Optimization formulation of chamomile-based functional beverage as hypnotic agent using response surface method

Annisa Ulfah Pristya, Tri Dewanti Widyaningsih


Insomnia problem can be treated by consuming herbs containing apigenin and EGCG. Chamomile and green tea water extract have been found to containapigenin and EGCG. Optimization of the formulation of those herbs is necessary to obtain the highest hypnotic activity. The purpose of this study was to optimize formulation of chamomile, green tea, and cinnamon to obtain functional beverage containing the highest phenolic and flavonoid content, but the lowest caffeine content. The best formula of the herbs was then tested in caffeine-induced mice to see the hypnotic effect. The research design in this experiment was Response Surface Model (RSM) using Central Composite Design (CCD) method. The in vivo test used Post Test Only Control Group Design. Consisting five groups of 30 male mice divided to 6 mice for each group. The optimum formula for functional beverages suggested by RSM was chamomile: green tea : cinnamon of 70.32 : 30.35 : 4.99. The verification results showed that the actual response of this optimum formula had a total phenol of 23.76 mgGAE/g dry herbs, total flavonoids of 126.43 mgQE/g dry herbs, caffeine content of 29.87 mg caffeine/g dry herbs, apigenin levels of 0.07±0.068 µg/g dry herbs, and EGCG of 6.43±0.218 µg/g dry herbs. Functional beverages showed hypnotic activity in group of mice dosed 26 mL/kg bw, which significantly reduced motor activity to 4.83±0.72 sec. As well as significantly reducing the proinflammatory cytokines 


Apigenin; EGCG; Functional beverages; Insomnia

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Adachi, N., Tomonaga, S., Tachibana, T., Denbow, D.M.,Furuse,M., (2006) '(−)-Epigallocatechin gallate attenuates acute stress responses through GABAergic system in the brain', Eur. J. Pharmacol, 531, pp. 171–175

Akinseye, O.R., Yetunde, A., (2016) 'The Influence of Camiella sinesis, Matricaria chamomilla and their Blend Teas Infusion on Serum Lipids of Wistar Rats', Int. J. Pharmacol. Phytochem. Ethnomedicine, 4, pp. 7–13

An, L., Li, J., Yu, S.-T., Xue, R., Yu, N.-J., Chen, H.-X., Zhang, L.-M., Zhao, N., Li, Y.-F., Zhang, Y.-Z., (2015), 'Effects of the total flavonoid extract of Xiaobuxin-Tang on depression-like behavior induced by lipopolysaccharide and proinflammatory cytokine levels in mice', J. Ethnopharmacol. 163, pp. 83–87

Anggraini, D.T., Prihanta, W., Purwanti, E., (2015) 'Penggunaan Ekstrak Batang Kayu Manis (Cinnamomum burmannii) Terhadap Kualitas Minuman Nata de Coco 7', Seminar Nasional XII Pendidikan Biologi FKIP UNS 2015. [In Indonesian]

Anggriawan, M.B., Roswiem, A.P., Nurcholis, W., n.d. 'The Potency of Aqueous and Ethanolic Bark Extracts of Cinnamon Padang (Cinnamomum burmanii) Againts a-Glukosidase Enzyme Activities 12', Jurnal Kedokteran YARSI 23 (2), pp. 091-102

AOAC: Official Methods of Analysis (Volume 1),1995. 771

Aoyama, K., Nakaki, T., (2013) 'Neuroprotective properties of the excitatory amino acid carrier 1 (EAAC1)', Amino Acids 45, pp. 133–142

Arwangga, A.F., n.d. Analisis Kandungan Kafein Pada Kopi Di Desa Sesaot Narmada Menggunakan Spektrofotometri UV-VIS. Jurnal Kimia (Journal of Chemistry) [In Indonesian]

Ashton, H., (2005) 'The diagnosis and management of benzodiazepine dependence', Curr. Opin. Psychiatry, 18, pp. 249–255

Atanassova, M., Georgieva, S., Ivancheva, K., (2011) 'Total phenolic and total flavonoid contents, antioxidant capacity and biological contaminants in medicinal herbs', Journal of the University of Chemical Technology and Metallurgy, 46(1), pp. 81-88

Baldessarini, R.J., Tondo, L., Hennen, J., (2006) 'Treating the Suicidal Patient with Bipolar Disorder: Reducing Suicide Risk with Lithium', Ann. N. Y. Acad. Sci, 932, 24–43

