HPLC-MS/MS法测定桑菊感冒片中5个成分的含量

目的：建立高效液相色谱-串联质谱法同时测定桑菊感冒片中芦丁、绿原酸、连翘苷、桔梗皂苷D和苦杏仁苷的含量。方法：采用Atlantis C₁₈色谱柱（2.1 mm×150 mm，5 μm），以乙腈（A）-0.1%甲酸（B）为流动相，梯度洗脱（0~3 min，20% A；3~15 min，20% A→60% A；15~23 min，60% A；23~24 min，60% A→20% A；24~30 min，20% A），流速0.2 mL·min^-1，柱温35℃，采用电喷雾离子源（ESI），以多反应监测（MRM）方式进行正离子扫描。结果：芦丁、绿原酸、连翘苷和桔梗苷D质量浓度均在0.01~2 μg·mL^-1的范围内线性关系良好（r>0.999），苦杏仁苷质量浓度在0.25~50 μg·mL^-1的范围内线性关系良好（r>0.999），平均回收率（n=6）分别为98.9%、99.8%、100.6%、99.4%、99.6%，RSD分别为0.84%、0.90%、1.2%、1.3%、0.86%；3批样品中芦丁、绿原酸、连翘苷、桔梗皂苷D和苦杏仁苷含量范围分别为244.0~267.9、257.3~278.8、170.9~190.0、169.3~177.8、4763.9~4872.5 μg·g^-1。结论：所建立的方法为桑菊感冒片的全面质量控制提供科学依据。

Objective:To establish a method for the determination of rutin,chlorogenic acid,forsythin,platycodin D and amygdalin in Sangju Ganmao tablets by HPLC-MS/MS.Methods:The determination was carried out on an Atlantis C₁₈ column(2.1 mm×150 mm,5 μm)at 35℃.The mobile phase consisted of acetonitrile(A)-0.1% formic acid solution(B)with gradient elution(0-3 min,20%A;3-15 min,20%A→60%A;15-23 min,60%A;23-24 min,60%A→20%A;24-30 min,20%A)at a flow rate of 0.2 mL·min^-1.The analytes were detected by tandem mass spectrometry with the electrospray ionization(ESI)source combined with multiple reaction monitoring(MRM)modes.Results:Rutin,chlorogenic acid,forsythin and platycodin D showed good linearities in the range of 0.01-2 μg·mL^-1(r>0.999).Amygdalin showed a good linearity in the range of 0.25- 50 μg·mL^-1(r>0.999).The average recoveries(n=6)were 98.9%,99.8%,100.6%,99.4% and 99.6% with RSDs of 0.84%,0.90%,1.2%,1.3% and 0.86%.The contents of rutin,chlorogenic acid,forsythin,platycodon D and amygdalin in 3 batches of samples were 244.0-267.9 μg·g^-1,257.3-278.8 μg·g^-1,170.9-190.0 μg·g^-1,169.3-177.8 μg·g^-1 and 4 763.9-4 872.5 μg·g^-1,respectively.Conclusion:This method can be used for the overall assessment of the quality of Sangju Ganmao tablets.