produces enzyme protein leakage of the cell [60]. The
confirmation of phytochemical screening in specie veronica biloba
is for first time reported.
3.3. Antioxidant Activity by DPPH.
The antioxidant activity of medicinal plant a specie
veronica biloba is evaluated according to prescribed procedure of
[34-36] to confirm whether the extracted fractions possess
hydrogen plus free radical scavenging potency because in order to
eliminate the oxidative stress and its effects to keep the organisms
[61]. All the extracted fractions of plants showed excellent
antioxidants results in comparison to the standard as shown in
[Table 3] the concentration used for activity (range from 31.25 to
500µg/mL). Primary, the ethyl-acetate extracted fraction showed
highest IC50 = 1.70±0.05µg/mL inhibition potential and is almost
very closed to the standard positive control Propyl gallate showed
IC50 = 1.6±0.05µg/mL percent potency and is near to other
standard as shown in [Figure 2]. Secondary, dichloromethane
extract fraction showed IC50 = 2.13±0.05µg/mL percent inhibition
which is also near to standard used. While other extract fractions
water showed IC50 = 6.8±0.15µg/mL and similarly n-hexane
showed IC50 = 5.57±0.1µg/mL percent inhibition and standard
ascorbic acid IC50 = 1.27±0.01µg/mL also of 3-Tert-butyl-4-
hydroxyanisol IC50 = 1.2±0.1µg/mL inhibition as shown as shown
in [Table 3]. Our current results aresimilar to [31, 62] when
The antioxidant activity and phytochemical analysis of medicinal plant Veronica biloba
Page | 735
compared we have concluded that this species possess an
antioxidant potential and is the first report on it. An unusual
balance among antioxidant maintenance and the generation of
reactive oxygen species. The oxidative extension may additionally
result in both the raised generation of reactive oxygen species or
from lack of antioxidants [alpha tocopherol, ascorbate or
glutathione]. Inside our body Free radicals mean generated if our
body cells are flashed to numerous substances like chemicals,
radiation, fumes, sun and pesticide whiskey, smoke contamination,
and also due to several metabolic processes including a deposited
fat loss to obtain energy. Irregular intake continues likewise
accountable toward the creation of free radicals. Free radicals
react with protein plus lipids and likewise with
glycosaminoglycans an extracellular matrix [For example Iduronic
acid] and produce associated diseases. Polyunsaturated fatty acids
also amino acids include sulfur are predominantly sensitive. While
the succeeding is observed in large concentrations inside the
CNS, moreover, a study on the use of free radicals in cerebral
ischemia has been conveyed on these molecules. Free radicals
frequently charge at the alpha-methylene carbons that are close to
the carbon-carbon double bond inside fatty acids. Especially
polyunsaturated fatty acids having certain dual bonds in molecules
are consequently capable of free radical impairment [63].
Figure 2. Comparison of Inhibitory Concentration (IC50 Values) Each
Extracted fraction.
Table 3. Antioxidant activity results of Veronica Biloba fraction extracts.
S. NO Fraction Extracts IC50± SD (µg/mL)
1 n-hexane Fraction 5.57 ± 0.1
2 Water Fraction 6.8 ± 0.15
3 Dichloromethane Fraction 2.13 ± 0.05
4 Ethyl-acetate Fraction 1.70 ± 0.05
5 Propyl Gallate(Positive control) 1.6 ± 0.05
6 3-Tert-butyl-4-hydroxyanisol (Positive control) 1.2 ± 0.1
7 Ascorbic acid (Positive control) 1.27 ± 0.01
The data calculated using test statistics as Mean ± SD (a standard deviation). Median inhibitory concentration
(IC50) taken percent inhibition versus different concentration via standard software programmer.
4. CONCLUSIONS
Our current study has shown that a medicinal plant
veronica biloba fraction extracts contain all the important types of
phytochemicals and exhibit the antioxidants inhibition potential.
All the extracts obtained with a good yield mainly in polar parts
and primitive phenolic compound flavonoid is present in this
plant, shows potency towards antioxidants. Our ethyl acetate
extract fraction showed highest inhibition potential at IC50 =
1.70±0.05µg/mL which is much closer to a standard positive
control Propyl gallate showed IC50 = 1.6±0.05µg/mL percent
potential. The purification, isolation and characterization of these
extract areimportant because natural antioxidants currently in
cosmetics products, food and therapeutics health related products
significantly demanded because they are very effective, efficient
and harmless.
