加味四妙颗粒的药效成分含量测定

目的：建立HPLC-DAD波长切换法同时测定加味四妙颗粒中延胡索乙素、盐酸小檗碱、盐酸黄柏碱、木兰花碱、杯苋甾酮、甘草苷、甘草次酸共7个成分的含量。方法：采用InsertSustain C₁₈色谱柱（250 mm×4.6 mm，5μm），以乙腈（A）-0.05 mol·L^-1磷酸二氢钾（B）为流动相，梯度洗脱，DAD检测器，检测波长268 nm（0~22.5 min，检测木兰花碱，67~71 min，检测盐酸小檗碱）、280 nm（22.5~30 min，检测延胡索乙素）、237 nm（30~45 min，检测甘草苷）、247 nm（45~55 min，检测杯苋甾酮，71~85 min，检测甘草次酸）、284 nm（55~67 min，检测盐酸黄柏碱）。结果：各待测组分分离度良好；延胡索乙素、盐酸小檗碱、盐酸黄柏碱、木兰花碱、杯苋甾酮、甘草苷、甘草次酸7个成分的进样质量浓度分别在2.060~51.49 μg·mL^-1（r=1.000）、3.111~77.77 μg·mL^-1（r=1.000）、1.813~45.31 μg·mL^-1（r=1.000）、1.960~49.00 μg·mL^-1（r=0.999 9）、2.019~50.48 μg·mL^-1（r=1.000）、2.468~61.70 μg·mL^-1（r=0.999 9）、2.408~60.20 μg·mL^-1（r=0.999 9）范围内与各自峰面积呈良好线性关系；平均加样回收率（n=6）分别为100.6%（RSD=1.6%）、100.3%（RSD=0.4%）、101.0%（RSD=1.3%）、100.7%（RSD=0.8%）、100.1%（RSD=1.9%）、101.6%（RSD=1.3%）、99.8%（RSD=1.7%）。结论：本试验建立的含量测定方法符合方法学验证要求，简便可靠，重现性好，可用于加味四妙颗粒的7个指标性成分的同时测定。

Objective:To establish a method for simultaneous determination of 7 active components(tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid)in Jiawei Simiao particles by HPLC-DAD method with wavelength switching. Methods:The separation was performed on a InsertSustain C₁₈(250 mm×4.6 mm,5 μm)column. The mobile phase was composed of acetonitrile(A)-0.05 mol·L^-1 potassium dihydrogen phosphate(B)with gradient elution. The detection wavelength was set at 268 nm(0-22.5 min,detection of magnoflorine,67-71 min,detection of berberine hydrochloride),280 nm(22.5-30 min,detection of tetrahydropalmatine)and 237 nm(30-45 min,detection of liquiritin,247 nm(45-55 min,detection of cyasterone,71-85 min,detection of glycyrrhetinic acid)and 284 nm(55-67 min,detection of phellodendrine chloride). Results:The ingredients could be effectively detected and separated. For tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid,the linear ranges were 2.060-51.49 μg·mL^-1(r=1.000),3.111-77.77 μg·mL^-1 (r=1.000),1.813-45.31 μg·mL^-1(r=1.000),1.960-49.00 μg·mL^-1(r=0.999 9),2.019-50.48 μg·mL^-1(r=1.000),2.468-61.70 μg·mL^-1(r=0.999 9)and 2.408-60.20 μg·mL^-1(r=0.999 9),respectively. The average recovery rate(n=6)were 100.6%(RSD=1.6%),100.3%(RSD=0.4%),101.0%(RSD=1.3%),100.7%(RSD=0.8%),100.1%(RSD=1.9%),101.6%(RSD=1.3%)and 99.8%(RSD=1.7%),respectively. Conclusion:The assay method established in this experiment met the requirements of methodological verification. The method was simple,reliable and reproducible. It could be used for simultaneous determination of 7 active components in Jiawei Simiao particles.