Banks, S., (2007), 'Behavioral and physiological consequences of sleep restriction', J. Clin. Sleep Med. 3(5), pp. 519–528

Bansode, P.A., (2015) 'Total flavonoid content of commonly consumed teas in india', World J. Pharm. Res, 4, 874–881

Bhattacharya, S., Dey, P., Chandra, S., Chatterjee, P., (2011) 'Neuropharmacological properties of Mikania scandens (L.) Willd. (Asteraceae)', J. Adv. Pharm. Technol. Res. 2, 255

Bhosale, U., Pophale, P., Somani, R., Yegnanarayan, R., Zambare, M., (2011) 'Study of central nervous system depressant and behavioral activity of an ethanol extract of Achyranthes aspera (Agadha) in different animal models' Int. J. Appl. Basic Med. Res. 1, pp. 104

Binti Ismail, N.N., Sapina Abdullah, U.U.@, (2015) Determination of phenolic and antioxidant properties in tea and spent tea under various extraction method and determination of catechins, caffeine and gallic acid by HPLC. Int. J. Adv. Sci. Eng. Inf. Technol. 5, pp. 158.

Burgos, I., Richter, L., Klein, T., Fiebich, B., Feige, B., Lieb, K., Voderholzer, U., Riemann, D., (2006) 'Increased nocturnal interleukin-6 excretion in patients with primary insomnia: A pilot study', Brain. Behav. Immun. 20, pp.246–253.

Bystritsky, A., Hovav, S., Sherbourne, C., Stein, M.B., Rose, R.D., Campbell-Sills, L., Golinelli, D., Sullivan, G., Craske, M.G., Roy-Byrne, P.P., (2012) 'Use of complementary and alternative medicine in a large sample of anxiety patients', Psychosomatics, 53, pp. 266–272

Cabrera, C., Giménez, R., López, M.C., (2003) 'Determination of tea components with antioxidant activity. J. Agric. Food Chem. 51, pp. 4427–4435.

Campbell, N.A., Reece, J.B., Urry, L.A., Wasserman, S.A., Cain, M.L., Minorsky, P.V., (2004) Biologi, 5th ed, 3, Erlangga, Jakarta. [In Indonesian]

Campêlo, L.M.L., Lima, S.G. de, Feitosa, C.M., Freitas, R.M. de, (201) 'Evaluation of central nervous system effects of citrus limon essential oil in mice', Rev. Bras. Farmacogn. 21, pp. 668–673

Chavez Valdez, R., Ahlawat, R., Wills-Karp, M., Nathan, A., Ezell, T., Gauda, E.B., (2011) 'Correlation between Serum Caffeine Levels and Changes in Cytokine Profile in a Cohort of Preterm Infants', J. Pediatr. 158, 57-64.e1

Cleverdon, R., Elhalaby, Y., McAlpine, M., Gittings, W., Ward, W., (2018) 'Total polyphenol content and antioxidant capacity of tea bags: comparison of black, green, red rooibos, chamomile and peppermint over different steep times', Beverages 4, pp. 15

Connole, L., Harkin, A., Maginn, M., (2004) 'Adenosine A1 receptor blockade mimics caffeine’s attenuation of ethanol-induced motor incoordination. basic htmlent glyphamp asciiamp clin. pharmacol. Htmlent Glyphamp Asciiamp Toxicol. 95, pp. 299–304

Deacon, R.M.J., (2013) 'Measuring motor coordination in mice', J. Vis. Exp.

Department of Biology, Faculty of Mathemathic and Natural Sciences, University of Brawijaya, Malang, Indonesia, Dwijayanti, D.R., Rifa’i, M., (2014) 'Immunomodulator testing on ethanol extract of Gynura procumbens leaves to Mus musculus adaptive immune system: in vitro study', J. Exp. Life Sci. 4, pp. 10–14

Dhawan, K., (2003) 'Attenuation of benzodiazepine dependence in mice by a tri-substituted benzoflavone moiety of Passiflora incarnata Linneaus: A non-habit forming anxiolytic', J Pharm Pharm Sci, 6(2),pp. 215-22

Everson, C.A., Laatsch, C.D., Hogg, N., (2005) 'Antioxidant defense responses to sleep loss and sleep recovery. Am. J. Physiol.-Regul. Integr'. Comp. Physiol, 288, pp. R374–R383

Folin, O., Ciocalteu, V., (1944) 'Tyrosine and tryptophane determinations in proteins. J. Biol. Chem. 73, pp. 627–650

Friedman, M., Kim, S.-Y., Lee, S.-J., Han, G.-P., Han, J.-S., Lee, K.-R., Kozukue, N., (2006) 'Distribution of catechins, theaflavins, caffeine, and theobromine in 77 teas consumed in the United States', J. Food Sci. 70, pp. C550–C559.

