采用LC-ESI/MS方法同时测定荆防败毒口服液中4个有效成分含量

目的：建立同时测定荆防败毒口服液中4个有效成分（升麻素苷、紫花前胡苷、二氢欧山芹醇当归酸酯和欧当归内酯A）含量的LC-ESI/MS方法。方法：通过优化，建立了如下方法同时测定荆防败毒口服液中4个有效成分（升麻素苷、紫花前胡苷、二氢欧山芹醇当归酸酯和欧当归内酯A）：采用ACQUITY UPLCBEH C₁₈色谱柱（2.1 mm×50 mm，1.7 μm），柱温为30℃，以0.1%甲酸水溶液（A）-乙腈（B）为流动相，梯度洗脱（0~10 min，90% A → 0% A；10~11 min，0% A → 90% A；11~13 min，90% A），流速0.2 mL·min^-1，分析时间为13 min，进样量1 μL；采用ESI正负离子模式，多反应监测（MRM），应用Masslynx V4.1软件进行数据分析。结果：升麻素苷、紫花前胡苷、二氢欧山芹醇当归酸酯和欧当归内酯A能够在13 min内完全分离，质量浓度在一定范围内与峰面积呈良好的线性关系（r²>0.998），日内精密度RSD为0.19%~0.68%，日间精密度RSD为0.74%~1.9%，重复性RSD为0.11%~1.9%，稳定性RSD为3.3%~8.6%，加样回收率在97.7%~102.3%（RSD为0.50%~2.7%）；测得的4批样品中升麻素苷、二氢欧山芹醇当归酸酯、紫花前胡苷和欧当归内酯A的含量范围分别为2.27~2.52、6.35~7.06、143.14~180.534和0.076~0.186 μg·mL^-1。结论：该方法可以用于荆防败毒口服液中4个有效成分的含量测定和质量控制。

Objective: To establish a LC-ESI/MS method for simultaneous determination of four bioactive constituents (prim-O-glucosylcimifugin, nodakenin,columbianadin and levistolide A)in Jingfangbaidu oral liquid.Methods: The separation was performed on an ACQUITY UPLC BEH C₁₈ column(2. 1 mm×50 mm,1. 7 μm)with gradient elution(0-10 min, 90%A → 0%A; 10-11 min, 0%A → 90%A; 11-13 min, 90%A)using 0. 1% formic acid (A)and acetonitrile(B)as mobile phase. And the column temperature,flow rate,injection volume and analysis time were 30℃,0. 2 mL·min^-1, 0. 1 μL and 13 min, respectively. Quantification of the analytes was performed on a Triple quadrupole mass spectrometer. A multiple-reaction monitoring scanning and an electrospray ionization(ESI)-tandem interface in the positive and negative ion polarity mode were employed. Data analysis was operated on Masslynx V4. 1 software.Results: Four ingredients,prim-O-glucosylcimifugin,nodakenin,columbianadin and levistolide A could be separated in 13 min,and all calibration curves showed good linearity(r²>0. 998)within the test range. The RSD of intra-day precision,inter-day precision,repeatability and stability were within 0. 19%-0. 68%,0. 74%-1. 9%, 0. 11%-1. 9% and 3. 3%-8. 6%,respectively. The average recoveries for the four analytes were between 97. 7% and 102. 3%,and the RSD was within 0. 50%-2. 7%. The contents of prim-O-glucosylcimifugin,nodakenin,columbianadin and levistolide A in four batches of samples were within 2. 27-2. 52 μg·mL^-1, 6. 35-7. 06 μg·mL^-1, 143. 14-180. 534 μg·mL^-1 and 0. 076-0. 186 μg·mL^-1,respectively.Conclusion: The proposed method could be used to determine the four components and quality control of Jingfangbaidu oral liquid.