Author(s): Sarita Garg, Rubal Chahal, Deepak Kaushik, Rakesh Kumar, Vineet Mittal

Email(s): drvineet.pharma@mdurohtak.ac.in

DOI: 10.52711/0974-360X.2023.00383   

Address: Sarita Garg1, Rubal Chahal1, Deepak Kaushik1, Rakesh Kumar2, Vineet Mittal1*
1Department of Pharmaceutical Sciences, Maharshi Dayanand University, Rohtak, Haryana, India.
2Department of Pharmacy, Vaish Institute of Pharmaceutical Education and Research, Rohtak, India.
*Corresponding Author

Published In:   Volume - 16,      Issue - 5,     Year - 2023


ABSTRACT:
Mulberry, which pertains to the Moraceae family and the genus Morus,has been used in medicine for ages because of its chemical composition and pharmacological action. The plant is recognized in traditional Chinese medicine for its antiphlogistic, diuretic, expectorant, and antidiabetic characteristics. The mulberry leaf is high in flavonoids, which have a variety of biological functions, including antioxidant potential. Quercetin, isoquercetin, rutin, isoquercitrin, quercitrin, luteolin, chlorogenic acid, and other flavonoids can be found in their leaves. The major constituents of total flavones in mulberry leaf extract are rutin and quercetin. The goal of the RP-HPLC approach was to design and validate a method for identifying these elements. HPLC (Shimadzu Technologies LC series) with UV-visible detector at 259nm and C18 column (250mm 4.6mm, 5µm particle size) was used to achieve chromatographic separation (Phenomenex Luna). Solvents (acetonitrile and 0.1 percent v/v solution of glacial acetic acid) were used in the isocratic elution mode. The run time was 10 minutes. The column temperature was 37 degrees Celsius with sample insertion volume 20µl and the flow velocity was 1ml/min. For quercetin and rutin, the devised method produced strong linearity, high accuracy, high precision, high repeatability, high robustness, and high ruggedness. For quercetin and rutin, the LOD and LOQs values were determined to be within acceptable levels.


Cite this article:
Sarita Garg, Rubal Chahal, Deepak Kaushik, Rakesh Kumar, Vineet Mittal. RP-HPLC Method Development and Validation for Simultaneous Estimation of Rutin and Quercetin in Morus alba L. leaf extract. Research Journal of Pharmacy and Technology 2023; 16(5):2327-5. doi: 10.52711/0974-360X.2023.00383

Cite(Electronic):
Sarita Garg, Rubal Chahal, Deepak Kaushik, Rakesh Kumar, Vineet Mittal. RP-HPLC Method Development and Validation for Simultaneous Estimation of Rutin and Quercetin in Morus alba L. leaf extract. Research Journal of Pharmacy and Technology 2023; 16(5):2327-5. doi: 10.52711/0974-360X.2023.00383   Available on: https://rjptonline.org/AbstractView.aspx?PID=2023-16-5-42


