Search JIM Advanced Search

Journal of Integrative Medicine ›› 2016, Vol. 14 ›› Issue (6): 447-455.doi: 10.1016/S2095-4964(16)60276-5

• Research Article • Previous Articles     Next Articles

Determination of flavonoids, polyphenols and antioxidant activity of Tephrosia purpurea: A seasonal study

Madan Mohan Pandey, Sayyada Khatoon, Subha Rastogi, Ajay Kumar Singh Rawat   

  1. Pharmacognosy & Ethnopharmacology Division, CSIR-National Botanical Research Institute, Lucknow 226 001, India
  • Received:2016-05-19 Accepted:2016-08-04 Online:2016-11-15 Published:2016-11-15
  • Contact: Subha Rastogi, PhD; E-mail:


Tephrosia purpurea (Linn.) Pers. is widely used in traditional medicine to treat liver disorders, febrile attacks, enlargement and obstruction of liver, spleen, and kidney. In the present study, investigations were carried out to determine the seasonal impact on the content of flavonoid glycosides and on antioxidant activities so as to identify the optimal time of harvesting.


The plant materials were collected in different seasons during 2013-2014. Air-dried, powdered plant materials were extracted with 95% ethanol and ethanol:water (1:1) by ultrasound-assisted extraction process. Their chemical composition in terms of total polyphenol and flavonoid contents (TPCs and TFCs) was determined using modified colorimetric Folin-Ciocalteu method and aluminum chloride colorimetric assay respectively. To determine the in vitro antioxidant activity, diphenyl-picryl hydrazyl (DPPH) radical-scavenging assay and total antioxidant capacity by phosphomolybdate antioxidant assay were carried out. High-performance liquid chromatography (HPLC)/photo-diode array (PDA) analysis was used to quantify the flavonoid glycosides in the samples collected in different seasons. Correlation studies were also carried out between antioxidant activities and TPCs.


The highest TPC and TFC were found to be in the 95% ethanolic extract of the August sample and the lowest in the 50% hydro-alcoholic extract of the plant sample collected in winter season. It was observed that in both the assays used to determine the antioxidant activity, the 95% ethanolic extracts in all the seasons showed a higher activity than their respective 50% hydro-alcoholic extracts with an increase in activity as we go from cold to hot to rainy seasons. Based on correlation analysis, DPPH radical-scavenging activities as well as the spectrophotometrically measured phosphomolybdenum complex were also strongly correlated with TPC of the extracts. The most abundant flavonoid glycoside was quercetin-3-O-rhamnoglucoside in all the seasons. The content of all flavonoids was observed highest in the 95% ethanolic extract of the plant collected in August (TP-3). The 50% hydro-alcoholic extract of the plant collected in December (TP-6) showed the lowest amount of flavonoids and antioxidant activity.


The findings of this study confirmed that the metabolism and production of flavonoids in T. purpurea are vigorously affected by seasonal factors. Significant differences were observed in the TPC, TFC and flavonoid glycoside composition in the 95% ethanolic as well as 50% hydro-alcoholic extracts of T. purpurea collected in different seasons. Since the 95% ethanolic extract of the August sample (TP-3), which also happens to be its flowering season, exhibited the highest TPC, TFC and antioxidant activities in both DPPH and phosphomolybdate assays as well as contained the highest content of all flavonoids, it could be recommended as the optimal season of harvesting T. purpurea with respect to its pharmaceutically important constituents, i.e., flavonoids.

Key words: Tephrosia, Chromatography, High-performance liquid, Flavonoid glycosides, Seasonal variation, Antioxidant effect