5. REFERENCES
1. Taskova, R.M.;Kokubun,T.; K. G. Ryan, P. J. Garnock-
Jones, and S. R. Jensen,Phenylethanoid and iridoid glycosides in
the New Zealand snow hebes (Veronica,
Plantaginaceae).Chemical and Pharmaceutical Bulletin 2010,
585, 703-711,
2. Davis, P. Flora of Turkey and the East Aegean Islands.
University Press, Edinburgh 1978, 6, 682-683.
3. Harput, U.S.;Genç,Y.; Khan, N.; Saracoglu, I. Radical
scavenging effects of different Veronica species.Records of
Natural Products 2011, 52, 100.
4. Graham, J.; Quinn, M.; Fabricant, D.; Farnsworth, N. Plants
used against cancer–an extension of the work of Jonathan
Hartwell.Journal of ethnopharmacology 2000, 73, 347-377,
https://doi.org/10.1016/S0378-8741(00)00341-X.
5. Kibwage, I.O.;Mwangi, J.; Thoithi, G. Quality control of
herbal medicines, East and Central African Journal of
Pharmaceutical Sciences 2005, 82, 27-30,
6. Suffredini, I.; Sader, H.S.; Gonçalves, A.G.; Reis, A.O.;
Gales, A.C.; Varella, A.D.; Younes, R.N. Screening of
antibacterial extracts from plants native to the Brazilian Amazon
Rain Forest and Atlantic Forest. Brazilian journal of medical
and biological research 2004, 373, 379-384,
http://dx.doi.org/10.1590/S0100-879X2004000300015.
7. Cos, P.; Vlietinck, A.J.; Berghe, D.V.; Maes, L. Anti-
infective potential of natural products: how to develop a stronger
in vitro ‘proof-of-concept’. Journal of ethnopharmacology 2006,
106, 290-302, https://doi.org/10.1016/j.jep.2006.04.003.
8. Soldati, F. The registration of medicinal plant products, what
quality of documentation should be required? The industrial
point of view." pp. 1997.
9. Rahman, K.S.S.F.S.A.;Omar, D. Antibacterial activity of the
crude extract of Piper sarmentosum against Pseudomonas
fuscovaginae. International Journal of Applied Biology and
Pharmaceutical Technology 2016,7, 67–72.
10. Bakhtiar, M.S.M.S.I.;Akhter, R.; Bhuiyan, M.A. In vitro
antioxidant activities of the whole plant extract of Chrozophora
prostrata (dalz.).Annals of Biological Research 2015, 6.
11. Gusev, N.;Nemereshina,O.; Petrova,G.; Sychev,M.
Evaluation of antibacterial activity and biologically active
substances of herbal drugs from Veronica L. Russian Journal of
Biopharmaceuticals 2012, 4, 17-22.
Amir Hassan, Himayat Ullah, Muhammad Israr
Page | 736
12. Mocan, A.; Vodnar, D.; Vlase, L.; Crișan, O.; Gheldiu, A.M.;
Crișan, G. Phytochemical characterization of Veronica
officinalis L., V. teucrium L. and V. orchidea Crantz from
Romania and their antioxidant and antimicrobial properties.
International journal of molecular sciences 2015, 16, 21109-
21127, https://doi.org/10.3390/ijms160921109.
13. Dunkic, V.; Kosalec, I.; Kosir, I.J.; Potocnik, T.; Cerenak,
A.; Koncic, M.Z.; Vitali, D.; Muller, I.D.; Kopricanec, M.;
Bezic, N. Antioxidant and antimicrobial properties of Veronica
spicata L.(Plantaginaceae).Current drug targets 2015, 16, 1660-
1670, https://doi.org/10.2174/1389450116666150531161820.
14. Exarchou, V.; Kanetis, L.; Charalambous, Z.; Apers, S.;
Pieters, L.; Gekas, V.; Goulas, V. HPLC-SPE-NMR
characterization of major metabolites in Salvia fruticosa Mill.
Extract with antifungal potential: Relevance of carnosic acid,
carnosol, and hispidulin. Journal of agricultural and food
chemistry 2015, 63, 457-463, https://doi.org/10.1021/jf5050734.