REFERENCES:
1.    Sangeeta Sankhalkar and VrundaVernekar (2017). Quantitative and Qualitative Analysis of Phenolic and Flavonoid Content in Moringa oleifera Lam and Ocimumtenuiflorum L., Pharmacognosy Res. 2016; 8(1): 16–21.
2.    Hafez-Taghva, Pardis, Zamzad, Mina, Khalafi, Lida.Total flavonoid content and essential oil composition of Chaenomeles japonica (Thunb.) Lindl. ex Spach from North of Iran. Indian Journal of Natural Products and Resources. 2016;7 (1):90-92.
3.    Ramesh HL, Sivaram V and Murthy VNY (2014).  Antioxidant and medicinal properties of mulberry (morus sp.): A review. Wor. J. Pharm. Res. 2014; 3(6): 320-343.
4.    Chan, E.W., Lye, P.Y. and Wong, S.K.. Phytochemistry, pharmacology, and clinical trials of Morusalba. Chinese Journal of Natural Medicines. 2016;14: 17-30.
5.    Wu, S.L., Yang, X.B., Liu, L.Q., Jiang, T., Wu, H., Su, C., Qian, Y.H. and Jiao, F. Agrobacterium-mediated transient MaFT expression in mulberry (MorusalbaL.) leaves. Bioscience, Biotechnology, and Biochemistry. 2015; 79: 1266-1271.
6.    Ercisil S, Orhan E. Chemical composition of white (Morusrubra) and black (Morusnigra) mulberry fruits. Food Chem. 2007; 103:1380–84.
7.    Faiz-ul Hassan, Muhammad Adeel Arshad, Mengwei Li, Muhammad Saif-ur Rehman, Juan J. Loor and Jiaxiang Huang. Potential of Mulberry Leaf Biomass and Its Flavonoids to Improve Production and Health in Ruminants: Mechanistic Insights and Prospects, Animals. 2020; 10, 2076; doi:10.3390/ani10112076.
8.    Nomura T. Phenolic compounds of the mulberry tree and related plants. In:Herz W, Grisebach H, Kirby GW, Tamm CH editors. Progress in the Chemistry of Organic Natural Products. 1988; 53;87-201.
9.    Varghese SM, Thomas J. Polyphenolic constituents in mulberry leaf extract (M. latifolia L. cv. BC259) and its antidiabetic effect in streptozotocin induced diabetic rats. Pak J Pharm Sci. 2019; 32(1):69-74.
10.    Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999; 64:555–9.
11.    Bahare Salehi, Athar Ata, Nanjangud V. Anil Kumar, FarukhSharopov, Karina Ramírez-Alarcón, Ana Ruiz-Ortega, SeyedAbdulmajidAyatollahi. Antidiabetic Potential of Medicinal Plants and Their Active Components. Biomolecules. 2019, 9, 551; doi:10.3390/biom9100551.
12.    Singab AN, El–Beshbishy HA, Yonekawa M, Nomura T, Fukai T. Hypoglycemic effect of Egyptian Morus alba root bark extract: effect on diabetes and lipid peroxidation of streptozotocin-induced diabetic rats. J Ethnopharmacol. 2005; 100(3):333–338.
13.    Chae JY, Lee JY, Hoang IS, Whangbo D, Choi PW, Lee WC, Kim JW, Kim SY, Choi SW, Rhee SJ. (2003). Analysis of functional components of leaves of different mulberry cultivars. J Kor Soc Food Sci Nutr. 2003; 32(1):15–21.
14.    E. Park, S. M. Lee, J. e. Lee, and J. H. Kim, “Anti-inflammatory activity of mulberry leaf extract through inhibition of NF-κB,” Journal of Functional Foods. 2013; 5(1); 178–186.
15.    J. Naowaboot, P. Pannangpetch, V. Kukongviriyapan, B. Kongyingyoes, and U. kukongviriyapan, “Antihyperglycemic, antioxidant and antiglycation activities of mulberry leaf extract in streptozotocin-induced chronic diabetic rats,” Plant Foods for Human Nutrition. 2009; 64(2)116–121, 2009.
16.    N. C. Yang, K. Y. Jhou, and C. Y. Tseng. Antihypertensive effect of mulberry leaf aqueous extract containing γaminobutyric acid in spontaneously hypertensive rats. Food Chemistry. 2012; 132 (4); 1796–1801.
17.    Gryn-Rynko, A., Bazylak, G. and Olszewska-Slonina, D. New potential phytotherapeutics obtained from white mulberry (MorusalbaL.) leaves. Biomedicine and Pharmacotherapy. 2016; 84: 628-636.
18.    Wilson, R.D. and Islam, M.S. Effects of white mulberry (Morusalba) leaf tea investigated in a type 2 diabetes model of rats. Acta Poloniae Pharmaceutica. 2015; 72: 153-160.
19.    Zhou, Z., Zhou, B., Ren, L. and Meng, Q. Effect of ensiled mulberry leaves and sun-dried mulberry fruit pomace on finishing steer growth performance, blood biochemical parameters, and carcass characteristics. PLoSOne. 2014; 9: e85406.
20.    R. K. Datta, A. Sarkar, P. R. M. Rao, and N. R. Singhvi. Utilization of mulberry as animal fodder in India. in Mulberry for Animal Production, M. D. S´anchez, Ed.2002; 183–188.
21.    Deepa, M., Sureshkumar, T., Satheeshkumar, P.K. and Priya, S. Antioxidant rich Morus alba leaf extract induces apoptosis in human colon and breast cancer cells by the downregulation of nitric oxide produced by inducible nitric oxide synthase. Nutrition and Cancer . 2013; 65: 305-310.
22.    Fallah, S., Karimi, A., Panahi, G., GerayeshNejad, S., Fadaei, R. and Seifi, M. Human colon cancer HT-29 cell death responses to doxorubicin and Morus alba leaves flavonoid extract. Cellular and Molecular Biology (Noisy-le-grand). 2016; 62: 72-77.
