HPLC法同时测定茵栀黄口服液中10种有效成分

Simultaneous determination of 10 active ingredients in Yinzhihuang Oral Liquid by HPLC

目的建立同时测定茵栀黄口服液中10种有效成分的HPLC方法,为茵栀黄制剂的质量控制、评价及标准修订提供科学依据。方法采用HPLC-UV法,色谱柱为DiamonsiL C18(250 mm×4.6 mm,5μm);流动相为乙腈-0.1%乙酸水溶液,梯度洗脱(0~15 min,8%~12%乙腈;15~40 min,12%~30%乙腈;40~55 min,30%~60%乙腈;55~75 min,60%~35%乙腈;75~80 min,35%~8%乙腈),体积流量1 mL/min,检测波长240 nm,柱温30℃,进样量20μL。结果同时测定了茵栀黄口服液中新绿原酸、绿原酸、隐绿原酸、栀子苷、对羟基苯乙酮、野黄芩苷、黄芩苷、槲皮素、黄芩素、汉黄芩素10种有效成分,各成分在考察的质量浓度范围内线性关系良好(r≥0.999 0),检测限与定量限分别为0.003~0.018μg/m L和0.009~0.055μg/m L,平均加样回收率为99.7%~102.6%,RSD为0.34%~6.14%。上述10种成分的平均质量浓度依次为(0.21±0.09)、(0.47±0.01)、(0.87±0.06)、(4.71±0.27)、(0.94±0.20)、(4.52±0.80)、(41.75±3.53)、(9.85±1.67)、(0.45±0.09)、(3.51±0.89)mg/mL,其中黄芩苷质量浓度为35.44~45.82 mg/m L,栀子苷质量浓度为4.16~4.92 mg/m L,均符合《中国药典》2015年版要求,质量合格。结论所建立的HPLC方法简单、专属、灵敏、稳定,精密度和准确度高,重现性好,可用于茵栀黄口服液质量控制和评价。

Objective To establish an HPLC method to simultaneously determine 10 active ingredients in Yinzhihuang Oral Liquid(YOL)and provide scientific basis for the quality control,evaluation and standard revision of Yinzhihuang preparations.Methods An HPLC-UV method was used with a Dikma Diamonsil C18 column(200 mm×4.6 mm,5μm).The mobile phase was acetonitrile-0.1%acetic acid solution with gradient elution(0—15 min,8%—12%acetonitrile;15—40 min,12%—30%acetonitrile;40—55 min,30%—60%acetonitrile;55—75 min,60%—35%acetonitrile;75—80 min,35%—8%acetonitrile).The detection wavelength was 240 nm.The flow rate was 1.0 mL/min.The column temperature was 30℃.The injection volume was 20μL.esults Ten active ingredients(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,geniposide,p-hydroxyphenylacetone,scutellarin,baicalin,quercetin,baicalein and wogonin)in YOL were simultaneously determined.The linearity was good(r≥0.999 0),the limit of detection and quantification were 0.003—0.018μg/m L and 0.009—0.055μg/mL.The average recoveries were 99.7%—102.6%with RSDs of0.34%—6.14%.The average content of the above 10 ingredients was in turn(0.21±0.09),(0.47±0.01),(0.87±0.06),(4.71±0.27),(0.94±0.20),(4.52±0.80),(41.75±3.53),(9.85±1.67),(0.45±0.09),(3.51±0.89)mg/mL.The content of baicalin and geniposide was 35.44—45.82 mg/mL and 4.16—4.92 mg/mL in eight batches,respectively.All eight batches of YOL meet the requirements of the Chinese Pharmacopoeia 2015 with qualified in quality.Conclusion The established HPLC method is simple,specific,sensitive,stable,precise,accurate,and reproducible,which can be used for quality control and evaluation of YOL.