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目的:采用高效液相色谱法同时测定复方卡力孜然凝胶剂中补骨脂素、异补骨脂素和蛇床子素的含量。方法:SHIMADZUVP-ODS色谱柱(4.6 mm×250 mm,5μm),柱温为35℃,以甲醇(A)和0.4%磷酸(B)为流动相,梯度洗脱[0～16 min,A-B(51:49),16～21 min,A-B(51:49)→A-B(74:26),21～35 min,A-B(74:26)],流速:1 mL·min~(-1),双波长检测:245nm(检测补骨脂素和异补骨脂素),322 nm(检测蛇床子素)。结果:样品中补骨脂素、异补骨脂素和蛇床子素分离良好,补骨脂素在2.31～23.10μg·mL~(-1)(r=0.9999)、异补骨脂素在1.92～19.20μg·mL~(-1)(r=0.9999)、蛇床子素在6.12～61.2μg·mL~(-1)(r=0.9998)范围内呈良好线性关系;补骨脂素、异补骨脂素和蛇床子素的回收率分别为97.2%(RSD为1.6%),96.3%(RSD为1.5%),97.9%(RSD为1.6%),样品溶液在12 h内稳定。结论:该方法简单准确,专属性强,重现性好,可作为复方卡力孜然凝胶剂中主要成分的理想检测方法。
Purpose: Simultaneous determination of psoralen, isopsoralen and osthole in compound calicheastrin gel by high performance liquid chromatography. METHODS: SHIMADZUVP-ODS column (4.6 mm×250 mm, 5 μm) was used. The column temperature was 35°C. Methanol (A) and 0.4% phosphoric acid (B) were used as mobile phase. Gradient elution [0～16 min, AB ( 51:49), 16-21 min, AB(51:49)→AB(74:26), 21-35 min, AB(74:26)], flow rate: 1 mL·min-1, double Wavelength detection: 245 nm (detects psoralen and isopsoralen), 322 nm (detects osthole). Results: Psoralen, isopsoralen, and osthole were well separated in the samples. Psoralen was 2.31-23.10 μg·mL -1 (r=0.9999) and isopsoralen was 1.92. There was a good linear relationship between ~19.20 μg·mL -1 (r=0.9999) and osthol in the range of 6.12 - 61.2 μg·mL -1 (r = 0.9998); psoralen, and supplement The recoveries of sulfatine and osthole were 97.2% (RSD 1.6%), 96.3% (RSD 1.5%), and 97.9% (RSD 1.6%), respectively, and the sample solution was stable within 12 h. Conclusion: The method is simple and accurate, with strong specificity and good reproducibility. It can be used as an ideal method for the detection of the main components in compound calichetran.