目的:通过探讨补骨脂素苷与苷元酶解和酸水解的转化规律,建立补骨脂药材合理的质量评价方法。方法:采用Wonda Sil色谱柱(250 mm×4.6 mm,5 μm),以甲醇-0.4%磷酸水溶液(42∶58)为流动相,等度洗脱,流速1.0 mL·min-1,柱温35 ℃,紫外检测波长246 nm。结果:补骨脂苷、异补骨脂苷在酸水解中先产生中间体,并通过Q-TOF质谱检测器确认其结构,最后转化为补骨脂素、异补骨脂素,而通过β-葡萄糖苷酶则直接转化为补骨脂素、异补骨脂素。同时以酸水解提取方式建立含量测定方法,补骨脂素、异补骨脂素进样量分别在0.052~0.258 μg和0.123~0.616 μg范围内与峰面积线性关系良好(R2=1.000);平均回收率(n=6)分别为99.4%和100.5%,RSD分别为0.56%和0.83%,阴性对照无干扰。并用此方法测定了90批样品,补骨脂素和异补骨脂素的总和含量区间为1.69～2.64 mg·g-1。结论:通过酸水解的提取方式,更能科学地评价补骨脂中补骨脂素、异补骨脂素的真实含量,并可制定二者含量总和限度来控制补骨脂质量。
Objective: To establish a reasonable quality evaluation method for psoralen by discussing the transformation rule of enzymatic hydrolysis and acid hydrolysis of psoralen glycosides and aglycones. Methods: Wonda Sil chromatographic column (250 mm×4.6 mm,5 μm) was used. The mobile phase was methanol-0.4% phosphoric acid(42∶58) at a flow rate of 1.0 mL·min-1, the column temperature was 35 ℃, and the UV detector wavelength was 246 nm. Results: Psoralen and isopsoralen were first produced in acid hydrolysis, and their structures were confirmed by Q-TOF mass spectrometry detector. Finally, psoralen and isopsoralen were transformed into psoralen and isopsoralen directly by β-glucosidase. At the same time, the method of acid hydrolysis extraction was established to determine the content. Psoralen showed good linear rang with 0.052-0.258 μg (R2=1.000), the average recovery(n=6) was 99.4%,and RSD was 0.56%. Isopsoralen showed good linear in the range of 0.123-0.616 μg(R2=1.000), the average recovery(n=6) was 100.5%,and RSD was 0.83%. The negative control had no interference. 90 batches of samples were determined using the established method. The total contents of psoralen and isopsoralen ranged from 1.69 to 2.64 mg·g-1. Conclusion: The extraction method of acid hydrolysis can more scientifically evaluate the true content of psoralen and isopsoralen in Psoraleae Fructus, and can establish the sum limit of both contents to control the quality of Psoraleae Fructus.
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