UPLC-UV-MS法应用于胃肠安丸中11个活性成分的定性与定量分析

Quantitation and qualitation of eleven components in Weichang an pills by ultra-performance liquid chromatography-UV-tandem mass spectrometry

目的:建立超高效液相色谱-紫外-串联质谱法应用于胃肠安丸中11个活性成分(川芎嗪、阿魏酸、柚皮苷、橙皮苷、芦荟大黄素、大黄酸、大黄素、和厚朴酚、厚朴酚、大黄酚、大黄素甲醚)的定性、定量分析方法。方法:采用ACQUITY UPLCBEH C18(2.1 mm×50 mm,1.7μm)色谱柱,以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0～7.5 min,90%A→25%A),流速0.3 mL.min-1,DAD波长切换检测模式[0～1.92 min,320 nm(川芎嗪、阿魏酸);1.93～3.00 min,283 nm(柚皮苷、橙皮苷);3.01～5.50 min,254 nm(芦荟大黄素、大黄酸);5.51～6.26 min,294 nm(大黄素、和厚朴酚);6.27～7.50 min,254 nm(厚朴酚、大黄酚、大黄素甲醚)],柱温40℃,进样量2μL;Waters Quattro Premier XE质谱仪,正/负离子检测模式。结果:各成分在7.5 min内分离良好,通过与对照品的保留时间(tR)和质谱信息的对比,确定了胃肠安丸中的11个活性成分;它们在相应的浓度范围内与峰面积呈良好的线性关系,相关系数均>0.9977;加样回收率(n=5)为96.6%～107.2%,RSD均小于3.0%。批内精密度RSD均小于1.0%,批间精密度RSD均小于3.5%。结论:本方法简便、快速、准确,精密度高,重复性好,适用于胃肠安丸中多种活性成分的快速定性、定量分析。

Objective： To develop a simple, efficient and reliable method for simultaneous quantitation and qualita- tion of eleven compounds（ligustrazine, ferulic acid, naringin, hesperidin, aloe - emodin, rhein, emodin, honokiol, magnolol, ehrysophanol, physcion） in Weichang＇an pills by UPLC - UV - MS. Methods： The chromatographic separation was performed on a Waters ACQUITY UPLC system using an ACQUITY UPLC BEH C18 （2. 1 mm × 50 mm, 1.7 μm） column. The analysis was accomplished in 7.5 min with a gradient elution of water containing 0. 1% formic acid （A） and aeetonitrile （B） at flow rate of 0.3 mL.min^- 1 The switched wavelength of DAD was employed ： 0 - 1.92 min ： 320 nm （ ligustrazine, ferulic acid）, 1.93 - 3.00 min ： 283 nm （ naringin, hesperidin）, 3. 01 -5.50 min： 254 nm（aloe - emodin, rhein） 5.51 - 6.26 min ： 294 nm （ emodin, honokiol） 6. 27 -7.50min： 254 nm（ magnolol, chrysophanol, physcion）. The column temperature was set at 40 ℃. The injection vol- ume was 2μL. The mass spectra were obtained by Waters Quattro Premier XE mass spectrometer. The positive and negative ion modes were converted to detect the eleven compounds. Results：By comparing with the retention time （ tR） and mass spectrometry fragments data of reference substances, eleven compounds in Weichang＇an pill were i- dentified. All calibration curves were linear （ r 〉 0. 9977 ） over the tested ranges. The average recoveries ranged from 96.6% to 107.2% with RSD value below 3.0%. The RSDs of within - run and between - run precision were below 1.0% and 3.5%, respectively. Conclusions：The method is simple and fast with high precision, sensitivity and repeatability, which can be used for qualitative and quantitative analysis of multi - components in Weichang＇an pills.