Guimarães, R., Barros, L., Dueñas, M., Calhelha, R.C., Carvalho, A.M., Santos-Buelga, C., Queiroz, M.J.R.P., Ferreira, I.C.F.R., (2013) Nutrients, phytochemicals and bioactivity of wild Roman chamomile: A comparison between the herb and its preparations. Food Chem. 136, pp. 718–725

Haghi, G., (2014) 'Analysis of phenolic compounds in Matricaria chamomilla and its extracts by UPLC-UV', Res Pharm Sci, 9(1), pp. 31-7

Hanrahan, J.R., Chebib, M., Johnston, G.A.R., (2015) 'Interactions of flavonoids with ionotropic GABA receptors, Advances in Pharmacology, Elsevier, pp. 189–200

Hanrahan, J.R., Chebib, M., Johnston, G.A.R., (2011. Flavonoid modulation of GABAA receptors: Flavonoids and GABAA receptors. Br. J. Pharmacol. 163, 234–245

Harbourne, N., Jacquier, J.C., O’Riordan, D., (2009) 'Optimisation of the extraction and processing conditions of chamomile (Matricaria chamomilla L.) for incorporation into a beverage', Food Chem, 115, pp. 15–19

Head, K.A., Kelly, G.S., (2009) 'Nutrients and botanicals for treatment of stress: adrenal fatigue, neurotransmitter imbalance, anxiety, and restless', Sleep,14, pp. 27

Hood, S.D., Norman, A., Hince, D.A., Melichar, J.K., Hulse, G.K., (2014) Benzodiazepine dependence and its treatment with low dose flumazenil: Benzodiazepine dependence and its treatment. Br. J. Clin. Pharmacol. 77, 285–294

Johnston, G., (2005) 'GABAA receptor channel pharmacology', Curr. Pharm. Des. 11, pp. 1867–1885

Kawiji, K., Anam, C., Manuhara, G.J., Fakhrudin, M.I., (2009) 'Kajian karakteristik oleoresin jahe berdasarkan ukuran dan lama perendaman serbuk jahe dalam etanol', Caraka tani, J. Sustain. Agric. 24, pp. 61

Kumar, D., Bhat, Z.A., (2012) 'Anti-anxiety activity of methanolic extracts of different parts of Angelica archangelica Linn', J. Tradit. Complement. Med., 2, pp. 235–241

Lara, D.R., (2010) Caffeine, mental health, and psychiatric disorders, J. Alzheimers Dis, 20, pp. S239–S248

López-Cruz, L., Pardo, M., Salamone, J.D., Correa, M., (2014) 'Differences between the nonselective adenosine receptor antagonists caffeine and theophylline in motor and mood effects: Studies using medium to high doses in animal models', Behav. Brain Res, 270, pp. 213–222.

López-Cruz, L., Salamone, J.D., Correa, M., (2013) 'The Impact of caffeine on the behavioral effects of ethanol related to abuse and addiction: a review of animal studies', J. Caffeine Res. 3, pp. 9–21

Manber, R., Edinger, J.D., Gress, J.L., Pedro-Salcedo, M.G.S., Kuo, T.F., Kalista, T., (2008) 'Cognitive behavioral therapy for insomnia enhances depression outcome in patients with comorbid major depressive disorder and insomnia', Sleep, 31, pp. 489–495

Marwa, C., Fikri-Benbrahim, K., Ou-Yahia, D., Farah, A., (2017) 'African peppermint (Mentha piperita) from Morocco: Chemical composition and antimicrobial properties of essential oil', 8(3), pp. 86–90

McAlpine, M., Ward, W., (2016) 'Influence of steep time on polyphenol content and antioxidant capacity of black, green, rooibos, and herbal teas' Beverages 2, pp. 17

Mendonça Netto, S., Warela, R.W.B., Fechine, M.F., Queiroga, M.N., Quintans-Júnior, L.J., (2009) 'Anxiolytic-like effect of Rauvolfia ligustrina Willd: ex Roem. & Schult., Apocynaceae, in the elevated plus-maze and hole-board tests. Rev', Bras. Farmacogn, 19, 888–892

