See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/274250226

Monitoring seasonal variation of epicatechin and gallic acid in the bark of Saraca
asoca using reverse phase high performance liquid chromatography (RP‑HPLC)
method

Article  in  Journal of Ayurveda and integrative medicine · March 2015

DOI: 10.4103/0975-9476.146568

CITATIONS READS

8 135

4 authors, including:

Shraddha umesh Nayak Sandeep Ramchandra Pai

Padmashree Dr.D.Y.Patil Vidyapeeth's Rayat Shikshan Sanstha's Dada Patil Mahavidyalaya
11 PUBLICATIONS   28 CITATIONS    85 PUBLICATIONS   579 CITATIONS   

SEE PROFILE SEE PROFILE

Dr. R. K. Joshi
ICMR-National Institute of Traditional Medicine (Formerly Regional Medical Research…
125 PUBLICATIONS   912 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Plant Diversity and Bioressources of Tunisia View project

Elicitation of triterpenoids from Achyranthes aspera View project

All content following this page was uploaded by Dr. R. K. Joshi on 31 March 2015.

The user has requested enhancement of the downloaded file.

[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

ORIGINAL RESEARCH ARTICLE EXPERIMENTAL

Monitoring seasonal variation of epicatechin

and gallic acid in the bark of Saraca asoca
using reverse phase high performance liquid
chromatography (RP‑HPLC) method
Pushkar M. Ketkar, Shraddha U. Nayak, Sandeep R. Pai1, Rajesh K. Joshi2
Department of Dravyaguna, Karnatak Lingayat Education University, Shri. B. M. Kankanwadi Ayurveda Mahavidyalaya, Belgaum, 1Plant
Biotechnology and Tissue Culture Division, 2Department of Phytochemistry, Regional Medical Research Centre, Indian Council of Medical
Research, Belgaum, Karnataka, India

ABSTRACT
Background: Saraca asoca (Roxb.) Wilde (Fabaceae) is a high valued but vulnerable medicinal plant of Western Ghats
region. This plant is mainly known for its use in various gynecological disorders. Objective: The objective of the present
study was to investigate seasonal variation of the polyphenolic compounds viz., epicatechin and gallic acid in the bark
of S. asoca by using Reverse Phase High Performance Liquid Chromatography‑Diode Array Detector (RP‑HPLC‑DAD)
method. Materials and Methods: The bark was collected in six different Ritu (season) viz. Varsha (monsoon),
Sharad (autumn), hemant (early winter), Shishir (winter), Vasanta (spring), and Grishma (summer) mentioned in Ayurveda.
Results: The RP‑HPLC‑DAD analysis indicated that levels of epicatechin and gallic acid in the bark of S. asoca vary
seasonally. The highest concentration of epicatechin was observed in Shishir Ritu (3315.19 ± 165.76 mg/100g) and
gallic acid during Hemant Ritu (211.90 ± 10.60 mg/100 g). Conclusions: In present study, the ability to synthesize and
accumulate both the compounds in bark of S. asoca varied greatly throughout the seasons. It was also observed that the
compound epicatechin was present abundantly as compared to gallic acid throughout the seasons.

Key words: Ayurveda, epicatechin, gallic acid, Saraca asoca (Roxb.) Wilde, seasonal variation

