HPLC法同时测定清肺抑火片中14种成分的含量

目的 建立高效液相色谱法(HPLC)同时测定清肺抑火片中栀子苷、芒果苷、黄芩苷、盐酸小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、白花前胡甲素、大黄酚、大黄素甲醚、白花前胡乙素14种成分含量的方法。方法 采用Titank C₁₈ (250 mm×4.6 mm,5 μm)色谱柱；以乙腈-0.1%磷酸溶液为流动相,梯度洗脱；检测波长: 黄芩苷、汉黄芩苷、黄芩素、汉黄芩素为280 nm,芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、芒果苷为254 nm,栀子苷为238 nm,盐酸小檗碱为265 nm,白花前胡甲素、白花前胡乙素为321 nm；流速为1.0 mL? min-1； 柱温为35 ℃； 进样量为10 μL。结果 栀子苷、芒果苷、黄芩苷、盐酸小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、白花前胡甲素、大黄酚、大黄素甲醚、白花前胡乙素的线性范围分别为:4.96～223.17、0.84～42.22、18.76～938.16、4.46～223.17、4.86～243.10、1.59～79.32、0.76～38.17、1.03～51.49、1.59～79.40、1.21～60.72、1.80～90.06、0.91～45.48、1.04～51.83、0.86～43.23μg?mL?1,r均≥ 0.9999。精密度、稳定性、重复性试验的RSD均＜2%,平均加样回收率(n=6)均＞90%,RSD＜3%。结论 该方法操作简单,重现性好,可为完善清肺抑火片的质量标准提供科学依据。

Simultaneous Determination of 14 Components in Qingfei Yihuo Tablets by HPLC

ABSTRACT: OBJECTIVE To establish a high-performance liquid chromatography method for the simultaneous determination of 14 components including geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, praeruptorinA, chrysophanol, physcion and praeruptorin B in Qingfei Yihuo tablets. METHODS Titank C18 (250 mm×4.6 mm, 5 μm) column was used; acetonitrile-0.1% phosphoric acid solution was used as the mobile phase with gradient elution; detection wavelengths: baicalin, wogonoside, baicalein, and wogonin at 280 nm, aloe-emodin, rhein, emodin, chrysophanol, physcion and, mangiferin at 254 nm, geniposide at 238 nm, berberine hydrochloride at 265 nm, praeruptorinA and praeruptorinB at 321 nm; The flow rate was 1.0 mL?min-1 and the column temperature was 35℃; the injection volume was 10 μL. RESULTS The linear ranges of geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein,aloe-emodin, rhein, wogonin, emodin, praeruptorinA, chrysophanol, physcion and, praeruptorin B were: 4.96~223.17, 0.84~42.22, 18.76~938.16, and 4.46~223.17, 4.86~243.10, 1.59~79.32, 0.76~38.17, 1.03~51.49, 1.59~79.40, 1.21~60.72, 1.80~90.06, 0.91~45.48, 1.04~51.83, 0.86~43.23 μg?mL?1, all with r ≥ 0.9999. The RSDs of precision, stability and reproducibility tests were <3%, and the average recoveries in sample (n=6) were >90%, with the RSDs was <3%. CONCLUSION The method is simple and reproducible and can provide a scientific basis for improving the quality standard of Qingfei Yihuo Tablets.