桔贝合剂HPLC特征图谱的建立和HPLC-ELSD同时测定6种成分含量

Establishment of Characteristic HPLC Chromatograms and Simultaneous Determination of Six Components by HPLC-ELSD of Jubei Mixture

关键词:  桔贝合剂  HPLC特征图谱  苦杏仁苷  甘草苷  贝母素甲  贝母素乙  桔梗皂苷D  麦冬皂苷D

Key words:  Jubei mixture  HPLC specific chromatogram  amygdalin  liquiritin  peimine  peiminine  platycodin D  ophiopogonin D

摘要:

目的 建立桔贝合剂的HPLC特征图谱,并采用HPLC-ELSD同时测定其中6种成分的含量。方法 建立桔贝合剂HPLC特征图谱,色谱柱为Agilent ZORBAX-C₁₈(250 mm×4.6 mm,5 μm),流动相为乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱,流速为1.0 mL·min^-1,柱温为30℃,检测波长为250 nm;含量测定采用HPLC-ELSD,色谱柱为Waters T3 C₁₈(250 mm×4.6 mm,5 μm),流动相为乙腈(A)-1%乙酸水溶液(B),梯度洗脱,流速为1.0 mL·min^-1,柱温为30℃。结果 建立了以黄芩苷为参照峰的HPLC特征图谱,确定了8个共有特征峰;苦杏仁苷、甘草苷、贝母素甲、贝母素乙、桔梗皂苷D和麦冬皂苷D在各自范围内线性关系良好(r ≥ 0.999),平均回收率分别为99.1%,98.6%,100.9%,97.7%,98.9%和98.0%,RSD分别为0.4%,1.0%,1.1%,1.2%,0.7%,1.3%。结论 本研究建立的方法可对桔贝合剂进行定性鉴别和定量测定,能够有效评价与控制桔贝合剂质量。

Abstract:

OBJECTIVE To establish characteristic HPLC chromatograms for Jubei mixture, and to simultaneous determine its six components by HPLC-ELSD. METHODS An HPLC specific chromatogram of the Jubei mixture was established. The chromatographic column was Agilent ZORBAX-C₁₈(250 mm×4.6 mm, 5 μm), gradient elution was conducted with acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) as the mobile phase, the flow rate was 1.0 mL·min^-1, and the column temperature was 30℃, the detection wavelength was 250 nm. An HPLC-ELSD method was established and the chromatography was accomplished on a Waters T3 C₁₈(250 mm×4.6 mm, 5 μm) column, the mobile phase consisted of acetonitrile(A)-1% acetic acid aqueous solution(B) in a gradient mode, the flow rate was 1.0 mL·min^-1, the column temperature was set at 30℃. RESULTS The characteristic chromatograms of the HPLC with baicalin as the reference peak was established, and eight common peaks were defined. The linear ranges of amygdalin, liquiritin, peimine, peiminine, platycodin D and ophiopogonin D were good within their own ranges(r ≥ 0.999). The average recovery were 99.1%, 98.6%, 100.9%, 97.7%, 98.9% and 98.0%, RSD were 0.4%, 1.0%, 1.1%, 1.2%, 0.7% and 1.3%, respectively. CONCLUSION The established methods can be used to qualitative identification and quantitative determination of Jubei mixture, can effectively used for evaluation and quality control of Jubei mixture.