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目的:建立乳块消颗粒中极性和疏水性物质同时分析的LC-MS方法。方法:在LC-MS/MS系统下,使用高分子薄膜包被C10金刚烷基键合硅胶色谱柱(100 mm×2.1 mm,3μm),以0.1%甲酸水溶液为流动相A,乙腈为流动相B,梯度洗脱(0~4 min,0%B;4~14 min,0%B→90%B;14~16 min,90%B;16~16.1 min,90%B→0%B),流速0.2 m L·min~(-1),柱温35℃;采用电喷雾离子源(ESI),正离子多反应监测(MRM)模式进行含量测定。结果:在20 min内乳块消颗粒中9个成分实现良好分离,色谱峰面积精密度RSD<5%;标准曲线得到良好线性关系r2>0.997;样品回收率在97.2%~109.5%之间,RSD为3.8%~5.5%。所测6批样品中除尿嘧啶未检出外,尿苷、黄嘌呤、次黄嘌呤、刺桐碱、异荭草苷、牡荆素、芸香柚皮苷、橙皮素含量的依次为42.14~46.51、10.58~13.74、21.36~24.12、2.42~3.21、8.42~9.11、25.87~26.34、22.11~23.19、4.85~5.35μg·g~(-1)。结论:使用新型金刚烷基键合填料的反相色谱柱,可实现乳块消颗粒中的极性和疏水性成分同时保留;建立的LC-MS/MS法可同时测定乳块消颗粒中极性物质尿苷、尿嘧啶、黄嘌呤、次黄嘌呤、刺桐碱和疏水性物质异荭草苷、牡荆素、芸香柚皮苷、橙皮素9种化合物。经方法学验证,该方法简便,分离效率高,重复性好,可为乳块消颗粒的质量控制提供依据。
Objective: To establish a LC-MS method for the simultaneous analysis of polar and hydrophobic substances in Xiaobuxiao granules. METHODS: C10 adamantyl-bonded silica gel column (100 mm × 2.1 mm, 3 μm) was coated with polymer membrane under the LC-MS / MS system. The mobile phase consisted of 0.1% formic acid in water and acetonitrile B, gradient elution (0-4 min, 0% B; 4-14 min, 0% B → 90% B; 14-16 min, 90% B; 16-16.1 min, 90% B → 0% B) , The flow rate was 0.2 m L · min -1 and the column temperature was 35 ℃. Electrospray ionization (ESI) and positive ion multi-reaction monitoring (MRM) were used to determine the content. Results: Nineteen components of Niubixiao granule were separated well within 20 min, the RSD of chromatographic peak area was less than 5%, and the calibration curve showed a good linear relationship of r2> 0.997. The recoveries of samples were between 97.2% and 109.5% RSD is 3.8% ~ 5.5%. The six batches of samples tested except uracil were not detected, uridine, xanthine, hypoxanthine, tecorine, isoorientin, vitexin, rutin naringin, hesperetin content of 42.14 ~ 46.51,10.58 ~ 13.74,21.36 ~ 24.12,2.42 ~ 3.21,8.42 ~ 9.11,25.87 ~ 26.34,22.11 ~ 23.19,4.85 ~ 5.35μg · g -1. Conclusion: The reversed-phase column with the new adamantane-based packing material can be used for the simultaneous retention of polar and hydrophobic components in milk granules. The developed LC-MS / MS method can be used to determine the polar Urea, uracil, xanthine, hypoxanthine, pyriardine and hydrophobic substances isoorientin, vitexin, rutin naringin, hesperetin 9 kinds of compounds. The method is validated, the method is simple, high separation efficiency, good repeatability, which can provide the basis for the quality control of Xiaobu Xiaoguang.