Contents lists available at ScienceDirect Microchemical Journal journal homepage: www.elsevier.com/locate/microc A green approach for simultaneous analysis of two natural hepatoprotective drugs in pure forms, capsules and human plasma using HPLC-UV method Mohammed Gamal a,b, ⁎ , Hazim M. Ali c,d , Rehab M. Abdelfatah b , Maimana A. Magdy b a Pharmaceutical Chemistry Department, Faculty of Pharmacy, Jouf University, P.O. Box 2014, Sakaka, Aljouf, Saudi Arabia b Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Beni-Suef University, Alshaheed Shehata Ahmed Hegazy St., 62574 Beni-Suef, Egypt c Chemistry Department, College of Science, Jouf University, P.O. Box 2014, Sakaka, Aljouf, Saudi Arabia d Forensic Chemistry Department, Forensic Medicine Authority, Egypt ARTICLEINFO Keywords: Silymarin Vitamin E acetate HPLC Plasma Validation Greenness ABSTRACT A green high performance liquid chromatographic method with ultra violet detector (HPLC-UV) is presented, for the frst time, for the simultaneous estimation of two hepatoprotective drugs in capsules and spiked human plasma. The frst component is a herbal supplement, namely silymarin (SR) and the second is vitamin E acetate (VE). The method utilized an isocratic elution using C8 column, a mobile phase consisting of methanol:aceto- nitrile:triethylamine:trifuoroacetic acid (85:15:0.2:0.1, by volume) at 1 mL/min fow rate and 210 nm as a measuring wavelength. All silymarin favonolignans were measured together as a single peak. The method was evaluated for precision, selectivity, linearity, accuracy, LOQ, and LOD. SR and VE were detected in the linear range of 0.3–50 μg/mL and 0.1 to 50 μg/mL respectively. The Eco-scale score of the new HPLC method was 62 while it was 55 for the previous electrochemical method. Calibration curves had correlation coefcients of > 0.999. The new method is better than the old electrochemical voltammetric method in terms of simplicity, greenness, wide linear range and analysis time. 1. Introduction Herbal dietary supplements, such as silymarin (SR) and ginger, have signifcant variation in active constituents numbers and types. Many factors afect the herbal drug composition i.e. the quality of raw ma- terials, the extraction procedure, manufacturing and storage of the products. Therefore, sensitive methods should be established for eval- uating both starting materials and the fnal products [1]. Vitamin E acetate (VE) is a potent antioxidant whereas it reacts with the free ra- dicals and inhibits the oxidative damage of liver cell membranes [2]. Silymarin (SR) is a mixture of favonolignans and contains an anti- hepatotoxic polyphenolic molecules extracted from the milk thistle plant, Silybummarianum, family (Asteraceae), SR is composed of sili- binin, silicristin, and silidianin that have a free radical scavenging and hepatoprotective efects. Therefore, it has been applied for the treat- ment of various liver diseases that correlated with toxicity [1]. A cap- sule formulation of the two drugs is used for the treatment of decreased liver function as in the case of hepatitis; their chemical structures are shown in Fig. 1. After an extensive literature survey, several analytical techniques have been described for assaying SR in raw material and in its dosage forms including spectrophotometry [3–9], colorimetry [10], electro- chemical [11] and HPLC [5,12–20] methods, while VE was determined by colorimetric [21], spectrophotometry [22,23], gas chromatographic [24] and HPLC [25–27] methods. Walash et al. [5] separated SR successfully from dimethyl-4,4′-di- methoxy-5,6,5′,6′-dimethylene dioxybiphenyl-2,2′-dicarboxylate (DDB) in combined capsules, whereas a C18 column was utilized as a sta- tionary phase while the mobile phase was composed of 12% butanol, 0.17 M sodium dodecyl sulphate and 0.3% triethylamine, all solvents were prepared in 0.02 M phosphoric acid and the fnal pH was 3.5 in isocratic elusion system. The selected wavelength was 288 nm for UV detector and the linearity range was 1–15 μg/mL −1 for SR. Contrarily, the analysis of VE [25] was based on separation of the drug in dietary supplements on monolithic column Chromolith Per- formance RP-18 column while the mobile phase was consisted of me- thanol/water mixture (98:2, V/V) at a 2.0 mL/min fow rate. The de- tection wavelength was adjusted at 290 nm and the linearity range was 5–300 μg/mL −1 for VE. Only one method that is an electrochemical one [28] was found for the determination of both of SR and VE in combination, where the two drugs were determined simultaneously at a glassy carbon electrode in https://doi.org/10.1016/j.microc.2019.104258 Received 16 June 2019; Received in revised form 30 July 2019; Accepted 10 September 2019 ⁎ Corresponding author at: Pharmaceutical Chemistry Department, Faculty of Pharmacy, Jouf University, P.O. Box 2014, Sakaka, Aljouf, Saudi Arabia. E-mail address: mgamalm3000@yahoo.com (M. Gamal). Microchemical Journal 151 (2019) 104258 Available online 11 September 2019 0026-265X/ © 2019 Elsevier B.V. All rights reserved. T