"Swissica Brand" American Ginseng Royal Jelly Oral Liquid is made from American Ginseng, Schisandra chinensis, Fresh Royal Jelly and Honey. Ginsenoside is the main component of American Ginseng, 10-hydroxy-2-decenoic acid (hereinafter referred to as Decenoic Acid) is the active ingredient of royal jelly The quality of the preparation can be evaluated by measuring the ginsenoside and decenoic acid in the oral solution. The internal control method is to extract the samples by different methods, determine the total saponin by colorimetry, and determine the decenoic acid by thin layer scanning The method is time-consuming and requires a large amount of solvent. The method for determining saponins by HPLC has been reported in literature [1 ~ 6], but there is no report for the simultaneous determination of ginsenosides and decenoic acid. In this paper, a Sep-pak small column is used to process samples After that, the method of simultaneous determination of ginsenoside and decenoic acid by HPLC is simple, accurate and time-saving.
1 Experimental part
1.1 Instruments and reagents STI5000 liquid chromatograph, UV5000 detector, N2000 chromatography workstation.
Acetonitrile is an HPLC reagent. The remaining reagents are of analytical grade, and the water is double distilled water.
Ginsenosides (Re, Rg1, Rb1), decenoic acid reference substance (China National Institute for the Control of Pharmaceutical and Biological Products). American Ginseng Royal Jelly Oral Liquid Sample (Experimental Pharmaceutical Factory, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences).
1.2 Chromatographic conditions Chromatographic column YWGC18 stainless steel column (250mm × 4.6 mm, 5μm), Waters C18 guard column; Varian chromatography workstation. Mobile phase: acetonitrile-methanol-water (30: 5: 70) for determination of Re, Rg1 and decenoic acid; acetonitrile-water (40:60) for determination of Rb1. Flow rate 1ml. min-1; detection wavelength 203nm. Under the above chromatographic conditions, the retention time of Re + Rg1 and decenoic acid is about 8-9min and 11min respectively; the retention time of Rb1 is about 5.5min, the chromatograms are shown in Figure 1 and Figure 2.
Figure 1 HPLC chromatogram of Re + Rg1 and decenoic acid
A. Standard B. Sample 1. Re + Rg1 2. Decenoic acid 3. Internal standard
Figure 2 HPLC chromatogram of Rb1
A. Standard B. Sample 1. Rb1
1.3 Investigation of the linear relationship Precisely weigh about 10 mg each of ginsenoside Re, Rg1 and decenoic acid in a 10 ml volumetric flask, make up to volume with methanol to prepare a mixed standard stock solution, and then dilute this solution in a concentration range of 10-100 μg. ml-1. The Rb1 standard series is prepared by diluting the Rb1 standard (1mg.ml-1) in a concentration range of 10 to 100μg. ml-1. Inject 20μl of the standard series solution in 10 ～ 100μg. ml-1, get the regression equation: Re + Rg1 is Y = -14363 + 1793.1X, r = 0.9995; decenoic acid is Y = 1694 + 25883X, r = 0.9999; Rb1 is Y = -1289.7 + 1801.4X, r = 0.9999.
1.4 Precision and detection limit Accurately prepare each reference solution equivalent to the concentration of the sample solution, and measure it 5 times within a day to get the intra-day precision (RSD%): Re + Rg1 is 2.9%, and decenoic acid is 1.3
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