基于HPLC-CAD的元胡止痛口服液糖类成分定量指纹图谱研究

Development of a Quantitative Chromatographic Fingerprint Analysis Method for Sugar Components of Yuanhu Zhitong Koufuye Using HPLC-CAD

关键词:  HPLC-CAD  元胡止痛口服液  糖类  定量指纹图谱

Key words:  HPLC-CAD  Yuanhu Zhitong Koufuye  sugar  quantitative chromatographic fingerprint

摘要:

目的 采用高效液相色谱-电喷雾检测器(HPLC-CAD)检测技术建立元胡止痛口服液中糖类成分的定量指纹图谱分析方法。方法 采用NH₂P-50 4E色谱柱(4.6×250 mm,5 μm),以水(A)-乙腈(B)为流动相进行梯度洗脱,柱温为30 ℃,进样量为10 μL,流量为0.6 mL/min,CAD雾化温度为35 ℃,采样频率10 Hz,幂函数1.0。结果 定量成分在定量范围内线性关系良好,R2＞0.999,进样精密度、中间精密度、方法重复性的RSD值均小于3 %,供试品溶液在24 h内稳定,低、中、高三个水平浓度的平均加样回收率为97.15 %~101.13 %。指纹图谱中含共有峰5个,共有峰面积占总峰面积比例为93.77%,其精密度、重复性、稳定性的RSD值均小于4%。结论 建立的分析方法稳定、准确、重复性好,能够检测制剂中的糖类辅料。

Abstract:

OBJECTIVE A quantitative fingerprint analysis method was established for sugar components in Yuanhu Zhitong Koufuye using high performance liquid chromatography (HPLC) -charged aerosol detection (CAD). METHODS Chromatographic column was NH₂P-50 4E column (4.6×250 mm, 5 μm). Water (A) and acetonitrile (B) were used as the mobile phase in the gradient elute mode. The column temperature was 30 ℃. The injection volume was 10 μL. The flow rate was 0.6 mL/min. The evaporation temperature of CAD was 35 ℃. The acquisition frequency was 10 Hz. The power function value was 1.0. RESULTS The linear relationship of the quantitative component was good within the quantitative range. The value of R2 > 0.999. The relative standard deviations (RSDs) of injection precision, intermediate precision and method repeatability were all less than 3 %. The test solution was stable within 24 h. The average recoveries at three levels ranged was 97.15 %~101.13 %. There were 5 common peaks in the fingerprint. The proportion of common peak area to total peak area was 93.77 %. The RSDs of injection precision, method repeatability and sample stability were all less than 4%. CONCLUSION The established analytical method is stable, accurate and reproducible. It can be used to detect sugar excipients in the preparations.