1 Chopra RN, Nayar SL, Chopra IC.Glossary of Indian medicinal plants. New Delhi: Council of Scientific and Industrial Research. 1956.
2 Sankaran JR.Tefroli in the management of viral hepatitis.Antiseptic. 1980; 77: 643-646.
3 Kumar A, Dutta M, Bhatt TK, Dalal DS.Use of herbal tonic Yakrifi t in equine practice.Indian Vet J. 1997; 74(5): 424-425.
4 Kirtikar KR, Basu BD.Indian medicinal plants. 2nd ed. Allahabad: LM Basu. 1935: 724. 5 Despande SS, Shah GB, Parmar NS. Antiulcer activity of Tephrosia purpurea in rats. Indian J Pharmacol. 2003; 35: 168-172.
6 Anonymous. The wealth of India—a dictionary of Indian raw materials and industrial product. New Delhi: Council of Scientifi c and Industrial Research. 1976: 151-156.
7 Zafar R, Mujeeb M, Ahmed S.Preliminary phytochemical screening of root culture ofTephrosia purpurea(Linn.) Pers. Hamdard Med. 2004; 48: 1.
8 Gokhale AB, Saraf MN.Tephrosia purpurea: a review of contemporary literature and medicinal properties. Indian Drugs. 2000; 37: 553-560.
9 Lodhi S, Pawar RS, Jain AP, Singhai AK.Wound healing potential ofTephrosia purpurea(Linn.) Pers. in rats. J Ethnopharmacol. 2006; 108(2): 204-210.
10 Sharma P, Rastogi S, Bhatnagar S, Srivastava JK, Dube A, Guru PY, Kulshrestha DK, Mehrotra BN, Dhawan BN.Antileishmanial action ofTephrosia purpurea Linn. extract and its fractions against experimental visceral leishmaniasis. Drug Dev Res. 2003; 60: 285-293.
11 Pelter A, Ward RS, Rao EV, Raju NR.8-Substituted fl avonoids and 3'-substituted 7-oxygenated chalcones fromTephrosia purpurea. J Chem Soc. 1981; 9: 2491-2498.
12 Kumar S, Pandey AK.Chemistry and biological activities of flavonoids: an overview.Sci World J. 2013; 2013: 162750.
13 Badarinath AV, Rao KM, Chetty CMS, Ramkanth S, Rajan TVS, Gnanaprakash K.A review onin vitro antioxidant methods: comparisions, correlations and considerations. Int J Pharm Tech Res. 2010; 2(2): 1276-1285.
14 Pal D, Verma P.Flavonoids: a powerful and abundant source of antioxidants.Int J Pharm Pharm Sci. 2013; 5(3): 95-98.
15 Liu LS, Crowe NL, Ju HY, Pong DF, Liu WS, Zhang JY.Variation of fl avonoids and other phenolics in the leaves of Chinese plum (Prunus salicina Lindl.) cultivars. Advances Hortic. 1994; 472-480.
16 Akabori Y.Flavonoid pattern in the pteridaceae IV. Seasonal variation of the flavonoids in the fronds ofAdiantum monochlamys. J Plant Res. 1978; 91(2): 137-139.
17 Siegelman HW.Physiological studies on phenolic compounds. In: Harborne JB. Biochemistry of phenolic compounds. New York: Academic Press. 1964: 437-456.
18 Harborne JB.Comparative biochemistry of the fl avonoids. New York: Academic Press. 1967.
19 Graham TL.Flavonoid and isoflavonoid distribution in developing soybean seedling tissues and in seed and root exudates.Plant Physiol. 1991; 95(2): 594-603.
20 Chaves N, Escurdero JC, Gutierrez-Merino C.Seasonal variation of exudate ofCistus ladanifer. J Chem Ecol. 1993; 19(11): 2577-2591.
21 Stefkov G, Karapandzova M, Stefova M, Kulevanova S.Seasonal variation of flavonoids inTeucrium polium L.(Lamiaceae). Maced Pharm Bull. 2009; 55(1 -2): 33-40.
22 Modak B, Torres R, Urzúa A.Seasonal variation of the flavonoids pinocembrin and 3-O-methylgalangin, in the surface component mixture (resinous exudates and waxy coating) ofHeliotropium stenophyllum. J Chil Chem Soc. 2011; 56(1): 532-534.
23 Costa GM, Ortmann CF, Schenkel EP, Reginatto FH.Seasonal variations in the amount of isoorientin and isovitexin inCecropia glaziovii Sneth. leaves over a twoyear period. Rev Colomb Cienc Quím Farm. 2014; 43(1): 162-172.
24 Wolfe K, Wu X, Liu RH.Antioxidant activity of apple peels.J Agric Food Chem. 2003; 51(3): 609-614.
25 Ordo ñz AAL, Gomez JD, Vattuone MA, Isla MI.Antioxidant activities ofSechium edule(Jacq.) Swart extracts. Food Chem. 2006; 97(3): 452-458.
26 Sreejayan N, Rao MN.Free radical scavenging activity of curcuminoids.Arzneimittel-forschung. 1996; 46(2): 169-171.
27 Prieto P, Pineda M, Aguilar M.Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E.Anal Biochem. 1999; 269(2): 337-341.
28 Prior RL, Wu X, Schaich K.Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements.J Agric Food Chem. 2005; 53(10): 4290-4302.
29 Chua LS.A review on plant-based rutin extraction methods and its pharmacological activities.J Ethnopharmacol. 2013; 150(3): 805-817.
30 Leong CN, Tako M, Hanashiro I, Tamaki H.Antioxidant flavonoid glycosides from the leaves ofFicus pumila L. Food Chem. 2008; 109(2): 415-420.
31 Yang JX, Guo J, Yuan JF.In vitro antioxidant properties of rutin. LWT-Food Sci Technol. 2008; 41(6): 1060-1066.