15. Sharifi-Rad, J.; Iriti, M.; Setzer, W.N.; Sharifi-Rad, M.;
Roointan, A.; Salehi, B. Antiviral activity of Veronica persica
Poir. on herpes virus infection. Cellular and Molecular Biology
2018, 64, 11-17, https://doi.org/10.14715/cmb%2F2018.64.8.2.
16. Teng, J.; Li, H.Q.; Yao, Z.; Zhang, Y.W.; Zhamg, F.G.;
Duan, H.Q. Anticancer activity of diterpenes from Veronica
sibirica in vitro.Chinese Traditional and Herbal Drugs 2008, 7.
17. Moreno-Escobar, J.A.; Alvarez, L.; Rodríguez-López, V.;
Bahena, S.M. Cytotoxic glucosydic iridoids from Veronica
americana.Phytochemistry letters 2013, 6, 610-613,
https://doi.org/10.1016/j.phytol.2013.07.017.
18. Živković, J.; Barreira, J.C.; Stojković, D.; Ćebović, T.;
Santos-Buelga, C.; Maksimović, Z.; Ferreira, I.C. Phenolic
profile, antibacterial, antimutagenic and antitumour evaluation
of Veronica urticifolia Jacq.Journal of functional foods 2014, 9,
192-201, https://doi.org/10.1016/j.jff.2014.04.024.
19. Saracoglu, I.; Oztunca, F.H.; Nagatsu, A.; Harput, U.S.
Iridoid content and biological activities of Veronica cuneifolia
subsp. cuneifolia and V. cymbalaria. Pharmaceutical biology
2011, 49, 1150-1157,
https://doi.org/10.3109/13880209.2011.575790.
20. Scarlat, M.; Şandor, V.; Tǎmaş, M.; Cuparencu, B.
Experimental anti-ulcer activity of Veronica officinalis L.
extracts.Journal of ethnopharmacology 1985, 13, 157-163,
https://doi.org/10.1016/0378-8741(85)90003-0.
21. Jeon, H. Anti-inflammatory activity of Veronica
peregrina.Natural Product Sciences 2012, 18, 141-146.
22. Ertas, A.; Boga, M.; Kizil, M.; Ceken, B.; Goren, A.C.;
Hasimi, N.; Demirci, S.; Topcu, G.; Kolak, U. Chemical profile
and biological activities of Veronica thymoides subsp.
Pseudocinerea. Pharmaceutical biology 2015, 53, 334-339,
https://doi.org/10.3109/13880209.2014.919326.
23. Görür, G.; Abdullah, M.; Işik, M. Insecticidal activity of the
Thymus, Veronica and Agrimonia’s essential oils against the
cabbage aphid, Brevicoryne brassicae. Acta Phytopathologica et
Entomologica Hungarica 2008, 43, 201-208,
https://doi.org/10.1556/APhyt.43.2008.1.19.
24. Yin, L.; Lu, Q.; Tan, S.; Ding, L.; Guo, Y.; Chen, F.; Tang,
L. Bioactivity-guided isolation of antioxidant and anti-
hepatocarcinoma constituents from Veronica ciliata. Chemistry
Central Journal 2016, 10, 27,
https://dx.doi.org/10.1186%2Fs13065-016-0172-1.
25. Kwak, J.H.; Kim, H.J.; Lee, K.H.; Kang, S.C.; Zee, O.P.
Antioxidative iridoid glycosides and phenolic compounds from
Veronica peregrina.Archives of pharmacal research 2009, 32,
207-213, https://doi.org/10.1007/s12272-009-1137-x.
26. Ignjatović, D.; Živković, J.; Tovilović, G.; Šavikin, K.;
Tomić, M.; Maksimović, Z.; Janković, T. Evaluation of
angiogenic and neuroprotective potential of different extracts
from three Veronica species.Frontiers in Life Science 2015, 8,
107-116, https://doi.org/10.1080/21553769.2014.998297.
27. Sharifi Rad, M.; Tayeboon, G.; Rad, J.S.; Iriti, M.; Varoni,
E.; Razazi, S. Inhibitory activity on type 2 diabetes and
hypertension key-enzymes, and antioxidant capacity of Veronica
persica phenolic-rich extracts.Cell Mol Biol Noisy-le-grand
2016, 30, 80-5.
28. Stojković, D.S.;Živković, J.; Soković, M.; Glamočlija, J.;
Ferreira, I.C.;Janković, T.; Maksimović, Z. Antibacterial activity
of Veronica montana L. extract and of protocatechuic acid
incorporated in a food system.Food and Chemical Toxicology
2013, 55, 209-213, https://doi.org/10.1016/j.fct.2013.01.005.
