Quantification of rutin, quercitrin and quercetin in Cosmos caudatus Kunth by reverse phase high performance liquid chromatography

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M.M.A. Sharifuldin
Z. Ismail
A.F.A. Aisha
E.K. Seow
H.K. Beh


medicinal plant, extracts, method validation, marker compounds, quality control assurance


Cosmos caudatus is a traditional medicinal plant from the family Compositae. The fresh leaves are consumed as appetizer and for health benefits such as antioxidant activity, to improve blood circulation and to promote the formation of healthy bones. Our study aims to develop a reverse phase high performance liquid chromatography method for quantification of rutin, quercitrin and quercetin in C. caudatus extracts. The method was validated for selectivity, linearity, precision, accuracy and limits of detection and quantification. Selectivity was confirmed based on retention time and ultraviolet-visible spectra; linearity was in the range 0.5-500 ?g/ml at R2=1.0; the intraday and interday precision was determined as the relative standard deviation of peak area and retention time. Percentage recovery, limits of detection and the limits of quantification of rutin, quercitrin and quercetin were also determined. The reported method was applied for the quantification of the 3 marker compounds in 15 extracts of C. caudatus. Results showed quercitrin as the major constituent in C. caudatus; hence it may be used as a reference compound for standardisation of this medicinal herb. The method can be considered as analytical tool for quality control, stability studies, pharmacokinetics, and standardisation purposes.

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Abas, F., 2005. Phytochemical and biological activity studies of Cosmos caudatus and Curcuma mangga and the online characterization of bioactive fractions from Melicope ptelefolia. PhD thesis, University of Putra, Malaysia.
Abas, F., Lajis, N.H., Israf, D.A., Khozirah, S. and Umi Kalsom, Y., 2006. Antioxidant and nitric oxide inhibition activities of selected Malay traditional vegetables. Food Chemistry95: 566-573.
Abas, F., Shaari, K., Lajis, N.H., Israf, D.A. and Kalsom, Y.U., 2003. Antioxidative and radical scavenging properties of the constituents isolated from Cosmos caudatus Kunth. Natural Product Sciences 9(4): 245-248.
Aisha, A.F.A., Abu-Salah, K.M., Siddiqui, M.J., Ismail, Z. and Majid, A.M.S.A., 2012. Quantification of ?-, ?- and ?-mangostin in Garcinia mangostana fruit rind extracts by a reverse phase high performance liquid chromatography. Journal of Medicinal Plants Research 6: 4526-4534.
Andarwulan, N., Batari, R., Sandrasari, D.A., Bolling, B. and Wijaya, H., 2010. Flavonoid content and antioxidant activity of vegetables from Indonesia. Food Chemistry 121: 1231-1235.
Boligon, A.A., Agertt, V., Janovik, V., Cruz, R.C., Campos, M.M.A., Guillaume, D., Athayde, M.L. and Dos Santos, A.R.S., 2012. Antimycobacterial activity of the fractions and compounds from Scutia buxifolia. Brazilian Journal of Pharmacognosy 22: 45-52.
Cavalier, E., Rozet, E., Dubois, N., Charlier, C., Hubert, P., Chapelle, J.P., Krzesinski, J.M. and Delanaye, P., 2008. Performance of iohexol determination in serum and urine by HPLC: Validation, risk and uncertainty assessment. Clinica Chimica Acta 396: 80-85.
Chen, Y., Yan, Y., Xie, M.Y., Nie, S.P., Liu, W., Gong, X.F. and Wang, Y.X., 2008. Development of a chromatographic fingerprint for the chloroform extracts of Ganoderma lucidum by HPLC and LC-MS. Journal of Pharmaceutical and Biomedical Analysis 47: 469-477.
Dubber, M.-J. and Kanfer, I., 2004. High-performance liquid chromatographic determination of selected flavonols in Ginkgo biloba solid oral dosage forms. Journal of Pharmaceutical Sciences 7(3): 303-309.
Huda-Faujan, N., Noriham, A., Norakkiah, A.S. and Babji, A.S., 2007. Antioxidative activities of waters extracts of some Malaysian herbs. ASEAN Food Journal 14: 61-68.
Huda-Faujan, N., Noriham, A., Norrakiah, A.S. and Babji, A.S., 2009. Antioxidant activity of plants methanolic extracts containing phenolic compounds. African Journal of Biotechnology8(3): 484-489.
International Conference on Harmonisation (ICH), 1997. Guidance for industry, q2b: validation of analytical procedures: methodology. ICH, USA. Available at: http://tinyurl.com/jzraea3.
Jadhav, R.B., Patil, C.R., Bhpoe, S. and Murumkar, C.V., 2003. Herbal drug standardization and quality assurance of raw materials: a rational approach. Natural Product Radiance 2: 134-140.
Leong, L.P. and Shui, G., 2001. An investigation of antioxidant capacity of fruits in Singapore markets. Food Chemistry 76: 69-75.
Li, J., Jiang, B., Liu, X., Zhang, J., Chen, X. and Bi, K., 2007. Simultaneous determination of five bioactive flavonoids in Hypericum japonicum Thunb by High-performance Liquid Chromatography. Asian Journal of Traditional Medicine 2: 75-81.
Marin, A., Garcia, E., Garcia, A. and Barbas, C., 2002. Validation of a HPLC quantification of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations: capsules and sachets. Journal of Pharmaceutical and Biomedical Analysis 29: 701-714.
Mediani, A., Abas, F., Tan, C.P., Khatib, A. and Nordin, H.L., 2012. Influence of growth stage and season on the antioxidant constituents of Cosmos caudatus. Plant Foods for Human Nutrition 67: 344-350.
Mendonça-Filho, R.R., 2006. Bioactive phytocompounds: new approaches in the photo sciences. In: Ahmad, I., Aqil, F. and Owais, M. (eds.) Modern Phytomedicine. Wiley-VCH, Weinheim, Germany.
Mosihuzzaman, M. and Choudhary, M.I., 2008. Protocols on safety, efficacy, standardization, and documentation of herbal medicine. Pure and Applied Chemistry 80: 2195-2230.
Mustafa, R.A., Hamid, A.A., Mohamed, S. and Bakar, F.A., 2010. Total phenolic compounds, flavonoids, and radical scavenging activity of 21 selected tropical plants. Journal of Food Science75: 28-35.
Norazlina, M., Ehsan, S.Z., Adilah, N.K., Lee, C.P., Farhana, E., Derick, P., Ima-Nirwana, S., Nazrun, A.S. and Norliza, M., 2013. Acute toxicity study of Cosmos caudatus on biochemical parameters in male rats. Sains Malaysia 42(9): 1247-1251.
Shui, G., Leong, L.P. and Wong, S.P., 2005. Rapid screening and characterization of antioxidants of Cosmos caudatus using liquid chromatography coupled with mass spectrometry. Journal of Chromatography B 827: 127-138.
Takami, S., Imai, T., Hasumura, M., Cho, Y.M., Onose, J. and Hirose, M., 2008. Evaluation of toxicity of green tea catechins with 90-day dietary administration to F344 rats. Food and Chemical Toxicology 46: 2224-2229.
Wong, S.P., Leong, L.P. and William Koh, J.H., 2006. Antioxidant activities of aqueous extracts of selected plants. Food Chemistry99: 775-783.