Assessment of optimal infusion condition for Thunbergia laurifolia tea by using response surface methodology

Piyanuch - Rojsanga, Puriwat - Jiewatakuntum, Waree - Limwikrant, Kotchaphan - Chooluck


According to the National List of Essential Medicines of Thailand, the Thunbergia laurifolia (TL) tea is categorized as antipyretic and detoxifying herbal medicine. This tea has also been used for the treatment of drug addiction and smoking cessation. However, suggested infusion conditions were varied which may result in variations in extracted active contents and bioactivities. In this study, the infusion condition that maximized extraction of caffeic acid (CA) and rosmarinic acid (RA), the major markers of TL tea, was assessed by using response surface methodology (RSM). The contents of markers were analyzed by a validated high-performance liquid chromatographic method. To evaluate the extraction efficiency, the marker contents obtained by preparing the tea under the optimal condition were compared to those obtained by aqueous extraction process. The results demonstrated that the marker contents varied considerably according to infusion variables; time, temperature and volume. By using RSM, it was successfully found that the optimal condition for the tea (3 g) was as follows: 200 mL of boiling water and infusion time of 10 min. More importantly, by using infusion method, the major portion of markers remained in the tea powder implying poor extraction efficiency of the method. This result suggested that the use of TL extract would be benefit in terms of quality and efficacy. The optimal infusion condition could be utilized in clinical studies to find an effective oral dose range of markers. Ultimately, the obtained information could be used as support information for determining the oral dose of TL extract.


Caffeic acid, Infusion condition, Response surface methodology, Rosmarinic acid, Thunbergia laurifolia


National List of Essential Medicines, Available at:, acessed May 2018.

P Tejasen and C Thongthapp. The study of the insecticide antitoxicity of Thunbergia laurifolia Linn. Chiang Mai Med. Bull. 1980; 19,105-14.

S Ussanawarong, T Thesiri, T Mahakunakorn and S Parasupattana. Effect of Thunbergia laurifolia Linn. on detoxication of paraquat. Khon Kaen Univ. Res. J. 2000; 5, 11-7.

S Ussanawarong and T Thesiri. Effect of Thunbergia laurifolia Linn. on detoxication of parathion in rat. Khon Kaen Univ. Res. J. 2001; 6, 3-13.

MP Phyu and J Tangpong. Protective effect of Thunbergia laurifolia (Linn.) on lead induced acetylcholinesterase dysfunction and cognitive impairment in mice. Biomed. Res .Int. 2013; 2013, 186098.

J Tangpong and S Satarug. Alleviation of lead poisoning in the brain with aqueous leaf extract of the Thunbergia laurifolia (Linn.). Toxicol. Lett. 2010; 198, 83-8.

W Ruangyuttikarn, P Chattaviriya, N Morkmek, S Chuncharunee and N Lertprasertsuke. Thunbergia laurifolia leaf extract mitigates cadmium toxicity in rats. Scienceasia. 2013; 39, 19-25.

A Chanawirat, C Toshulkao, P Temcharoen and T Glinsukon. 2000, The protective effect of Thunbergia laurifolia extract on ethanol-induced hepatotoxicity in mice. Master of Science Thesis in Toxicology, Mahidol University, Thailand.

P Pramyothin, H Chirdchupunsare, A Rungsipipat and C Chaichantipyuth. Hepatoprotective activity of Thunbergia laurifolia Linn extract in rats treated with ethanol: in vitro and in vivo studies. J. Ethnopharmacol. 2005; 102, 408-11.

O Wonkchalee, T Boonmars, C Aromdee, P Laummaunwai, W Khunkitti, K Vaeteewoottacharn, P Sriraj, R Aukkanimart, W Loilome, Y Chamgramol, C Pairojkul, Z Wu, A Juasook and P Sudsarn. Anti-inflammatory, antioxidant and hepatoprotective effects of Thunbergia laurifolia Linn. on experimental opisthorchiasis. Parasitol. Res. 2012; 111, 353-9.

P Suwanchaikasem, C Chaichantipyuth and S Sukrong. Antioxidant-guided isolation of rosmarinic acid, a major constituent, from Thunbergia laurifolia and its use as a bioactive principle for standardization. Chiang Mai J. Sci. 2014; 41, 117-27.

N Ruangpayungsak. 2016, Development of HPLC method for fingerprinting and quantitative analysis of active compounds of Thunbergia Laurifolia leaf extract using chemometric analysis. Master of Science Thesis in Pharmacy, Mahidol University, Thailand.

VR Coelho, CG Vieira, LP de Souza, F Moyses, C Basso, DK Papke, TR Pires, IR Siqueira, JN Picada and P Pereira. Antiepileptogenic, antioxidant and genotoxic evaluation of rosmarinic acid and its metabolite caffeic acid in mice. Life Sci. 2015; 122, 65-71.

M Ozarowski, PL Mikolajczak, A Bogacz, A Gryszczynska, M Kujawska, J Jodynis-Liebert, A Piasecka, H Napieczynska, M Szulc, R Kujawski, J Bartkowiak-Wieczorek, J Cichocka, T Bobkiewicz-Kozlowska, B Czerny and PM Mrozikiewicz. Rosmarinus officinalis L. leaf extract improves memory impairment and affects acetylcholinesterase and butyrylcholinesterase activities in rat brain. Fitoterapia. 2013; 91, 261-71.

