ABSTRACT
OBJECTIVE：To establish the m ethod for simultaneous determination of 8 kinds of ginsenosides in Panax quinquefolium broken pieces. METHODS ：HPLC-DAD method was used to determine the contents of ginsenoside Rg 1，Re，Rb1， Rc，Ro，Rb2，Rb3，Rd in P. quinquefolium broken pieces. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid water solution （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Ginsenoside Re and ginsenoside Rb 2 were used as control ，liner calibration with two-reference substances correction was used to predict the retention time of other 6 components，and was compared with the relative retention time method. Using ginsenoside Re as control ， above components were quantified by the relative correction factor method ，and the results were compared with the external standard method. RESULTS ：The contents of ginsenoside Rg 1，Re，Rb1，Rc，Ro，Rb2，Rb3，Rd were 10.59-12.78，2.160-2.768， 27.492-38.880，3.154-4.018，3.368-4.080，0.343-0.755，0.961-1.415，5.857-6.923 mg/g. The accuracy of two-reference substances linear correction method to predict the retention time of components was higher ，and the absolute deviation of the predicted retention time was lower than that of the relative retention time method. There was no significant difference between the relative correction factor method and the external standard method ，and relative error was ＜3％ . CONCLUSIONS ：Established two-reference substances for determination of multiple components can be used for qualitative and quantitative analysis of 8 kinds of ginsensides in P. quinquefolium broken pieces simultaneously and accurately.
ABSTRACT
Substitute reference substance method is an effective approach for quality control of multiple components in accordance with the characteristics of traditional Chinese medicines. The purpose of the guideline is to guide the establishment of substitute reference substance method, prove the conformance of the method to the requirements for testing, and standardize the study method and its application in national drug standards. The topics of the guideline include the definition and classification of substitute reference substance method, the principles and approaches of quantitative analysis, the identification and confirmation of chromatographic peaks, and technical requirements. When substitute reference substance method is used for fingerprint identification or multiple components assay in traditional Chinese medicines, the analytical method can be validated following the guideline.