INTRODUCTION revealed that aqueous and alcoholic extracts of the stem

bark of S. asoca have been reported phenolic glycoside P2
Saraca asoca (Roxb.) Wilde (Fabaceae) commonly known from phenolic glycoside fraction and also non‑phenolic
as ‘Ashoka’ is a highly valued medicinal plant categorized glycosides which have stimulant action on isolated human
‘vulnerable’ by International Union for Conservation of uterus.[3,4] Moreover, the phenolic glycoside P2 was inactive
Nature [Figure 1]. This plant is mainly known in Ayurveda on central nervous system (CNS), cardiovascular system, and
for its use in gynecological disorders.[1,2] Recent studies smooth muscles other than uterus.[3] Furthermore, diverse
Address for correspondence: pharmacological activities viz., antibacterial,[5] anticancer,[6]
Dr. Rajesh K. Joshi, Department of Phytochemistry, Regional antimutagenic, genoprotective effect,[7] antihyperglycemic,[8]
Medical Research Centre, Indian Council of Medical Research, antioxidant, [8,9] molluscicidal, [10] anxiolytic, [11] CNS
Nehru Nagar, Belgaum ‑ 590 010, Karnataka, India.
E‑mail: joshirk_natprod@yahoo.com depressant, [12] anti‑pyretic, [13] and analgesic [14] have
Received: 07‑Mar‑2014 been reported from different parts of S. asoca. This
Revised: 09‑Apr‑2014 plant is used to treat skin infections, CNS function,
Accepted: 17‑May‑2014
genitor‑urinary functions, uterus pain during periods, clots,
and ammenorhea.[15] Polyphenolic compounds catechin,
Access this article online
epicatechin, leucocyanidin, [16] leucopelargonidin, [17]
Quick Response Code: Website: procyanidin B‑2, 11′‑deoxyprocyanidin B,[18] b‑sitosterol,[19]
www.jaim.in
gallic acid, quercetin, (‑) 3‑O‑p‑D‑glucoside,[20] catechol, (‑)
epicatechol[16‑19] apigenin‑7‑O‑b‑D‑glucoside, cyanidin‑3,
DOI: 5‑diglucoside, kaempferol 3‑O‑b‑D‑glucoside,
10.4103/0975-9476.146568
pelarg onidin‑3,5‑diglucoside, quercetin and its
3‑O‑b‑D‑glucoside, n‑octacosanol,  (−)‑procyanidin

Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1 29

[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

Ketkar, et al.: Seasonal variation of epicatechin and gallic acid in the bark of S. asoca

derivatives, methyl‑ and ethylcholesterol derivatives, liquid chromatography  (HPLC) grade acetonitrile,