29. Sharifi-Rad, J.; Tayeboon, G.S.; Niknam, F.; Sharifi-Rad,
M.; Mohajeri, M.; Salehi, B.; Iriti, M.; Sharifi-Rad, M. Veronica
persica Poir. extract–antibacterial, antifungal and scolicidal
activities, and inhibitory potential on acetylcholinesterase,
tyrosinase, lipoxygenase and xanthine oxidase. Cellular and
Molecular Biology 2018, 64, 50-56,
https://doi.org/10.14715/cmb/2018.64.8.8.
30. Manurung, R.A.H.; Nugroho, R.A.; Sari, Y.P.; Chernovita,
R.; Auliana. Phytochemical Analysis And Antioxidant Activity
Of Leaves Extracts Of Endemic Plant Jahe Balikpapan
(Etlingera Balikpapanensis A.D. Poulsen). International journal
of scientific & technology research 2019, 8
31. Labiad, M.; Harhar, H.; Ghanimi, A.; Tabyaoui, M.
Phytochemical screening and antioxidant activity of Moroccan
thymus satureioides extracts, Journal of Materials and
Environmental Sciences 2017, 8, 2132-2139.
32. Alabri, T.H.A.;Al Musalami, A.H.S.;Hossain, M.A.;Weli,
A.M.;Al-Riyami, Q. Comparative study of phytochemical
screening, antioxidant and antimicrobial capacities of fresh and
dry leaves crude plant extracts of Datura metel L. Journal of
King Saud University-Science 2014, 26, 237-243,
https://doi.org/10.1016/j.jksus.2013.07.002.
33. Oonsivilai, R.; Ferruzzi, M.G.; Ningsanond, S. Antioxidant
activity and cytotoxicity of Rang Chuet (Thunbergia laurifolia
Lindl.) extracts.As J Food Ag-Ind 2008, 102, 116-128.
34. Hossain, M.A.;Shah, M.D.;Gnanaraj, C.; Iqbal, M. In vitro
total phenolics, flavonoids contents and antioxidant activity of
essential oil, various organic extracts from the leaves of tropical
medicinal plant Tetrastigma from Sabah.Asian Pacific journal of
tropical medicine 2011, 4, 717-721,
https://doi.org/10.1016/S1995-7645(11)60180-6.
35. Jacinto,S.D.; Ramos, E.F.;Siguan, A.P.T.;Canoy, R.J.C.
Determining the antioxidant property of plant extracts: a
laboratory exercise. Asian J Biol Edu 2011, 5, 22-5.
36. Marinova, G.; Batchvarov, V. Evaluation of the methods for
determination of the free radical scavenging activity by
DPPH.Bulgarian Journal of Agricultural Science 2011, 17, 11-
24.
37. Khan, A.; Jan, G.; Khan, A.; Jan, F.G.; Bahadur, A.; Danish,
M. In vitro antioxidant and antimicrobial activities of Ephedra
gerardiana (root and stem) crude extract and fractions.Evidence-
Based Complementary and Alternative Medicine 2017, 2017,
https://doi.org/10.1155/2017/4040254. 38. Aiboudi, M.; Yousfi, F.; Fekkar, G.; Bouyazza, L.;
Ramdani, M.; El Azzouzi, M.; Abdel-Rahman, I. Eco-friendly
Allium cepa L. seeds extracts as corrosion inhibitor for mild
steel in 1 M HCl solutions. 2019.
39. Esmaeili, A.K.; Taha, R.M.; Mohajer, S.; Banisalam, B.
Antioxidant activity and total phenolic and flavonoid content of
various solvent extracts from in vivo and in vitro grown
Trifolium pratense L.(Red Clover). BioMed Research
International 2015, 2015.
40. Barchan, A.; Bakkali, M.; Arakrak, A.; Pagán, R.; Laglaoui,
A. The effects of solvents polarity on the phenolic contents and
The antioxidant activity and phytochemical analysis of medicinal plant Veronica biloba
Page | 737
antioxidant activity of three Mentha species extracts.Int J Curr
Microbiol App Sci 2014, 3, 399-412,
41. Latique, S.; Chernane, H.; Mansori, M.; El Kaoua, M.
Seaweed liquid fertilizer effect on physiological and biochemical
parameters of bean plant (Phaesolus vulgaris variety Paulista)
under hydroponic system.European Scientific Journal 2013,
930.
