Determination of Caffeic Acid in Ethanolic Extract of Spent Coffee Grounds by High-Performance Liquid Chromatography with UV Detection

Michael Raharja Gani(1*), Enade Perdana Istyastono(2)

(1) Faculty of Pharmacy, Sanata Dharma University, Campus III Paingan, Maguwoharjo, Depok, Sleman, Yogyakarta 55282, Indonesia
(2) Faculty of Pharmacy, Sanata Dharma University, Campus III Paingan, Maguwoharjo, Depok, Sleman, Yogyakarta 55282, Indonesia
(*) Corresponding Author


We developed a method for determining the caffeic acid in spent coffee grounds. The spent coffee ground solution was prepared by blending 3 g spent coffee grounds with 60 mL ethanol/water (40/60 v/v) for 2 h on a hot plate magnetic stirrer (60 °C, 350 rpm). The mixture was filtered and the filtrate was concentrated under vacuum (60 °C) to 5 mL. The method employed a reversed-phase high-performance liquid chromatography with a UV detector. We used a Phenomenex Luna column (250 × 4.6 mm; i.d., 5 µm) under isocratic elution, and the mobile phase was acetonitrile-methanol-aqueous formic acid (10:10:80 v/v), with a flow rate of 0.9 mL/min. Analysis was performed at 324 nm. The column temperature was set at 27 °C temperature. The results showed that this method was selective for quantifying the caffeic acid in spent coffee grounds with good linearity in the range of 1.31–17.07 μg/mL. The detection and quantitation limits were 0.28 and 0.84 μg/mL, respectively. The mean intraday and interday recoveries were 83.80–95.17% and 82.16–97.40%, respectively. Intraday and interday precision expressed as the relative standard deviation (RSD) were below 7.3%. There was 0.17% ± 0.006 w/w caffeic acid in the spent coffee grounds (RSD = 3.63%, n = 3).


caffeic acid; reversed phase high performance liquid chromatography; spent coffee grounds

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[1] Anonymous, 2014, Indonesia’s Coffee Industry Needs Growth Capital, agriculture/article/2014/indonesia_s_coffee_industry_needs_growth_capital.php, accessed on 19 November 2020.

[2] Donald, M.G., and Rahmanulloh, A., 2019, Indonesia Coffee Annual Report 2019, https://apps.fas.usda. gov/newgainapi/api/report/downloadreportbyfilename?filename=Coffee%20Annual_Jakarta_Indonesia_5-15-2019.pdf, accessed on 20 November 2020.

[3] Cruz, R., Cardoso, M.M., Fernandes, L., Oliveira, M., Mendes, E., Baptista, P., Morais, S., and Casal, S., 2012, Espresso coffee residues: A valuable source of unextracted compounds, J. Agric. Food. Chem., 60 (32), 7777–7784.

[4] Jenkins, R.W., Stageman, N.E., Fortune, C.M., and Chuck, C.J., 2014, Effect of the type of bean, processing, and geographical location on the biodiesel produced from waste coffee grounds, Energy Fuels, 28 (2), 1166–1174.

[5] Anonymous, 2020, Trade Statistics-December 2019, International Coffee Organization, http://www.ico. org, accessed on 19 November 2020.

[6] Mata, T.M., Martins, A.A., and Caetano, N.S., 2018, Bio-refinery approach for spent coffee grounds valorization, Bioresour. Technol., 247, 1077–1084.

[7] Murthy, P.S., and Naidu, M.M., 2012, Sustainable management of coffee industry by-products and value addition–A review, Resour. Conserv. Recycl., 66, 45–58.

[8] Ballesteros, L.F., Teixeira, J.A., and Mussatto, S.I., 2014, Chemical, functional, and structural properties of spent coffee grounds and coffee silverskin, Food Bioprocess Technol., 7 (12), 3493–3503.

[9] Aprilia, A.A., 2013, Antimicrobial and Antioxidant Activities of Microwave Assisted Extracts from Coffee Ground Residue in Chiang Rai Province, Thailand, Undergraduate Thesis, Bogor Agricultural University, Bogor.