kaempferol‑3‑O‑a‑L‑rhamnoside, amyrin, and ceryl glacial acetic acid and water were purchased from Fisher
alcohol[19,21] have been identified from the plant of S. asoca. Scientific (Thermo electron LLS India Pvt. Ltd.)
Out of the identified compounds, epicatechin and gallic
acid are the major polyphenolic bioactive molecules in Plant collection and processing
the bark of S. asoca, which have various pharmacological The plant of S. asoca was identified and authenticated at Regional
effects. Gallic acid shows evidence of suppression of Medical Research Centre (RMRC), Belgaum (Voucher No.
a high‑fat diet‑induced dyslipidemia, hepatosteatosis, RMRC 980). The bark was collected from a single habitat
oxidative stress in rats,[22] and also possess many potential from Belgaum region (N 15.862 E 074.510; elevation:
therapeutic properties including anti‑cancer, antimicrobial 799M above MSL) during specified Ritu (season) as given
properties,[23] and neuroprotective action.[24] Epicatechin in Table 1[33,34] from the same trees (n = 3). Flowering trees
protects endothelial cells against oxidized low‑density over 3‑meters height were considered as mature plants for
lipoprotein (LDL) by scavenging free radicals, maintaining collection of bark samples (each tree one bark sample). The
nitric oxide synthase,[25] and possess insulin‑like activity.[26] collected bark (from 3 trees) was shade dried, powdered,
Gallic acid and epicatechin have cholesterol‑lowering activity sieved, and stored in cool and dry place until further use.
by inhibiting pancreatic cholesterol esterase, binding of bile
acids, and reducing solubility of cholesterol in micelles.[27] Extraction of plant material
Extraction was achieved using cold maceration technique for
Ayurveda is the ancient science of medicine. It is suggested all samples. Two‑gram bark powder was accurately weighed
that part specific and Ritu (season) specific collection of plant and soaked in 20 mL methanol overnight. The mixture was
capitulate gives maximum efficacy and potency (Veeryavan).[28] sonicated (Sonicater bath, Bandelin‑Sonorex, Germany,
Ayurveda mentions six Ritus (seasons) viz. Shishira (winter), 35 KHz) for 30 minutes and filtered using Whatman No. 1
Vasanta  (spring), Grishma (summer), Varsha (monsoon), filter paper. The extraction procedure was repeated three
Sharada (autumn), and Hemanta (early winter).[29] The officinal times and pooled. The solvent was distilled off using
part, stem bark (Twak) of S. asoca is used for therapeutic Rotaevaporator at 40ºC. The extracts were stored in sealed
purpose.[30] In Charaka samhita and Sushruta samhita, it has vial at 4ºC until analysis. The concentration of 0.5 mg/
been suggested that the stem bark of medicinal plants should mL was prepared in methanol and was used for RP-HPLC
be collected in Sharada Ritu (Ashwin‑Kartik).[31,32] The aim of analysis after filtering through 0.2 µ Nylon filter paper.
the present study was to analyse the seasonal variations of
the epicatechin and gallic acid in the bark of S. asoca. Table 1: Seasons as per Indian and English
calendars
Year of Indian calendar English calendar
MATERIALS AND METHODS collection Month Ritu Month Season
2012 Shraavana (Sawan) and Varsha July to Monsoon
Chemicals Bhadrapada (Bhado) September
Epicatechin  [(−)‑cis‑3, 3′,4′,5,7-Pentahydroxyflavane] 2012 Ashwin (Kwar) and Sharad September Autumn
Kartika to November
[Figure 2a] and gallic acid [3,4,5‑trihydroxybenzoic
2012‑13 Margashirsha Hemant November to Early
acid] [Figure 2b] were purchased from Natural (Agrahayana, Agahan) January winter
Remedies, Bangalore, India. The high performance and Pausha (Poos)
2013 Magh and Shishir
January to Winter
Phalguna (Phagun) March
2013 Chaitra and Baisakh Vasanta
March to Spring
May
2013 Jyeshta and Aashaadha Grishma May to July Summer

2+ 2 2+

+2 2
2+

2+ +2 2+

2+ 2+
a b
Figure 1: Inflorescence of Saraca asoca Figure 2: Chemical structure of (a) Epicatechin; (b) Gallic acid

30 Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1
[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

Ketkar, et al.: Seasonal variation of epicatechin and gallic acid in the bark of S. asoca

