RP-HPLC Method For The Determination Of Astragaloside Ⅳ, Glycyrrhizic Acid, Tanshinone Ⅱ A In Herbal Cosmetics

Nowadays,the development trend of cosmetics is to advocate green,environmental protection and pursuit the efficiency.In recent years,because of moderate healing effect of Chinese herbal medicines,Chinese herbal extracts as additives in cosmetics have become the hot spot.As we know,national departments concerned the cosmetics quality management is just only limited to the safe management,but never concerned on qualitative and quantitative detection of Chinese medicinal herbal components.Using derivatization reagent,astragalosideⅣand glycyrrhizic acid as the main ingredients of astragalus root and liquorice respectively were derived before chromatography,and new reversed phase high performance liquid charomatography methods for the determination of astragaloside IV,glycyrrhizic acid and tanshinoneⅡA in cosmetics were established for the first time.This paper mainly discussed derivatization reaction and chromatography analysis conditions. By one-factor analysis,the effect of amount of derivation reagent,the derivatization time,the derivatization reaction temperature and so on were investigated,moreover the effect of mobile phase,flow rate,column temperature,the detection wavelength and sampling volume were also researched indetail.Under the optimized derivatization reaction and chromatography analysis conditions,to the new methods,all of the parameters of three standard curves such as linear range,precision, detection limit and accuracy were researched one by one.The three new methods were used to the determination of the corresponding components in herbs and cosmetics extract liquids to examine the feasibilities of the methods.In addition,the reproducibilities of the methods were also checked by testing the real samples.Astragaloside IV was detected by reversed phase high-performance liquid chromatography (RPHPLC) using ultraviolet detection.By pre-column derivatization with 3,5-dinitrobenzoyl chloride,catalyzed by pyridine.The reaction mixtures were injected and separated on a C₁₈ column(5μm,4.6mm×150mm) with CH₃OH:KH₂PO₄(0.01mol·L^-1,adjust pH to 3.5 by H₃PO₄) =60:40(v/v) as mobile phase,and detected at 230 nm.Over the linear range of 5.0-100μg·mL^-1, the regression equation was y=478944+2132x,the correlation coefficient(R) was 0.9986,the precision was 1.56%(c= 40.0μg·mL^-1,n= 6),the detection limit was 1.2μg·mL^-1 and the average recovery was 98.6%,the new method of astragaloside IV was proved to be rapid,sensitive and feasible in real sample test.Using 4-nitrobenzyl bromide as derivative reagent,the procedure was carried out on a Phenomenex C₁₈(5μm,4.6mm×150mm) column.The mobile phase of CH₃CN:3%HAc=60:40 (v/v) was used throughout,and the flow rate was 1.0mL·min^-1.The experimental results were detected at 290nm.The new method for the determination of glycyrrhizic acid was over the linear range of 10.0-100.0μg·mL^-1 with the regression equation of y=369413+4032x,the correlation coefficient(R) was 0.9951,the precision was 1.67%(c= 10.0μg·mL^-1,n=6),the detection limit was 1.5μg·mL^-1 and the average recovery was 97.7%.In this paper,the analysis of tanshinoneⅡA was conducted by RPHPLC consisted of a C₁₈ column(5μm,4.6mm×150mm).With a column temperature of 30℃,an eluate of methanol:water (85:15) with a flow rate of 1.0mL·min^-1 and a detection wavelength at 270nm were used throughout.The standard curves were in the range of 1.0- 10.0μg·mL^-1 and 10.0-100.0μg·mL^-1 with the regression equations of y=280591+72704x and y=81572+97483x repectively,both of the two correlation coefficients were 0.9992.