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Simultaneous determination of six active components in crude and processed Fructus Corni by high performance liquid chromatography

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Abstract

A simple and rapid HPLC method was established for simultaneously determining six active components in Fructus Corni. The six components were separated on an Agilent Zorbax Extend C18 column (250 mm × 4.6 mm, 5 μm) and detected by diode array detector (DAD). Mobile phase was composed of (A) aqueous phosphoric acid (0.1%, v/v) and (B) acetonitrile phosphoric acid (0.1%, v/v) using a gradient elution. Analyses were performed at 30 °C with a flow rate of 1.0 mL/min and UV detection at 218 nm, 240 nm and 284 nm. All calibration curves showed good linear regression (r2  0.9999) within tested ranges. The LOD and LOQ were 0.11–1.69 μg/mL and 1.48–16.60 μg/mL, respectively. Overall intra-day and inter-day variations were less than 4.72%, and the average recoveries were 97.97–102.51% for the analytes. The developed method can be applied to the intrinsic quality control of Fructus Corni.

Introduction

Fructus Corni is derived from the dry ripe sarcocarp of Cornus officinalis Sieb. et Zucc. The crude drug or its processed products of jiu zheng pin (JZP) and qing zheng pin (QZP) are used clinically. JZP is produced after steaming the crude drug pre-steeped in wine, and QZP is produced after steaming the crude drug [1], [2].

In the last decades, Fructus Corni has been extensively investigated in phytochemistry, and the results indicated that gallic acid, 5-hydroxymethylfurfural (5-HMF), morroniside, sweroside, loganin and cornuside were the main active components in Fructus Corni. Pharmacological studies on the components showed that they all had good biological activities. Gallic acid had anti-inflammatory activity [3] and bacteriostatic action [4], [5]. Morroniside and loganin showed antioxidation and protective effects on rat mesangial cell proliferation [6]. Sweroside could resist d-aminogalactose hepatic injury [7]. Cornuside suppressed cytokine-induced proinflammatory, relaxated vasculature, and inhibited melanogenesis [8], [9], [10]. 5-HMF could improve blood circulation [11], [12].

To our knowledge, previously reported analytical methods were developed to quantify only one or two types of components in Fructus Corni such as gallic acid [13], [14], [15], morroniside [16] and loganin [17], [18], [19]. In current study, an HPLC-DAD method was developed to quantify six components simultaneously in Fructus Corni.

Section snippets

Samples, chemicals and reagents

Fructus Corni was collected from four suppliers (Henan, Shanxi, Zhejiang and Anhui in China). Gallic acid was purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). 5-HMF was purchased from Shanghai Yousi Biotechnology Co. Ltd. (Shanghai, China). Morroniside, sweroside and loganin were purchased from Institute of Jiangsu Pharmaceutical Research (Nanjing, China). Cornuside was prepared from our laboratory, and its identity was verified

Optimization of chromatographic conditions

Different mobile phase compositions were tested: (1) water–methanol; (2) water–acetonitrile; (3) aqueous phosphoric acid (0.1%, v/v)–acetonitrile phosphoric acid (0.1%, v/v); (4) aqueous ammonium acetate (0.5%, v/v)–acetonitrile. As a result, the combination of aqueous phosphoric acid (0.1%, v/v)–acetonitrile phosphoric acid (0.1%, v/v) for mobile phase was the best for separation. Furthermore, other chromatographic variables were also optimized, including analytical columns (Hanbon Hedera

Conclusion

An HPLC-DAD method has been developed to simultaneously determine six active components in Fructus Corni. This newly established method is validated as simple, precise and accurate. It can be used as a valid analytical method for intrinsic quality control of Fructus Corni.

Acknowledgement

This research was financially supported by the fund of national science and technology research item in “10th Five Year Plan” (2001BA701A11).

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    These authors contribute equally to this work.

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