Reverse phase high performance liquid chromatography- acid was recorded [Figure 3 a and b]. A 5‑point standard
diode array detector (RP‑HPLC‑DAD) analysis computer generated linear calibration curve of standard
The RP‑HPLC analysis was performed on Shimadzu epicatechin and gallic acid within the concentration
chromatographic system (Model no. LC‑20AD) consisting of ranges of 0.5‑10 µg/mL and 0.1‑10 µg/mL, respectively
a quaternary pump, manual injector, degasser (DGU‑20A5), was obtained with coefficient of determination (R2)
and dual λ ultraviolet (UV) absorbance diode array not more than 0.982 [Figure 3 c‑d]. The regression
detector (Model No. SPD‑M20A). The built in LC (Liquid equations showed significant relationship between peak
Chromatography) solution software system was used area and concentration and this equation was used to
for data processing. Chromatographic separation was estimate contents from samples. LOD of epicatechin
achieved on a Qualisil BDS (Base Deactivated Silica) and gallic acid were 0.192, 0.490 µg/mL, and LOQ were
250‑4.6 mm (5 µm) C18 column. A mobile phase consisting 0.582, 1.485 µg/mL, respectively. The relative standard
of “A” (acetonitrile), “B” (water), and “C” (glacial acetic deviation (RSD) values were less than 2% indicating
acid) were used for separation with 12:85:3 in an isocratic method to be precise and reproducible. Validation of
mode with injection volume of 20 µL. The flow rate was method was done by spiking 50 µL (5µg/mL) of standards
0.7 mL/min and the detection wavelength of diode array to equal volume of extracts to obtain recovery within the
detector (DAD) was set 280 nm with 18‑minutes run time range of 95‑100%.
for both standard and sample.
The profile of epicatechin and gallic acid using
Calibration and linearity RP‑HPLC‑DAD analysis of the extracts of bark of
Calibration and linearity was achieved by accurately weighing S. asoca was obtained during six different seasons, achieved
epicatechin and gallic acid dissolved it in methanol to obtain as the final output of this study. The results of RP‑HPLC
mg/mL standard stock solution. The stocks were serially quantitative analysis of the content yields of epicatechin
diluted to prepare working solutions (epicatechin: 0.5 to and gallic acid are presented in Figure 4. The analysis
10  µg/mL; gallic acid: 0.1 to 10 µg/mL) for calibration revealed that sample collected during Shishira Ritu contained
curves at five concentration levels separately. All solutions higher level of epicatechin (3315.19 ± 165.76 mg/100g),
were stored at 4ºC temperature. The calibration curve for whereas gallic acid was found in high amount during
the standards with above analytical column was established Hemanta Ritu (211.90 ± 10.60 mg/100g). On the other
by peak areas and concentrations of working solutions. hand, lowest content was recorded during Grishma
Ritu (409.40 ± 20.47 mg/100g) for epicatechin and Vasanta
System suitability Ritu  (30.96 ± 1.55 mg/100g) for gallic acid. More than
The system suitability test was assessed by three 80% difference between highest and lowest content in
replicate injections of the standard solutions at a certain epicatechin and gallic acid were observed [Figure 4].
concentration. The peak areas of epicatechin and gallic acid
were used to evaluate repeatability of the method and their
DISCUSSION
peaks were analyzed for resolution and tailing factors. The
limits of detection (LOD) and quantification (LOQ) were As per Ayurveda, potency of the bark drug corresponding
determined with the signal/noise method. Signal/noise to the action should be at its peak during Sharada Ritu.
ratios of 3.3 and 10 were applied for estimating the LOD Thus it is expected that the contents responsible for this
and LOQ, respectively. action must be abundant in the respective season. But in
the present investigation Shishira and Hemanta Ritu were
RESULTS responsible for yielding higher content of epicatechin and
gallic acid, respectively. The quantitative and qualitative
The yields of extracts at different seasons were divergence may be due to the climatic conditions, which
34.88 ± 1.74% (Shishira); 20.32 ± 1.02% (Vasanta); in turn may affect the composition and other secondary
9.33 ± 0.47 (Grishma); 22.34 ± 1.12% (Varsha); metabolites of the plants.[35,36] Phenolic compounds are said
19.86 ± 0.99% (Sharada); 33.00 ± 1.65% (Hemanta). The to be maximum during the summer season which is also
identification of the two polyphenols viz., epicatechin evident from the present study.[37] The compounds may
and gallic acid in the extract of S. asoca, was done by accumulate at a particular period in the plants with response
comparing retention times and UV spectral data with to environmental changes.[37,38] Biological activity which
those of authentic standards. The profiles of both is dependent on the chemical composition, is similarly
samples and standards are presented in Figure 3 (a‑e). subject to variation depending on the said factors.[39]
Profiles with retention time of 16.056 ± 0.158 minutes Ayurveda emphasizes on standardization of the crude drug
for epicatechin and 6.208 ± 0.040 minutes for gallic as well as the end product. The time of collection, place

Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1 31

[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

Ketkar, et al.: Seasonal variation of epicatechin and gallic acid in the bark of S. asoca

P$8 P$8
QPQP QPQP
 








(SLFDWHFKLQXJPO




 

*DOOLFDFLGXJPO
 









 

 

 



        PLQ                    PLQ

a b
 



 
 \ [  \ [
 5  5  


 
             