42. Anwar, F.; Przybylski, R. Effect of solvents extraction on
total phenolics and antioxidant activity of extracts from flaxseed
(Linum usitatissimum L.). ACTA Scientiarum Polonorum
Technologia Alimentaria 2012, 11, 293-302.
43. Savithramma, N.; Rao, M.L.; Suhrulatha, D. Screening of
medicinal plants for secondary metabolites.Middle-East Journal
of Scientific Research 2011, 8, 579-584.
44. Chatoui, K.; Talbaoui, A.; Aneb, M.; Bakri, Y.; Harhar, H.;
Tabyaoui, M. Phytochemical Screening, Antioxidant and
Antibacterial activity of Lepidium sativum seeds from
Morocco.J. Mater. Environ. Sci 2016, 78, 2938-2946.
45. El Hattabi, L.; Talbaoui, A.; Amzazi, S.; Bakri, Y.; Harhar,
H.; Costa, J.; Tabyaoui, M. Chemical composition and
antibacterial activity of three essential oils from south of
Morocco (Thymus satureoides, Thymus vulgaris and
Chamaelum nobilis). Journal of Materials and Environmental
Science 2016, 79, 3110-3117.
46. Gordon, D.T.; Stoops, D.; Ratliff, V. Dietary fiber and
mineral nutrition. Eagan Press, Eagan, Minn, USA, 1995.
47. Sanjoaquin, M.A.; Appleby, P.N.; Spencer, E.A.; Key, T.J.
Nutrition and lifestyle in relation to bowel movement frequency:
a cross-sectional study of 20 630 men and women in EPIC–
Oxford.Public Health Nutrition 2004, 7, 77-83,
https://doi.org/10.1079/phn2003522.
48. Topping, D.L.; Clifton, P.M. Short-chain fatty acids and
human colonic function: roles of resistant starch and nonstarch
polysaccharides. Physiological reviews 2001, 813, 1031-1064,
https://doi.org/10.1152/physrev.2001.81.3.1031.
49. Lu, T. Potential health benefits and problems associated with
antinutrients in foods. Food Res. Intern 1993, 26, 131-148,
https://doi.org/10.1016/0963-9969(93)90069-U.
50. P’ei, C.; Chen, S. Verbenaceae.Flora Reipublicae Popularis
Sinicae 1982, 651, 1-49.
51. Sharifi-Rad, J. Herbal Antibiotics: Moving back into the
mainstream as an alternative for" Superbugs". Cellular and
Molecular Biology 2016, 62, 1-2.
52. Alghazeer, R.; El-Saltani, H.; Saleh, N.; Al-Najjar, A.;
Hebail, F. Antioxidant and antimicrobial properties of five
medicinal Libyan plants extracts. Natural science 2012, 4, 324,
https://doi.org/10.4236/ns.2012.45045.
53. Edziri, H.; Mastouri, M.; Cheraif, I.; Aouni, M. Chemical
composition and antibacterial, antifungal and antioxidant
activities of the flower oil of Retama raetam (Forssk.) Webb
from Tunisia.Natural product research 2010, 24, 789-796,
https://doi.org/10.1080/14786410802529190.
54. Sato, Y.; Shibata, H.; Arai, T.; Yamamoto, A.; Okimura, Y.;
Arakaki, N.; Higuti, T. Variation in synergistic activity by
flavone and its related compounds on the increased susceptibility
of various strains of methicillin-resistant Staphylococcus aureus
to β-lactam antibiotics.International journal of antimicrobial
agents 2004, 24, 226-233,
https://doi.org/10.1016/j.ijantimicag.2004.02.028.
55. Cushnie, T.T.; Lamb, A.J. Antimicrobial activity of
flavonoids. International journal of antimicrobial agents 2005,
26, 343-356, https://doi.org/10.1016/j.ijantimicag.2005.09.002.
56. Cowan, M.M. Plant products as antimicrobial agents.Clinical
microbiology reviews 1999, 12, 564-582.
57. Mamtha, B.; Kavitha, K.; Srinivasan, K.; Shivananda, P. An
in vitro study of the effect of Centella asiatica [Indian
pennywort] on enteric pathogens. Indian Journal of
Pharmacology 2004, 36, 41.