Morgan, P.T., Pace-Schott, E.F., Mason, G.F., Forselius, E., Fasula, M., Valentine, G.W., Sanacora, G., (2012) 'Cortical GABA levels in primary insomnia', Sleep, 35, pp. 807–814

Morin, C.M., Rodrigue, S., Ivers, H., (2003) Role of Stress, Arousal, and Coping Skills in Primary Insomnia: Psychosom. Med. 65, pp. 259–267

Motivala, S.J., Irwin, M.R., (2007) 'Sleep and Immunity: cytokine pathways linking sleep and health outcomes', Curr. Dir. Psychol. Sci. 16, pp. 21–25

Muchuweti, M., Kativu, E., Mupure, C.H., Chidewe, C., Ndhlala, A.R., Benhura, M.A.N., 2007. Phenolic Composition and Antioxidant Properties of Some Spices. Am. J. Food Technol. 2, 414–420.

Mullington, J.M., Haack, M., Toth, M., Serrador, J.M., Meier-Ewert, H.K., (2009) 'Cardiovascular, inflammatory, and metabolic consequences of sleep deprivation', Prog. Cardiovasc. Dis, 51, pp. 294–302

Olayiwola, G., Obafemi, C.A., Taiwo, F.O., (2007) Synthesis and neuropharmacological activity of some quinoxalinone derivatives, Afr J Biotechnol , 6(6), pp. 777-786

Opp, M.R., (2005) 'Cytokines and sleep', Sleep Med. Rev. 9, pp. 355–364

Palma, B.D., Gabriel, A., Colugnati, F.A.B., Tufik, S., (2006) 'Effects of sleep deprivation on the development of autoimmune disease in an experimental model of systemic lupus erythematosus', Am. J. Physiol.-Regul. Integr. Comp. Physiol, 291, pp. R1527–R1532

Pasakawee, K., Utama-ang, N., (2018) 'Antioxidant activity and starch-digesting enzyme inhibition of selected Thai herb extracts Chiang Mai', J. Sci , 5(1), pp. 263-276

Paterson, L.M., Wilson, S.J., Nutt, D.J., Hutson, P.H., Ivarsson, M., (2007) 'A translational, caffeine-induced model of onset insomnia in rats and healthy volunteers' Psychopharmacology (Berl.), 191, pp. 943–950

Patil, S.P., Liu, C., Alban, J., Yang, N., Li, X.-M., (2014) 'Glycyrrhiza uralensis flavonoids inhibit brain microglial cell TNF-α secretion, p-IκB expression, and increase brain-derived neurotropic factor (BDNF) secretion' J. Tradit. Chin. Med. Sci. 1, 28–37. https://doi.org/10.1016/j.jtcms.2014.11.004

Pearson, N.J., (2006) 'Insomnia, trouble sleeping, and complementary and alternative medicine: analysis of the 2002 National Health Interview Survey Data. Arch. Intern. Med, 166, pp.1775

Pham, X., Sun, C., Chen, X., van den Oord, E.J.C.G., Neale, M.C., Kendler, K.S., Hettema, J.M., (2009) 'Association study between GABA receptor genes and anxiety spectrum disorders', Depress. Anxiety 26, pp. 998–1003

Raini, M., (2011) Kajian: Khasiat dan keamanan stevia sebagai pemanis pengganti gula, 21 (4). [In Indonesian]

Ramlah, (2017) 'Penentuan suhu dan waktu optimum penyeduhan daun teh hijau (Camellia sinensis L) P+2 Terhadap Kandungan Antioksidan Kafein, Tanin dan Katekin', Minor Thesis. Fakultas Sains dan Teknologi UIN Alauddin Makassar. [In Indonesian]

Rostagno, M.A., Manchón, N., D’Arrigo, M., Guillamón, E., Villares, A., García-Lafuente, A., Ramos, A., Martínez, J.A., (2011) 'Fast and simultaneous determination of phenolic compounds and caffeine in teas, mate, instant coffee, soft drink and energetic drink by high-performance liquid chromatography using a fused-core column' Anal. Chim. Acta, 685, pp. 204–211

Roth, T., Roehrs, T., (2003) 'Insomnia: Epidemiology, characteristics, and consequences', Clin. Cornerstone ,5, pp. 5–15

Roth, T., Roehrs, T., Pies, R., (2007) 'Insomnia: Pathophysiology and implications for treatment', Sleep Med. Rev., 11, pp. 71–79