Epicatechin µg/ml *DOOLFDFLG—JPO
c d

e
Figure 3: HPLC chromatograms of (a) Standard epicatechin 5 µg/mL; (b) gallic acid 5 µg/mL; (c-d) Five-point calibration curves of the standard
epicatechin and gallic acid; (e) HPLC profile of samples collected during different Ritu or season. HPLC: High performance liquid chromatography




(3,

 *$


PJJ



























9DUVKD 6KDUDG +HPDQW 6KLVKLU 9DVDQWD *ULVKPD

Figure 4: Comparative histogram of epicatechin and gallic acid in the bark of S. asoca. EPI: Eicatechin, GA: Gallic acid

32 Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1
[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

Ketkar, et al.: Seasonal variation of epicatechin and gallic acid in the bark of S. asoca

of collection, methodology is documented in the ancient 8. Kumar S, Narwal S, Kumar D, Singh G, Narwa S, Arya R.
Evaluation of antihyperglycemic and antioxidant activities
texts. All these factors are basically being told for utilizing of Saraca asoca (Roxb.) De Wild leaves in streptozotocin
the maximum potency of the medicine.[40] Apart from the induced diabetic mice. Asian Pac J Trop Dis 2012;2:170‑6.
phytochemical group of substances typical for a taxon, 9. Saha J, Mukherjee S, Gupta K, Gupta B. High‑performance
the chemical outfit depends on the specific genotype, the thin‑layer chromatographic analysis of antioxidants present
in different parts of Saraca asoca (Roxb.) de Wilde. J Pharm
stage of plant development, influence of environmental Res 2013;7:798‑803.
factors and the part of the plant.[41] The variation in the 10. Singh A, Singh VK. Molluscicidal activity of Saraca asoca
secondary metabolites among plants chemotype may occur and Thuja orientalis against the fresh water snail Lymnaea
acuminata. Vet Parasitol 2009;164:206‑10.
within different sites. This may also be due to genetic 11. Manohar VR, Mohandas R, Chandrashekar R. Acute anxiolytic
responses to climatic and edaphic factors in formation of effect of ethanolic extract of Saraca asoka bark in wistar
secondary metabolites.[42,43] albino rats. Int J Bioassays 2013;2:926‑8.
12. Verma A, Jana GK, Sen S, Chakraborty R, Sachan S,
Mishra A. Pharmacological evaluation of Saraca indica leaves
CONCLUSION for central nervous system depressant activity in mice.
J Pharm Sci Res 2010;2:338‑43.
13. Varaprasad N, Suresh A, Suresh V, Kumar SN, Rajendar A,
It was observed that Ritu or season influences secondary Madeshwaran M, et al. Anti pyretic activity of methanolic
metabolite concentration in the plants or its parts. In extract of Saraca asoca (Roxb.) de Wild leaves. Int J Pharm
Res Dev 2011;3:202‑7.
present study, the ability to synthesize and accumulate 14. Verma A, Jana GK, Chakraborty R, Sen S, Sachan S,
both the compounds in bark of S. asoca varied greatly Mishra A. Analgesic activity of various leaf extracts of Saraca
throughout the seasons. Apart from this, it was also indica Linn. Pharm Lett 2010;2:352‑7.
observed that the compound epicatechin was present 15. Nadkarni KM. The Indian Materia Medica. 3rd ed. Bombay:
Popular Book Depot; 1957. p. 1075.
abundantly as compared to gallic acid throughout the 16. Indrani N, Balasubramanian K. Isolation of condensed tannins
seasons. Consequently, to obtain optimum utility of the from Saraca ashoka. Part I. Leather Sci 1984;31:349‑51.
plant as a source of medicine, the screening for presence of 17. Kaur DJ, Misra K. Leucoanthocynidins from Saraca ashoka
stem bark. J Indian Chem Soc 1980;57:1243.
the secondary metabolites during different season is useful. 18. Middelkoop TB, Labadie RP, Proanthocynidins in the bark of
