基于UPLC-TQ/MS联用技术分析乳香-没药不同比例配伍化学成分溶出变化

目的：建立同时测定乳香、没药中16个化学成分的UPLC-TQ/MS方法，对乳香-没药7个不同配伍比例的化学成分溶出变化进行定量分析；从化学成分角度分析乳香、没药不同配伍比例中的化学成分溶出变化，以期确定最优配伍比例，为进一步阐明量效关系和指导临床用药提供依据。方法：应用UPLC-TQ/MS联用技术对乳香-没药不同配伍比例（1∶1，2∶1，1∶2，2∶3，3∶2，3∶4，5∶3）的化学成分溶出变化进行分析。采用Acquity^TMUPLC BEH C₁₈色谱柱（2.1 mm×50 mm，1.7 μm），以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱，流速0.3 mL·min^-1，采用UPLC-TQ/MS的电喷雾正、负离子源（ESI⁺/ESI^-），多反应检测（MRM）模式。结果：乳香-没药为2∶1配伍的16个化学成分总含量最高（1.664%）且浸膏得率较高（59.56%）；16个化合物中，3α-乙酰基-甘燧-7，24-二烯-21-酸（0.146%）、乙酰11α-甲氧基-β-乳香酸（0.079%）、β-乳香酸（0.210%）、3β-乙酰氧基-5α-8，24-羊毛脂二烯-21-酸（0.058%）、3α-乙酰氧基-羊毛脂-8，24-二烯-21-酸（0.197%）在乳香-没药比例为1∶1配伍时含量最高；3-羟基甘遂烷-8，24-二烯-21-酸（0.164%）、3-乙酰基氧基甘遂-8，24-二烯-21-羧酸（0.065%）、松香酸（0.017%）、3-α羟基甘燧烷-7，24-二烯-21-酸（0.140%）在乳香-没药比例为2∶1配伍时含量最高；3-O-乙酰基-α乳香酸（0.189%）在乳香-没药比例为3∶2配伍时含量最高；2-甲氧基-8，12-环氧吉马烷-1（10），7，11-三烯-6-酮（0.265%）、3-乙酰基-11-酮基-β-乳香酸（0.267%）、2-甲氧基-5-乙酰基-呋喃吉马烷-1（10）-烯-6-酮（0.058%）在乳香-没药比例为3∶4配伍时含量最高；α-乳香酸（0.149%）、11-酮基乳香酸（0.036%）在乳香-没药比例为5∶3配伍时含量最高。其中，乙酰11α-甲氧基-β-乳香酸仅在配伍比例为1∶1配伍时检测到，16个化合物在乳香-没药1∶1配伍时全部检测出。结论：乳香-没药不同配伍比例的化学成分溶出不同，且配伍比例为1∶1和2∶1时溶出化学成分种类最为丰富，成分含量最高或较高，且该2种配伍比例也是临床用药的常用比例，该研究结果为进一步阐明乳香-没药配伍的量效关系和指导临床用药提供了科学依据。

Objective: To establish an UPLC-TQ-MS method for simultaneous determination of 16 components in frankincense-myrrha compatibility for quantitative analysis of dissolution changes of chemical components in 7 different compatibility ratios of frankincense and myrrha,with an expectation to determine the optimal compatibility ratio for further clarification of the dose-effect relationship and guidance for clinical medication. Methods: Dissolution changes of chemical components in different compatibility ratios of frankincense and myrrha (1:1,2:1,1:2,2:3,3:2,3:4 and 5:3) were determined by UPLC-TQ/MS. The separation was performed on an Acquity TM UPLC BEH C₁₈ (2.1 mm×50 mm,1.7 μm) column. And the mobile phase consisted of 0.1% formic acid-acetonitrile with gradient elution at a flow rate of 0.3 mL·min^-1. Both the positive and negative scanning modes were applied in the multiple reaction monitoring (MRM). Results: The total contents of 16 chemical components (1.664%) and the extract yield (59.56%) were highest in frankincense-myrrha with 2:1 compatibility. Among 16 compounds,3α-acetoxy-tirucall-7,24-dien-21-oic acid (0.146%),acetyl 11α methoxy-β-boswellic acid (0.079%),β-boswellic acid (0.210%),3β-acetoxy-5α-lanosta-8,24-dien-21-oic acid (0.058%) and 3α-acetyloxy-lanosta-8,24-dien-21-oic acid (0.197%) were most abundant in frankincense-myrrha with 1:1 compatibility. 3-hydroxytirucall-8,24-dien-21-oic acid (0.164%),3-acetyloxy-tirucall-8,24-dien-21-oic acid (0.065%),abietic acid (0.017%)and 3α-hydroxy tirucall-7,24-dien-21-oic acid (0.140%) were most abundant in frankincense-myrrha with 2:1 compatibility. 3-O-acetyl-α-boswellic acid (0.189%) was most abundant in frankincense-myrrha with 3:2 compatibility. 2-methoxy-8,12-epoxygermacra-1 (10),7,11-trien-6-one (0.265%),3-acetyl-11-keto-β-boswellic acid (0.267%) and 2-methoxy-5-acetoxy-fruranogermacr-1 (10)-en-6-one (0.058%) were most abundant in frankincense-myrrha with 3:4 compatibility. α-boswellic acid (0.149%) and 11-keto-boswellic acid (0.036%) were most abundant in frankincense-myrrha with 5:3 compatibility. Acetyl 11α-methoxy-β-boswellic acid was detected only in frankincense-myrrha with 1:1 compatibility,and 16 compounds were all detected in the above compatibility. Conclusion: The dissolutions of chemical components in the different ratios of frankincense and myrrha compatibility are different,and most richest or most abundant soluble chemical components are found when the compatibility ratios are 1:1 and 2:1,which are also the commonly used in clinical medicine. The study provides a scientific basis for further elucidating the dose-effect relationship of frankincense-myrrha compatibility and guiding clinical medication.

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