Rustay, N.R., Wahlsten, D., Crabbe, J.C., (2003) 'Influence of task parameters on rotarod performance and sensitivity to ethanol in mice', Behav. Brain Res. 141, pp. 237–249

Sadeghniiat-Haghighi, K., Montazeri, A., Khajeh-Mehrizi, A., Nedjat, S., Aminian, O., (2014). 'The Insomnia Severity Index: cross-cultural adaptation and psychometric evaluation of a Persian version'. Qual. Life Res, 23, pp. 533–537

Saniah, K., Hasimah, H.A., (2008) 'Development of Morinda citrifolia citrus-flavoured drink using Response Surface Methodology (RSM), 36(1), pp. 87-89

Sharafzadeh, S., Alizadeh, O., n.d. German and Roman Chamomile. Journal of Applied Pharmaceutical Sciences, 1(10), pp. 1-5

Shinomiya, K., Inoue, T., Utsu, Y., Tokunaga, S., Masuoka, T., Ohmori, A., Kamei, C., 2005. Hypnotic Activities of Chamomile and Passiflora Extracts in Sleep-Disturbed Rats. Biological and Pharmaceutical Bulletin. 28, 808–810

Silveira, L.F.L. da, Bocakova, M., Mermudes, J.R.M., 2015. Taxonomy of the Brazilian firefly Cladodes illigeri (Coleoptera, Lampyridae): morphology and new records', Iheringia Série Zoololgy, 105, pp. 359–366

Stein, M., Stein, D., (2008) 'Social anxiety disorder', Lancet, 29 (371), pp. 115-25

Stojanović, N.M., Samardžić, L., Randjelović, P.J., Radulović, N.S., (2017) 'Prevalence of self-medication practice with herbal products among non-psychotic psychiatric patients from southeastern Serbia: A cross-sectional study', Saudi Pharmaceutical Journal, 25(6), pp. 884–890

Sufiana, H. (2014) 'Uji Aktivitas Antioksidan Dan Sitotoksisitas Campuran Ekstrak Metanol Kayu Sepang (Caesalpinia sappan L.) Dan Kulit Kayu Manis (Cinnamomum burmannii B.)', J. Kim. Khatulistiwa, 3, pp. 50–55. [In Indonesian]

Sultana, T., Mannan, M.A., Ahmed, T., 2018. Evaluation of central nervous system (CNS) depressant activity of methanolic extract of Commelina diffusa Burm. in mice', Clin. Phytoscience 4

Tfouni, S.A.V., Camara, M.M., Kamikata, K., Gomes, F.M.L., Furlani, R.P.Z. (2018) 'Caffeine in teas: levels, transference to infusion and estimated intake, Food Science Technology, 38, pp. 661–666

Thomas, M. (2003) 'Neural basis of alertness and cognitive performance impairments during sleepiness II. Effects of 48 and 72 h of sleep deprivation on waking human regional brain activity. Thalamus & Related System, 2(3), pp. 199–229

Tim Hortons Research and Development, (2017) Caffeine Content


Vallverdú-Queralt, A., Regueiro, J., Martínez-Huélamo, M., Rinaldi Alvarenga, J.F., Leal, L.N., Lamuela-Raventos, R.M., (2014) 'A comprehensive study on the phenolic profile of widely used culinary herbs and spices: Rosemary, thyme, oregano, cinnamon, cumin and bay', Food Chem, 154, pp. 299–307

Vgontzas, A.N., Papanicolaou, D.A., Bixler, E.O., Hopper, K., Lotsikas, A., Lin, H.-M., Kales, A., Chrousos, G.P., (2000) 'Sleep Apnea and Daytime Sleepiness and Fatigue: Relation to Visceral Obesity, Insulin Resistance, and Hypercytokinemia', Sleep Apnea, 85, pp. 8

Wuryantoro, H., Susanto, W.H., (2014) 'Building Standard Operating Procedures for Home Industry of Instant Sweetener of Stevia Extract (Stevia rebaudiana)', Jurnal Pangan Dan Agroindustri, 2, pp. 76–87 [In Indonesian]

Zekovic, Z., Cvetanovic, A., Pavlic, B., Svarc-Gajic, J., Radojkovic, M., (2015) 'The optimization of the extraction process of flavonoids from fermented chamomile ligulate flowers. Savrem. Tehnol', 4, pp. 54–63

DOI: https://doi.org/10.21776/ub.jtp.2019.002.02.2


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