P Rojsanga, P Sithisarn, K Tanaka, D Mizuki and K Matsumoto. Thunbergia laurifolia extract ameliorates cognitive and emotional deficits in olfactorectomized mice. Pharm. Biol. 2015; 53, 1141-8.

H Takeda, M Tsuji, M Inazu, T Egashira and T Matsumiya. Rosmarinic acid and caffeic acid produce antidepressive-like effect in the forced swimming test in mice. Eur. J. Pharmacol. 2002; 449, 261-7.

H Takeda, M Tsuji, J Miyamoto and T Matsumiya. Rosmarinic acid and caffeic acid reduce the defensive freezing behavior of mice exposed to conditioned fear stress. Psychopharmacology (Berl). 2002; 164, 233-5.

X Jin, P Liu, F Yang, YH Zhang and D Miao. Rosmarinic acid ameliorates depressive-like behaviors in a rat model of CUS and Up-regulates BDNF levels in the hippocampus and hippocampal-derived astrocytes. Neurochem. Res. 2013; 38, 1828-37.

S Kondo, AE Omri, J Han and H Isoda. Antidepressant-like effects of rosmarinic acid through mitogen-activated protein kinase phosphatase-1 and brain-derived neurotrophic factor modulation. J. Funct. Foods. 2015; 14, 758-66.

W Thongsaard and C Marsden. A herbal medicine used in the treatment of addiction mimics the action of amphetamine on in vitro rat striatal dopamine release. Neurosci. Lett. 2002; 329, 129-32.

W Thongsaard, C Marsden, P Morris, M Prior and YB Shah. Effect of Thunbergia laurifolia, a Thai natural product used to treat drug addiction, on cerebral activity detected by functional magnetic resonance imaging in the rat. Psychopharmacology (Berl). 2005; 180, 752-60.

W Thongsaard and C Marsden. Effect of Thunbergia laurifolia extract on extracellular dopamine level in rat nucleus accumbens. J. Med. Assoc. Thai. 2013; 96 Suppl 1, S85-9.

W Thongsaard, R Sangpayap and C Marsden. Chronic effect of Thunbergia laurifolia extract and cocaine in rats using behavior model of addiction. J. Med. Assoc. Thai. 2015; 98 Suppl 9, S48-52.

Rosmarinic acid for smoker dehabituation, Available at:, acessed May 2018.

Agent For Smoker Dehabituation, Available at:, accessed May 2018.

L Barros, M Duenas, MI Dias, MJ Sousa, C Santos-Buelga and IC Ferreira. Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions. Food Chem. 2013; 136, 1-8.

A Atoui, A Mansouri, P Kefalas and G Boskou. Tea and herbal infusions: their antioxidant activity and phenolic profile. Food Chem. 2005; 89, 27-36.

X Su, J Duan, Y Jiang, X Duan and F Chen. Polyphenolic profile and antioxidant activities of oolong tea infusion under various steeping conditions. Int. J. Mol. Sci. 2007; 8, 1196-205.

TFF da Silveira, AD Meinhart, CA Ballus and HT Godoy. The effect of the duration of infusion, temperature, and water volume on the rutin content in the preparation of mate tea beverages: an optimization study. Food Res. Int. 2014; 60, 241–5.

E Sharpe, F Hua, S Schuckers, S Andreescu and R Bradley. Effects of brewing conditions on the antioxidant capacity of twenty-four commercial green tea varieties. Food Chem. 2016; 192, 380-7.

Z Nikniaz, R Mahdavi, SJ Ghaemmaghami, N Lotfi Yagin and L Nikniaz. Effect of different brewing times on antioxidant activity and polyphenol content of loosely packed and bagged black teas (Camellia sinensis L.). Avicenna J. Phytomed. 2016; 6, 313-21.

JT DiPiro. Concepts in Clinical Pharmacokinetics. American Society of Health-System Pharmacists, Bethesda; 2010.

Association of official analytical chemistry (AOAC). Guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. Available at:, accessed May 2018.

MF Nishiyama, MAF Costa, AM Costa, CGM Souza, CG Bôer and CK Bracht. Brazilian green tea (Camellia sinensis var assamica): effect of infusion time, mode of packaging and preparation on the extraction efficiency of bioactive compounds and on the stability of the beverage. Ciênc. Tecnol. Aliment. 2010; 30, 191-6.

JE Cacace and G Mazza. Mass transfer process during extraction of phenolic compounds from milled berries. J. Food. Eng. 2003; 59, 379-89.

JR Vergara-Salinas, J Perez-Jimenez, JL Torres, E Agosin, JR Perez-Correa. Effects of temperature and time on polyphenolic content and antioxidant activity in the pressurized hot water extraction of deodorized thyme (Thymus vulgaris). J. Agric. Food Chem. 2012; 60, 10920-9.

YS Lin, YJ Tsai, JS Tsay and JK Lin. Factors affecting the levels of tea polyphenols and caffeine in tea leaves. J. Agric. Food Chem. 2003; 51, 1864-73.

CI Heck and EG de Mejia. Yerba mate tea (Ilex paraguariensis): a comprehensive review on chemistry, health implications, and technological considerations. J. Food Sci. 2007; 72, 138-51.

S Govindaraghavan, NJ Sucher. Quality assessment of medicinal herbs and their extracts: criteria and prerequisites for consistent safety and efficacy of herbal medicines. Epilepsy Behav. 2015; 52, 363-71.


  • There are currently no refbacks.

Online ISSN: 2228-835X

Last updated: 20 June 2019