摘要
目的建立HPLC双波长指纹图谱快速测定藿香正气制剂中和厚朴酚含量的方法,建立糖量计法快速测定其总多糖含量的方法,并分析其差异性。方法 HPLC采用XB-C18(HS)色谱柱(4.6 mm×250 mm,5μm),乙腈和0.1%磷酸水溶液等度洗脱,流速1.0 mL/min,进样量20μL,检测波长294、270 nm,柱温30℃;糖量计法以50%乙醇为阴性对照;聚类分析重复性样品,对比不同指标间的聚类结果。结果 HPLC测得和厚朴酚在1.40~0.14 mg/mL范围内与峰面积线性关系良好,线性方程为A=12182633C-217418,r~2=0.9995;藿香正气水中和厚朴酚浓度为(0.4008±0.0050)mg/mL,藿香正气滴丸中其浓度为(0.4879±0.0089)mg/mL。总多糖在0.91~0.29 g/mL范围内与其百分值线性关系良好,线性方程为Y=51.949X+0.868,r~2=0.9999;藿香正气水中总多糖浓度为(0.31±0.003)g/mL,藿香正气滴丸中其浓度为(1.12±0.005)g/mL。聚类分析筛选出和厚朴酚峰面积和总多糖百分值作为表现藿香正气水和藿香正气滴丸间差异的指标。结论本研究建立的HPLC及糖量计法能有效用于藿香正气制剂多类指标的质量控制及不同剂型间的差异考察;聚类分析筛选出的峰面积和总多糖百分值作为评价指标具有代表性和准确性。
Objective To establish a method for rapid determination of honokiol in Huoxiang Zhengqi Preparations by HPLC dual-wavelength fingerprint, and to establish a sugar meter for rapid determination of total polysaccharide in Huoxiang Zhengqi Preparations, and to analyze its differences. Methods HPLC was performed on an XB-C18(HS) column(4.6 mm × 250 mm, 5 μm), with acetonitrile and 0.1% phosphoric acid aqueous solution for isocratic elution. The flow rate was 1.0 mL/min; the injection volume was 20 μL; the detection wavelength was 294 and 270 nm; the column temperature was 30 ℃. Sugar meter method was performed with 50% ethanol as negative control. Cluster analysis was conducted to analyze repeated samples and comparison of clustering results among different indexes. Results The linear relationship between the concentration of honokiol and the peak area in the range of 1.40–0.14 mg/mL was good by HPLC. The linear equation was A=12182633C-217418, r~2=0.9995. The concentration of honokiol in Huoxiang Zhengqi Liquid was(0.4008±0.0050)mg/mL; the concentration of honokiol in Huoxiang Zhengqi Dropping Pills was(0.4879±0.0089)mg/mL. The linear relationship between the concentration of total polysaccharide and its percentage was good in the range of 0.91–0.29 g/mL; the linear equation was Y=51.949X+0.868, r~2=0.9999. The concentration of total polysaccharide in Huoxiang Zhengqi Liquid was(0.31±0.003)g/mL; the total polysaccharide in Huoxiang Zhengqi Dropping Pills was(1.12±0.005)g/mL. The area of honokiol peak and the percentage of total polysaccharide as indicators of Huoxiang Zhengqi Liquid and Huoxiang Zhengqi Dropping Pills showed significant differences between different formulations. Conclusion Established HPLC and sugar meter method can be effectively used for the quality control and investigation of differences between different dosage forms of Huoxiang Zhengqi Preparations; the selected peak area and total polysaccharide percentage as the evaluation index are representative and accurate.
引文
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