乳块消颗粒中极性和疏水性成分的LC-MS/MS法同时测定

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，3 μm），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；16～16.1min，90% B→0% B），流速0.2 mL·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、4.85～5.35 μg·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 the method based on high performance liquid chromatography coupled to mass spectrometry(LC-MS/MS) for the simultaneous determination of hydrophilic and hydrophobic compounds in Rukuaixiao granules.Methods: The samples were separated on a CAPCELL PAK ADME-C10 column (100 mm×2.1 mm,3 μm) with 0.1% formic acid in water as mobile phase A and acetonitrile in water as mobile phase B using a gradient elution program (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 mL·min^-1,and the column temperature was 35℃. MS detection was performed with multiple reaction monitoring(MRM) using positive electrospray ionization. Results: The separation was completed within 20 min for the nine compounds,and the RSD of peak area precision was less than 5%.The standard curves obtained good linear relationships.The correlation coefficients of nine compounds were all higher than 0.997.The average recoveries (n=6) of the nine compounds were among 97.2%-109.5% with RSDs of 3.8%-5.5%.In the 6 batches of samples,uracil was not detected,the contents of the other eight ingredients were 42.14-46.51 μg·g^-1,10.58-13.74 μg·g^-1,21.36-24.12 μg·g^-1,2.42-3.21 μg·g^-1,8.42-9.11 μg·g^-1,25.87-26.34 μg·g^-1,22.11-23.19 μg·g^-1 and 4.85-5.35 μg·g^-1,respectively. Conclusion: The high new-style diamond alkyl bonded filler column can be used to obtain the simultaneous retention of the hydrophilic and hydrophobic compounds in Rukuaixiao granules and the established method based on LC-MS/MS can be applied for the simultaneous determination of uridine,uracil,2,6-dioxopurine,hypoxanthine, hypaphorine,homoorientin,vitexin,narirutin,and hesperetin.This method is proved to be simple with high separation efficiency and good reproducibility,and thus can provide reference for the quality control of Rukuaixiao granules.