[10] Anshari, M.S., 2018, Penentuan Senyawa Kafein dan Total Fenol pada Ekstrak Ampas Kopi Robusta (Coffea canephora) dengan Metode Ekstraksi Microwave Assisted, Sokletasi, dan Maserasi serta Uji Aktifitas Antibakteri Staphylococcus aureus, Undergraduate Thesis, Universitas Islam Indonesia, Yogyakarta.

[11] Naidhu, M.M., Sulochanamma, G., Sampathu, S.R., and Srinivas, P., 2008, Studies on extraction and antioxidant potential of green coffee, Food Chem., 107 (1), 377–384.

[12] Huang, M.T., and Ferraro, T., 1992, Phenolic compounds in food and cancer prevention, ACS Symp. Ser., 507, 8–34.

[13] Chiang, E.P.I., Tsai, S.Y., Kuo, Y.H., Pai, M.H., Chiu, H.L., Rodriguez, R.L., and Tang, F.Y., 2014, Caffeic acid derivatives inhibit the growth of colon cancer: Involvement of the PI3-K/Akt and AMPK signaling pathways, PLoS ONE, 9 (6), e99631.

[14] Silva, H., and Lopes, N.M.F., 2020, Cardiovascular effects of caffeic acid and its derivatives: A comprehensive review, Front. Physiol., 11, 595516.

[15] Shi, Z.H., Li, N.G., Shi, Q.P., Tang, H., Tang, Y.P., Li, W., Yin, L., Yang, J.P., and Duan, J.A., 2013, Synthesis and structure-activity relationship analysis of caffeic acid amides as selective matrix metalloproteinase inhibitors, Bioorg. Med. Chem. Lett., 23 (5), 1206–1211.

[16] Mendes, V., Mendes, S., Gomes, B., Anjos, O., and Campos, M.G., 2015, Caffeic acid derivates in spent coffee ground as potential crude material for drug discovery, Planta Med., 81, 172.

[17] Ramón-Gonçalves, M., Gómez-Mejía, E., Rosales-Conrado, N., León-González, M.G., and Madrid, Y., 2019, Extraction, identification and quantification of polyphenols from spent coffee grounds by chromatographic methods and chemometric analyses, Waste Manage., 96, 15–24.

[18] Angeloni, S., Nzekoue, F.K., Navarini L, Sagratini, G., Torregiani, E., Vittori, S., and Caprioli, G., 2020, An analytical method for the simultaneous quantification of 30 bioactive compounds in spent coffee ground by HPLC-MS/MS, J. Mass Spectrom., 55 (11), e4519.

[19] Anonymous, 2017, Differences between Arabica and Robusta Coffee,, accessed on 30 March 2021.

[20] Anonymous, 2020, Advantages and Disadvantages of Gradient Elution in HPLC,, accessed on 30 March 2021.

[21] Doncea, S.M., Stoica, R., Mariana-Rodica, I., Inna, T., and Valeriu, P., 2010, An HPLC Method for Identification and Separation of some Phenolic Acids in the Coffee, Petroleum - Gas University of Ploiesti Bulletin, Technical Series, 62 (3A), 143–148.

[22] Andrade, P.B., Leitão, R., Seabra, R.M., Oliveira, M.B., and Ferreira, M., 1997, Development of an HPLC/diode-array detector method for simultaneous determination of seven hydroxy-cinnamic acids in green coffee, J. Liq. Chromatogr. Relat. Technol., 20 (13), 2023–2030.

[23] Juliantari, N.P.D., Wrasiati, L.P., dan Wartini, N.M., 2018, Karakteristik ekstrak ampas kopi bubuk robusta (Coffea canephora) pada perlakuan konsentrasi pelarut etanol dan suhu maserasi, JRMA, 6 (3), 243–249.

[24] International Conference on Harmonization (ICH), 1995, Q2 (R1): Validation of analytical procedures: Text and methodology – Step 5, CPMP/ICH/381/95, European Medicines Agency, Canary Wharf, London.

[25] AOAC, 2016, Official Methods of Analysis of AOAC International, 20th Ed., Eds. Latimer, G.W., AOAC International, Rockville, Maryland, USA, 8–9.


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