58. Hernández, N.E.; Tereschuk, M.; Abdala, L. Antimicrobial
activity of flavonoids in medicinal plants from Tafı del Valle
(Tucuman, Argentina).Journal of Ethnopharmacology 2000, 73,
317-322, https://doi.org/10.1016/s0378-8741(00)00295-6.
59. Epand, R.F.; Savage, P.B.; Epand, R.M. Bacterial lipid
composition and the antimicrobial efficacy of cationic steroid
compounds (Ce ragenins). Biochimica et Biophysica Acta
(BBA)-Biomembranes 2007, 1768, 2500-2509,
https://doi.org/10.1016/j.bbamem.2007.05.023.
60. Zablotowicz, R.M.; Hoagland, R.E.; Wagner, S.C. Effect of
saponins on the growth and activity of rhizosphere bacteria.
Saponins Used in Food and Agriculture 1996, 405, 83-
95,https://doi.org/10.1007/978-1-4613-0413-5_8.
61. Fernández-Agulló, A.; Pereira, E.; Freire, M.S.; Valentao, P.;
Andrade, P.; González-Álvarez, J.; Pereira, P. Influence of
solvent on the antioxidant and antimicrobial properties of walnut
(Juglans regia L.) green husk extracts.Industrial crops and
products 2013, 42, 126-132,
https://doi.org/10.1016/j.indcrop.2012.05.021.
62. Roby, M.H.H.;Sarhan,M.A.; Selim, K.A.H.; Khalel, K.I.
Evaluation of antioxidant activity, total phenols and phenolic
compounds in thyme (Thymus vulgaris L.), sage (Salvia
officinalis L.), and marjoram (Origanum majorana L.)
extracts.Industrial Crops and Products 2013, 43, 827-
831,https://doi.org/10.1016/j.indcrop.2012.08.029.
63. Schmidley, J. Free radicals in central nervous system
ischemia. Stroke 1990, 217, 1086-1090,
https://doi.org/10.1161/01.str.21.7.1086.
64. Hertog, M.G.; Feskens, E.J.; Kromhout, D.; Hertog, M.;
Hollman, P.; Hertog, M.; Katan, M. Dietary antioxidant
flavonoids and risk of coronary heart disease: the Zutphen
Elderly Study. The Lancet 1993, 342, 1007-1011,
https://doi.org/10.1016/0140-6736(93)92876-U.
65. Holiman, P.C.; Hertog, M.G.; Katan, M.B. Analysis and
health effects of flavonoids. Food Chemistry 1996, 57, 43-46,
https://doi.org/10.1016/0308-8146(96)00065-9.
66. Moure, A.; Cruz, J.M.; Franco, D.; omı ng e , J.M.;
Sineiro, J.; omı ng e , H.; José Núñez, M.A.; Parajó, J.C.
Natural antioxidants from residual sources. Food Chemistry
2001, 72, 145-171, https://doi.org/10.1016/S0308-
8146(00)00223-5.
67. Pathak, M.A.; Joshi, P.C. The nature and molecular basis of
cutaneous photosensitivity reactions to psoralens and coal
tar.Journal of Investigative Dermatology 1983, 80, 66s-74s.
68. Halliwell, B. The role of oxygen radicals in human disease,
with particular reference to the vascular system.Pathophysiology
of Haemostasis & Thrombosis 1993, 23, 118-126,
https://doi.org/10.1159/000216921.
69. Meyer, A.S.; Heinonen, M.; Frankel, E.N. Antioxidant
interactions of catechin, cyanidin, caffeic acid, quercetin, and
ellagic acid on human LDL oxidation.Food Chemistry 1998, 61,
71-75, https://doi.org/10.1016/S0308-8146(97)00100-3.
70. Hunt,E.J.; Lester, C.E.; Lester, E.A.; Tackett, R.L. Effect of
St. John's wort on free radical production.Life sciences 2001, 69,
181-190, https://doi.org/10.1016/S0024-3205(01)01102-X.
Amir Hassan, Himayat Ullah, Muhammad Israr
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6. ACKNOWLEDGEMENTS
Amir Hassan thanks to his supervisor Prof. Himayat Ullah of Organic Chemistry Department, Govt. Post Graduate College
Mardan, for his great support and work. Our research work not receives any specific funding from anywhere or any source in the
designing of Study interpretation and data collection analysis, writing and Publication of Manuscript.
© 2019 by the authors. This article is an open access article distributed under the terms and conditions of the
Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).