Antioxidant potential and impact of different extraction solvents on the free, esterified and insoluble-bound phenolics, flavonoid and tannin content of Trillium govanianum Wall ex D. Don, a rare Himalayan herb

, , ,

Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-022-00365-8
First Page: 953
Last Page: 960
Views: 237

Keywords: Trillium , Bioactive yield, Antioxidant, FRAP, Phenolic content, Flavonoid


Trillium govanianum is a compelling medicinal herb that mainly occurs in the sub-alpine and alpine regions of the Himalayan states of India. It has acquired a threatened status due to large-scale illicit exploitation because of its effective medicinal properties. A limited number of studies on the antioxidant potential of T. govanianum along with its establishment as a source of new antioxidants led us to carry out this investigation. The study also investigated the efficacy of three different solvents on the quantitative extraction of bioactive compounds from the herb for its maximum utilization. For the study, rhizomes of T. govanianum were subjected to enzymatic and non-enzymatic assays. Analysis of enzymes catalase (CAT) and ascorbate peroxidase (AP) was carried out by simple phosphate buffer extraction method, phenolics and tannin determination by Folin–Ciocalteu method, flavonoid content by aluminium chloride colorimetric method and also total reducing and ferric reducing power assays were done. Results showed that rhizomes of T. govanianum exhibit fair CAT and AP activity. From the results it was also apparent that absolute methanol was the most feasible solvent among the all three used, to get the highest solid extractable yield (40.24%) as well as total phenolics and tannins (46.7 ± 0.15 and 0.74 ± 0.01 mg/gm of extract respectively) whereas acetone extract yielded more flavonoid content (16.5 ± 0.15 mg/gm of extract). FRAP activity and total reducing power of the methanol extract was also higher than the other two extracts. These findings reveal a promising potential of this herb as a source of antioxidants comprehending a need for its cultivation and conservation.

              , Bioactive yield, Antioxidant, FRAP, Phenolic content, Flavonoid

*Pdf Download

(*Only SPR Members can download pdf file; #Open Access;)


Acosta-Estrada BA, Gutierrez-Uribe JA, Serna-Saldivar SO (2014) Bound phenolics in foods, a review. Food Chem 152:46–55.

Aebi H (1983) Catalase in vitro. Methods Enzymol 105:121–126

Agbo MO, Uzorl PF, Akazie-Nneji UN, Eze-Odurukwe CU, Ogbatue UB, Mbaoji EC (2015) Antioxidant, total phenolic and flavonoid content of selected Nigerian medicinal plants. Dhaka Univ J Pharm Sci 14(1):35–41

Arruda HS, Pereira GA, de Morais DR, Eberlin MN, Pastore GM (2018) Determination of free, esterified, glycosylated and insoluble-bound phenolics composition in the edible part of araticum fruit ( Annona crassiflora Mart.) and its by-products by HPLC-ESI-MS/MS. Food Chem 245:738–749.

Benzie IFF, Strain JJ (1999) Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol 299:15–27

Chandra S, Singh A, Singh CP, Nautiyal MC, Rawat LS (2018) Vascular plants distribution in relation to topography and environmental variables in the alpine zone of Kedarnath Wild Life Sanctuary, West Himalaya. J Mt Sci 15(9):1936–1949

Chandran KCI, Indira G (2016) Quantitative estimation of total phenolic, flavonoids, tannin and chlorophyll content of leaves of Strobilanthes Kunthiana (Neelakurinji). J Med Plants Stud 4(4):282–286

Chauhan NS (1999) Medicinal and aromatic plants of Himachal Pradesh. Indus Publishing Company, New Delhi

Cheynier V (2012) Phenolic compounds: from plants to foods. Phytochem Rev 11:153–177.

Corpas FJ, Barroso JB, del Río LA (2001) Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells. Trends Plant Sci 6:145–150

Dai J, Mumper RJ (2010) Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules 15(10):7313–7352

D’Archivio M, Filesi C, Di Benedetto R, Gargiulo R, Giovannini C, Masella R (2007) Polyphenols, dietary sources and bioavailability. Ann Ist Super Sanita 43(4):348–361

Das S, Talukdar D, Sangha MK, Chaudhary DP, Borah N, Das A, Das S, Saikia SP (2017) Antioxidant enzymes potential in leaves of oats and barley and phytochemistry of stress tolerance. J Pharmacogn Phytochem 1:694–703

Do Diem Q, Angkawijaya AE, Tran-Nguyen PL, Huynh LH, Soetaredjo FE, Ismadji S, Ju Y-H (2014) Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatic. J Food Drug Anal 22(3):296–302

Ezez D, Tefera M (2021) Effects of solvents on total phenolic content and antioxidant activity of ginger extracts. J Chem.