32 Agrawal AD.Pharmacological activities of flavonoids: a review.Int J Pharm Sci Nanotech. 2011; 4(2): 1394-1398.
33 Damre AS, Gokhale AB, Phadke AS, Kulkarni KR, Saraf MN.Studies on the immunomodulatory activity of flavonoidal fraction ofTephrosia purpurea. Fitoterapia. 2003; 74(3): 257-261.
34 Tapas AR, Sakarkar DM, Kakde RB.Flavonoids as nutraceuticals: a review.Trop J Pharm Res. 2008; 7(3): 1089-1099.
[1] Abdulhakim Abubakar, Abdullahi Balarabe Nazifi, Idris Mohammed Maje, Yusuf Tanko, Joseph Akpojo Anuka, Ezzeldin Mukthar Abdurahman. Antihyperglycaemic activity of ethylacetate extract of Chlorophytum alismifolium in type 2 diabetes: The involvement of peroxisome proliferator-activated receptor-γ and dipeptidyl peptidase-4. Journal of Integrative Medicine, 2021, 19(1): 78-84.
[2] Rabie Kachkoul, Tarik Squalli Housseini, Mohamed Mohim, RadouaneEl Habbani, Youssef Miyah, Anissa Lahrichi. Chemical compounds as well as antioxidant and litholytic activities of Arbutus unedo L. leaves against calcium oxalate stones. Journal of Integrative Medicine, 2019, 17(6): 430-437.
[3] Sonali S. Nipate, Pramila S. Yelmar. Therapeutic potential of ethyl acetate fraction of Tephrosia purpurea Linn. leaves in a rat model of gout. Journal of Integrative Medicine, 2019, 17(6): 455-460.
[4] Asmae Assaidi, Ikram Dib, Monique Tits, Luc Angenot, Said Bellahcen, Nourelhouda Bouanani, Abdelkhaleq Legssyer, Mohammed Aziz, Hassane Mekhfi, Mohammed Bnouham, Michel Frederich, Abderrahim Ziyyat. Chenopodium ambrosioides induces an endothelium-dependent relaxation of rat isolated aorta. Journal of Integrative Medicine, 2019, 17(2): 115-124.
[5] Rohitash Jamwal. Bioavailable curcumin formulations: A review of pharmacokinetic studies in healthy volunteers. Journal of Integrative Medicine, 2018, 16(6): 367-374.
[6] Shohini Chakraborty, Nashra Afaq, Neelam Singh, Sukanta Majumdar. Antimicrobial activity of Cannabis sativaThuja orientalis and Psidium guajava leaf extracts against methicillin-resistant Staphylococcus aureus. Journal of Integrative Medicine, 2018, 16(5): 350-357.
[7] Sandeep Kumar Singh, Arjun Patra. Evaluation of phenolic composition, antioxidant, anti-inflammatory and anticancer activities of Polygonatum verticillatum (L.). Journal of Integrative Medicine, 2018, 16(4): 273-282.
[8] Nanaware Sadhana, Sathiyanarayanan Lohidasan, Kakasaheb Ramoo Mahadik. Marker-based standardization and investigation of nutraceutical potential of Indian propolis. Journal of Integrative Medicine, 2017, 15(6): 483-494.
[9] Bhanuz Dechayont, Arunporn Itharat, Pathompong Phuaklee, Jitpisute Chunthorng-Orn, Thana Juckmeta, Nuntika Prommee, Nitra Nuengchamnong, Pintusorn Hansakul. Antioxidant activities and phytochemical constituents of Antidesma thwaitesianum Müll. Arg. leaf extracts. Journal of Integrative Medicine, 2017, 15(4): 310-319.
[10] Yan-bin Shi, Hui-li Li, Hai-qin Wang, Yan-biao Yang, Xiao-yun Zhang, Hui Wang, Zong-jie Zhu, Zhi-ye Zhang, Cheng-an Zhang. Simultaneous determination of five anthraquinones in a Chinese traditional preparation by RP-HPLC using an improved extraction procedure. Journal of Integrative Medicine, 2014, 12(5): 455-462.
[11] Fu-feng Li, Jie Zhao, Peng Qian, Yi-qin Wang, Jing-jing Fu, Zhu-mei Sun, Hai-xia Yan, Li Yang. Metabolite changes in the greasy tongue coating of patients with chronic gastritis. Journal of Chinese Integrative Medicine, 2012, 10(7): 757-765.
[12] Hui-qing Zhang , Zi-fei Yin , Jia-yu Sheng , Zhu-qian Jiang , Bao-yu Wu , Yong-hua Su. A comparison study of pharmacokinetics between bufalin-loaded bovine serum albumin nanoparticles and bufalin in rats. Journal of Chinese Integrative Medicine, 2012, 10(6): 674-680.
[13] Kumaradoss Mohan Maruga Raja Muthu, Hari Mishra Shri. Isolation, characterization and thin-layer chromatography method development of clerosterol palmityl ester: a chemical marker for standardization of leaves of Clerodendrum phlomidis. Journal of Chinese Integrative Medicine, 2012, 10(1): 109-113.
[14] Venkatapura C. Devaraj, Burdipad G. Krishna, Gollapalle L. Viswanatha. Simultaneous determination of quercetin, rutin and kaempferol in the leaf extracts of Moringa oleifera Lam. and Raphinus sativus Linn. by liquid chromatography-tandem mass spectrometry. Journal of Chinese Integrative Medicine, 2011, 9(9): 1022-1030.
[15] Guo-di Lu, Jing Zhou, Hong-yue Ma, De-kang Wu, Da-wei Qian, Yu-ping Tang, Ming Hong, Yong-qing Hua, Jing-ao Duan. Analysis on competitive interaction between arenobufagin and verapamil hydrochloride with serum albumin. Journal of Chinese Integrative Medicine, 2011, 9(7): 768-774.
Full text