23.    Naowaratwattana, W., De-Eknamkul, W. and De Mejia, E.G. Phenolic-containing organic extracts of mulberry (Morus alba L.) leaves inhibit HepG2 hepatoma cells through G2/M phase arrest, induction of apoptosis, and inhibition of topoisomerase II alpha activity. Journal of Medicinal Food. 2010; 13: 1045-1056.
24.    Ju WT, Kwon OC, Kim HB, Sung GB, Kim HW, Kim YS. Qualitative and quantitative analysis of flavonoids from 12 species of Korean mulberry leaves. J Food Sci Technol. 2018;55(5):1789-1796. doi:10.1007/s13197-018-3093-2
25.    T. Katsube, N. Imawaka, Y. Kawano, Y. Yamazaki, K. Shiwaku, and Y. Yamane. Antioxidant flavonol glycosides in mulberry (Morus alba L.) leaves isolated based on LDL antioxidant activity. Food Chemistry. 2006; 97(1): 25–31.
26.    N. Nuengchamnong, K. Ingkaninan, W. Kaewruang, S. Wongareonwanakij, and B. Hongthongdaeng. Quantitative determination of 1-deoxynojirimycin in mulberry leaves using liquid chromatography-tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis. 2007; 44(4): 853–858.
27.    Eric Wei-Chiang CHAN, Phui-Yan LYE, Siu-Kuin WONG. Phytochemistry, pharmacology, and clinical trials of Morus alba [J]. Chinese Journal of Natural Medicines. 2016; 14(1): 17-30.
28.    Kim GN, Jang HD. Flavonol content in the water extracts of the mulberry (Morusalba L.) leaf and their antioxidant capacities. J Food Sci. 2011; 76(6): C869-C873. doi:10.1111/j.1750-3841.2011.02262.x
29.    P. Pothinuch and S. Tongchitpakdee. Melatonin contents in mulberry (Morusspp.) leaves: effects of sample preparation, cultivar, leaf age and tea processing. Food Chemistry. 2011; 128(2): 415–419.
30.    Pothinuch P, Tongchitpakdee S. Phenolic analysis for classification of mulberry (Morus spp.) leaves according to cultivar and leaf age. Journal of Food Quality. 2019; 1-11. https://doi.org/10.1155/2019/2807690
31.    Gulab Khan Rohela, Pawan Shukla, Muttanna, Rajesh Kumar, Sukhen Roy Chowdhury. Mulberry (Morus spp.): An ideal plant for sustainable development, Trees, Forests and People. 2020; 2 : 100011.
32.    Sultana B, Anwar F. Flavonols (Kaempeferol, quercetin, myricetin) contents of selected fruits, vegetables and medicinal plants. Food Chem. 2008; 108: 879–884.
33.    Ou-yang Z, Caoa X, Weia Zhen Ou-yang Y, Zhanga WWY, Zhao M, Duan J. Pharmacokinetic study of rutin and quercetin in rats after oral administration of total flavones of mulberry leaf extract. Revista Brasileira de Farmacognosia. 2013; 23(5):776-782.
34.    Zhai KF, Duan H, Shi SX, Liu LL, Cao WG, Gao GZ, Shan LL. Synchronised determination of chlorogenic acid and five flavonoids in mulberry leaves using HPLC with photodiode array detection. Quality Assurance and Safety of Crops and Foods. 2018;10(2):175-82.
35.    Pothinuch P, Tongchitpakdee S. Phenolic analysis for classification of mulberry (Morus spp.) leaves according to cultivar and leaf age. Journal of Food Quality. 2019: 20.
36.    Kim GN, Jang HD. Flavonol content in the water extract of the mulberry (Morus alba L.) leaf and their antioxidant capacities. Journal of Food Science. 2011 Aug;76(6):C869-73.
37.    Li, J., Bai, Y., Zhang, P. and He, J., 2017. Simultaneous determination of 5 flavonoids and 7 saponins for quality control of traditional Chinese medicine preparation xinnaoshutong capsule using HPLC-VWD-ELSD. Journal of Analytical Methods in Chemistry. 2017: 3190185.
38.    International conference of harmonization Q2B, Validation of Analytical Procedures—Methodology, US FDA Federal Register, White Oak, MD, USA, 1997.
39.    JM B, PN B, MC R. Accuracy, precision, and reliability of chemical measurements in natural products research. Fitoterapia. 2011; 82: 44–52. doi:10.1016/J.FITOTE.2010.09.011.
40.    https://www.fda.gov/files/drugs/published/Analytical-Procedures-and-Methods-Validation-for-Drugs-and-Biologics.pdf.
41.    Ju WT, Kwon OC, Kim HB, Sung GB, Kim HW, Kim YS. Qualitative and quantitative analysis of flavonoids from 12 species of Korean mulberry leaves. J Food Sci Technol. 2018;55(5):1789-1796. doi:10.1007/s13197-018-3093-2.
42.    Polumackanycz M, Sledzinski T, Goyke E, et al. A Comparative Study on the Phenolic Composition and Biological Activities of Morus alba L. Commercial Samples. Molecules. 2019; 24: 3082. doi:10.3390/Molecules24173082.
43.    International Conference on Harmonisation of Technical requirements for registration of Pharmaceuticals for Human use ICH Harmonised Tripartite Guideline Validation of Analytical Procedures: Text and Methodology Q2(R1).

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