Ghasemzadeh A, Jaafar Hawa ZE, Rahmat A (2010) Antioxidant activities, total phenolics and flavonoids content in two varieties of Malaysia young ginger (Zingiber officinale Roscoe). Molecules 15:4324–4333

Haminiuk CWI, Plata-Oviedo MSV, de Mattos G, Carpes ST, Branco IG (2014) Extraction and quantification of phenolic acids and flavonols from Eugenia pyriformis using different solvents. J Food Sci Technol 51(10):2862–2866

Kenganora M, Bhaskaran M, Santhepete MN, Kumar V, Hukkeri I (2017) Antioxidant potential of a toxic plant Calotropis procera R. Br. Free Radic Antioxid 7(2):143–151

Krygier K, Sosulski F, Hodge L (1982) Free, esterified, and insoluble bound phenolic acids. 1. Extraction and purification procedure. J Agric Food Chem 30:330–334

Liyana-Pathirana CM, Shahidi F (2006) Importance of insoluble-bound phenolics to antioxidant properties of wheat. J Agric Food Chem 54(4):1256–1264.

Mahmood A, Mahmood A, Malik RN (2012) Indigenous knowledge of medicinal plants from Leepa valley, Azad Jammu and Kashmir, Pakistan. J Ethnopharmacol 143(1):338–346

Medini F, Fellah H, Ksouri R, Abdelly C (2014) Total phenolic, flavonoid and tannin contents and antioxidant and antimicrobial activities of organic extracts of shoots of the plant Limonium delicatulum. J Taibah Univ Sci 8:216–224

Mueller-Harvey I (2006) Unraveling the conundrum of tannins in animal nutrition and health. J Sci Food Agric 86:2010–2037

Naczk M, Shahidi F (1989) The effect of methanol-ammonia-water treatment on the content of phenolic acids of canola. Food Chem 31:159–164

Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880

Ngo TV, Scarlett CJ, Bowyer MC, Ngo PD, Vuong QV (2017) Impact of different extraction solvents on bioactive compounds and antioxidant capacity from the root of Salacia chinensis L. J Food Qual 1:1–8

Nguyen NVT, Duong NT, Nguyen KNH, Bui NT, Pham TLT, Nguyen KT, Le PH, Kim KH (2021) Effect of extraction solvent on total phenol, flavonoid content, and antioxidant activity of Avicennia officinalis. Biointerface Res Appl Chem 12(2):2678–2690

Oyaizu M (1986) Studies on product of browning reaction prepared from glucose amine. Jpn J Nutr 44:307–315

Pant S, Samant S (2010) Ethnobotanical observations in the Mornaula reserve forest of Kumaun, West Himalaya, India. Ethnobot Leaflets 14:193–217

Rahman SU, Ismail M, Shah MR, Iriti M, Shahid M (2015) GC/MS analysis, free radical scavenging, anticancer and β-glucuronidase inhibitory activities of Trillium govanianum rhizome. Bangladesh J Pharmacol 10:577–583

Rahman SU, Adhikari A, Ismail M, Shah MR, Khurram M, Shahid M, Ali F, Haseeb A, Akbar F, Iriti M (2016) Beneficial effects of Trillium govanianum rhizomes in pain and inflammation. Molecules 21:1095

Samant SS, Dhar U, Palni LMS (1998) Medicinal plants of indian himalaya: diversity, distribution potential values. Gyanodaya Prakashan, Nainital

Shahidi F, Ju-Dong Y (2016) Insoluble-bound phenolics in food. Molecules 21:1216.

Singleton V, Rossi J (1965) Colorimetry of total phenolic compounds with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

Truong DH, Nguyen DH, Ta Anh NT, Vo Bui A, Ha Do T, Nguyen HC (2019) Evaluation of the use of different solvents for phytochemical constituents, antioxidants, and in vitro anti-inflammatory activities of Severinia buxifolia. J Food Q.

Widyawati PS, Budianta TDW, Kusuma FA, Wijaya EL (2014) Difference of solvent polarity to phytochemical content and antioxidant activity of Pluchea indica less leaves extracts. Int Pharmacol Phytochem Res 6(4):850–855



We gratefully acknowledge our heartfelt gratitude to Head, HAPPRC, colleagues at lab and field staff for their help and support.

Author Information

Chandola Vaishali
High Altitude Plant Physiology Research Center, H. N. B. Garhwal University, Garhwal, India
Chandra Sudeep
High Altitude Plant Physiology Research Center, H. N. B. Garhwal University, Garhwal, India

Nautiyal A. R.
High Altitude Plant Physiology Research Center, H. N. B. Garhwal University, Garhwal, India
Concenço Germani
Brazilian Agricultural Research Corporation, EMBRAPA, Temperate Agriculture, Pelotas, Brazil