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ABSTRACT

A sensitive, reproducible, and reliable reversed-phase high performance liquid chromatography (RP-HPLC) method with diode array detection (DAD) was developed and validated. Simultaneous determinations of five compounds; gallic acid, catechin, chlorogenic acid, epigallocatechin gallate and caffeic acid in four types of Momordica charantia extracts; water, ethanol, water:ethanol (1:1) and acetone were conducted. The compounds were successfully separated by C18 column (250 mm · 4.6 mm, 5 pm) with a gradient solvent system of 3% acetic acid in water:methanol:acetonitrile at flow rate of 1.0 mL/min. UV detection was carried out at 280 nm. The standard curves of the five compounds were linear in the range of 0.0396 pg/mL-100 pg/mL. The intra-assay relative standard deviation (RSD) was less than 4.97%, while the inter-assay RSD was less than 4.92%, whereas the accuracy was between 90.96% and 108.92%, respectively. Our optimised RP-HPLC-DAD method was capable to detect flavonoids and phenolic acid contents in four types of M. charantia fruits extracts simultaneously from five locations in Malaysia. The present method is recommended to be used for chemical analyses of phenolic compounds in other Momordica species.

Keywords: Momordica charantia, Phenolic compounds, Reversed-phase high performance liquid chromatography, Simultaneous determination

INTRODUCTION

Momordica charantia (M. charantia) L., known as bitter gourd and bitter melon, belongs to the Cucurbitaceae family (Habicht et al. 2011). It is a vegetable indigenous to the East and Southeast Asian countries, particularly Sri Lanka, Bhutan, Nepal, Philippines, Indonesia, Malaysia, Pakistan, Bangladesh, India, Taiwan, Thailand, China and Japan. Over the past decades, there is a large volume of published studies investigating the health and pharmacological activities of M. charantia. More than 100 studies (Basch, Gabardi and Bricht 2003; Grover and Yadav 2004; Lucas et al. 2010) have demonstrated that M. charantia has a strong association with broad health benefits: anticancer (Nagasawa, Watanabe and Inatomi 2002; Deep et al. 2004; Yasui et al. 2005, Dia and Krishnan 2016), anti-inflammatory, analgaesic (Lin and Tang 2008, Ullah et al. 2012), hypolipidemic and hypocholesterolaemic effects (Nerurkar, Lee and Nerurkar 2010). Moreover, extensive research has been explored on the effectiveness of fresh, juiced or dried M. charantia in diabetic animals and in type 2 diabetic human subjects (Welihinda and Karunanayake 1986; Grover and Yadav 2004; Dans et al. 2007, Mahmoud et al. 2017).

Phenolics are...

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