[1] Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2] Zhao-guo Li. Discussion on English translation of commonly used sentences in traditional Chinese medicine: part one. Journal of Chinese Integrative Medicine, 2008, 6(1): 107-110
[3] Jun Hu, Jian-ping Liu. Non-invasive physical treatments for chronic/recurrent headache. Journal of Chinese Integrative Medicine, 2008, 6(1): 31
[4] Xue-mei Liu, Qi-fu Huang, Yun-ling Zhang, Jin-li Lou, Hong-sheng Liu, Hong Zheng. Effects of Tribulus terrestris L. saponion on apoptosis of cortical neurons induced by hypoxia-reoxygenation in rats. Journal of Chinese Integrative Medicine, 2008, 6(1): 45-50
[5] . Uniform requirements for manuscripts submitted to biomedical journals: Writing and editing for biomedical publication (Chinese version, part two). Journal of Chinese Integrative Medicine, 2010, 8(11): 1001-1005
[6] Daniel Weber, Janelle M Wheat, Geoffrey M Currie. Inflammation and cancer: Tumor initiation, progression and metastasis,and Chinese botanical medicines. Journal of Chinese Integrative Medicine, 2010, 8(11): 1006-1013
[7] Hong Liu , Guo-liang Zhang, Li Shen , Zhen Zeng, Bao-luo Zhou, Cheng-hai Liu, Guang Nie . Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5Nl avian influenza virus. Journal of Chinese Integrative Medicine, 2010, 8(11): 1036-1040
[8] Zhao-guo Li . A discussion of English translation of 1995 and 1997 Chinese National Standards of Traditional Chinese Medical Terminologies for Clinical Diagnosis and Treatment. Journal of Chinese Integrative Medicine, 2010, 8(11): 1090-1096
[9] Rui Jin, Bing Zhang. A complexity analysis of Chinese herbal property theory: the multiple formations of herbal property (Part 1). Journal of Chinese Integrative Medicine, 2012, 10(11): 1198-1205
[10] Hui-min Liu, Xian-bo Wang, Yu-juan Chang, Li-li Gu. Systematic review and meta-analysis of randomized controlled trials of integrative medicine therapy for treatment of chronic severe hepatitis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1211-1228