Further studies are required to determine and correlate the Saraca asoca Roxb. De Wilde. Z Naturforsch 1985;40B: 855‑7.
biological activities with respect to the concentration of 19. Behari M, Andhiwal CK, Ballantine JA. Phytosterols from the
bark of Saraca indica Linn. Indian J Chem 1977;15B: 765‑6.
compounds present in the plant/plant parts. 20. Sadhu SK, Khatun A, Phattanawasin P, Ohtsuki T,
Ishibashi  M. Lignan glycosides and flavonoids from Saraca
asoca with antioxidant activity. J Nat Med 2007;61:480‑2.
ACKNOWLEDGMENTS 21. Khare CP. Indian Medicinal Plants. An Illustrated Dictionary.
Berlin/Heidelberg: Springer Science, Springer‑Verlag; 2007.
Authors are thankful to the Dr. B. S. Prasad, Principal, KLE’U 22. Hsu CL, Yen GC. Effect of gallic acid on high fat diet‑induced
Shri. B. M. Kankanwadi Ayurveda Mahavidyalaya, Belgaum and dyslipidaemia, hepatosteatosis and oxidative stress in rats. Br
J Nutr 2007;98:727‑35.
Officer‑in‑Charge, RMRC, Belgaum for providing necessary
23. Ow YY, Stupans I. Gallic acid and gallic acid derivatives:
facilities. Effects on drug metabolizing enzymes. Curr Drug
Metab 2003;4:241‑8.
24. Bastianetto S, Yao ZX, Papadopoulos V, Quirion R.
REFERENCES Neuroprotective effects of green and black teas and their
catechin gallate esters against beta‑amyloid‑induced toxicity.
1. Billore KV, Yelne MB, Dennis TJ, Chaudhari BG. Data Base Eur J Neurosci 2006;23:55‑64.
on Medicinal plants used in Ayurveda. Vol. 4. New Delhi: 25. Steffen Y, Schewe T, Sies H. Epicatechin protects endothelial
CCRAS; 2005. p. 423‑32. cells against oxidized LDL and maintains NO synthase.
2. Chakrapanidatta Chakradatta. In: Sharma PV, editor. 3rd ed. Biochem Biophys Res Commun 2005;331:1277‑83.
Varanasi: Chaukhamba Publishers; 2002. p. 524. 26. Rizvi SI, Zaid MA. Insulin‑like effect of (‑) epicatechin on erythrocyte
3. Satyavati GV, Prasad DN, Sen SP, Das PK. Oxytocic activity membrane acetylcholinesterase activity in type 2 diabetes
of a pure phenolic glycoside (P2) from Saraca indica Linn mellitus. Clin Exp Pharmacol Physiol 2001;28:776‑8.
(Ashoka): A short communication. Indian J Med Res 27. Ngamukote S, Mäkynen K, Thilawech T, Adisakwattana S.
1970;58:660‑3. Cholesterol‑lowering activity of the major polyphenols in
4. Satyavati GV, Prasad DN, Sen SP, Das PK. Further studies on grape seed. Molecules 2011;16:5054‑61.
the uterine activity of Saraca indica Linn. Indian J Med Res 28. Acharya YT. Charaka Samhita of Agnivesha; Kalpasthana;
1970;58:947‑60. Madanakalpa. Reprint ed. ch. 1. Ver. 7. Varanasi:
5. Pal SC, Maiti AP, Chatterjee BP, Nandy A. Antibacterial Chaukhamba Orientalia; 2011. p. 652.
activity of flowers and flower buds of Saraca indica Linn. 29. Sharma R, Amin H, R G, Prajapati PK. Seasonal variations in
Indian J Med Res 1985;82:188‑9. physicochemical profiles of Guduchi Satva (starchy substance
6. Varghese CD, Satish, Nair C, Panikkar KR. Potential anticancer from Tinospora cordifolia [Willd.] Miers). J Ayurveda Integr
activity of Saraca asoca extracts towards transplantable Med 2013;4:193‑7.
tumours in mice. Indian J Pharm Sci 1992;54:37‑40. 30. Sharma PC, Yelne MB, Dennis TJ. Database on medicinal plants
7. Nag D, Ghosh M, Mukherjee A. Antimutagenic and used in Ayurveda, Vol. 3. New Delhi: CCRAS; 2001. p. 76.
genoprotective effect of Saraca asoca bark extract. Toxicol 31. Acharya YT. Charaka Samhita of Agnivesha; Kalpasthana;
Ind Health 2013. doi: 10.1177/0748233713483200. Madanakalpa. Reprint ed. Ch. 1 verse 10. Varanasi:

Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1 33

[Downloaded free from http://www.jaim.in on Monday, March 30, 2015, IP: 14.99.4.220]  ||  Click here to download free Android application for this journal

Ketkar, et al.: Seasonal variation of epicatechin and gallic acid in the bark of S. asoca

Chaukhamba Orientalia; 2011. p. 653. Seasonal variation in content, chemical composition and
32. Acharya YT. Sushruta Samhita of Sushruta; Sutrasthana; antimicrobial and cytotoxic activities of essential oils from
Bhumipravibhagiya. Reprint ed. ch. 37. Ver. 5. Varanasi: four Mentha species. J Sci Food Agric 2010;90:1827‑36.
Chaukhamba Sanskrita Sansthana; 2010. p. 159. 40. Acharya YT. Charaka Samhita of Agnivesha; Vimanasthana;
33. Raghavan V. Ritu in Sanskrit literature; Shri Lal Bahadur Rogabhishakjitiya. Reprint ed. Ch. 8. Ver. 144. Varanasi:
Shastri Kendriya Sanskrit Vidyapeetha. Delhi; 1972. Chaukhamba Orientalia; 2011. p. 762.
34. Available from: http://en.wikipedia.org/wiki/Ritu_(Indian_ 41. Franz C. Genetics. In: Hay RK, Waterman PG, editors. Volatile
season). [Last accessed on 2014 May 4]. Oil Crops. Harlow: Longman; 1993. p. 63‑96.
35. Joshi RK, Badakar VM, Kholkute SD, Khatib N. Chemical 42. Hanover JW. Genetics of terpenes. I. Gene control of
composition and antimicrobial activity of the essential oil of monoterpene levels in Pinus monticola Dougl. Heredity
the leaves of Feronia elephantum (Rutaceae) from north west 1966;21:73‑84.
Karnataka. Nat Prod Commun 2011;6:141‑3. 43. Mithen R, Raybould AF, Giamoustaris A. Divergent selection
36. Joshi RK. Pulegone and menthone chemotypes of Mentha for secondary matabolites between wild populations of
spicata Linn. from Western Ghats region of North West Brassica oleracea and its implications for plant‑herbivore
Karnataka, India. Natl Acad Sci Lett 2013;36:349‑52. interactions. Heredity 1995;75:472‑84.
37. Pai SR, Pawar NV, Nimbalkar MS, Kshirsagar PR, Kolar FK,
Dixit GB. Seasonal variation in content of camptothecin
from the bark of Nothapodytes nimmoniana (Grah.)
Mabb., using HPLC analysis. Pharmacognosy Res 2013; How to cite this article: Ketkar PM, Nayak SU, Pai SR, Joshi
5:219‑23. RK. Monitoring seasonal variation of epicatechin and gallic acid in
38. Pai SR, Nimbalkar MS, Pawar NV, Patil RP, Dixit GB. Seasonal the bark of Saraca asoca using reverse phase high performance
discrepancy in phenolic content and antioxidant properties liquid chromatography (RP-HPLC) method. J Ayurveda Integr Med
from bark of Nothapodytes nimmoniana (Grah.) Mabb. Int J 2015;6:29-34.
Pharma Bio Sci 2010;1:3:1‑8. Source of Support: Nil, Conflict of Interest: None declared.
39. Hussain AI, Anwar F, Nigam PS, Ashraf M, Gilani AH.

34 Journal of Ayurveda & Integrative Medicine | January-March 2015 | Vol 6 | Issue 1

View publication stats