摘要
目的建立同时测定肋柱花药材中5种活性成分含量的超高效液相色谱-三重四极杆串联质谱法。方法色谱柱为Thermo Hypersil GOLD C_(18)柱(100 mm×2.1 mm,1.9μm),流动相为甲醇-含5 mmol/L甲酸铵的0.1%甲酸水溶液(梯度洗脱),流速为0.25 mL/min,柱温为30℃,进样量为5μL;离子源为电喷雾电离(ESI),电离模式为ESI^(-)(木犀草素、芒果苷)和ESI^(+)(异荭草苷、獐牙菜苦苷、当药黄素),多反应监测(MRM)模式,喷雾电压为3500 V(正离子模式)和3000 V(负离子模式),雾化温度为300℃。采用SIMCA 14.1软件进行主成分分析。结果木犀草素、异荭草苷、獐牙菜苦苷、芒果苷、当药黄素的质量浓度分别在0.4041~12.9312 mg/L、0.3922~12.5504 mg/L、0.2838~9.0816 mg/L、0.3055~9.7760 mg/L、0.3567~11.4144 mg/L范围内与峰面积线性关系良好(R2≥0.9990);检测限分别为0.5210,1.3198,0.8133,1.2194,2.0830μg/L,定量限分别为1.5109,3.9312,2.7096,4.0241,6.7365μg/L;精密度、稳定性、重复性试验结果的RSD均小于3.5%(n=6);加样回收率分别为94.83%,92.30%,100.47%,95.13%,95.53%,RSD分别为1.49%,2.50%,0.92%,0.91%,1.48%(n=6)。12批不同产地的药材样品中5种活性成分的总含量为39.85~67.82 mg/g,獐牙菜苦苷的含量最多(33.87~61.44 mg/g)。主成分分析结果显示,第一主成分、第二主成分的方差贡献率分别为36.30%和31.70%,筛选出的主要贡献成分为木犀草素(0.6230)、獐牙菜苦苷(0.5205)和芒果苷(0.7533)。结论该方法操作简便、专属性强、结果准确、灵敏度高,可用于肋柱花药材中活性成分的定量分析,木犀草素、獐牙菜苦苷、芒果苷可作为肋柱花药材的质量控制标志物。
Objective To establish an ultra-high performance liquid chromatography triple-quadrupole-tandem mass spectrometry(UPLC-QQQ-MS/MS)method for the simultaneous determination of five active components in Lomatogonii Rotati Herba.Methods The chromatographic column was Thermo Hypersil GOLD C_(18)column(100 mm×2.1 mm,1.9µm),the mobile phase was methanol-0.1%formic acid aqueous solution containing 5 mmol/L ammonium formate(gradient elution),the flow rate was 0.25 mL/min,the column temperature was 30℃,and the injection volume was 5µL.The ion source was electric spray ionization(ESI),the ionization mode was ESI^(-)(luteolin,mangiferin)and ESI^(+)(isoorientin,swertimarin and swertisin),the multi-reaction monitoring(MRM)mode was adopted with spray voltages of 3500 V(positive ion mode)and 3000 V(negative ion mode),and the atomization temperature was 300℃.SIMCA 14.1 software was used for principal component analysis(PCA).Results The linear ranges of luteolin,isoorientin,swertimarin,mangiferin and swertisin were 0.4041-12.9312 mg/L,0.3922-12.5504 mg/L,0.2838-9.0816 mg/L,0.3055-9.7760 mg/L,0.3567-11.4144 mg/L(R2≥0.9990),respectively.The limits of detection(LOD)were 0.5210,1.3198,0.8133,1.2194 and 2.0830µg/L,respectively.The limits of quantitation(LOQ)were 1.5109,3.9312,2.7096,4.0241 and 6.7365µg/L,respectively.The RSDs of precision,stability,and repeatability test results were all lower than 3.5%(n=6).The average recoveries of luteolin,isoorientin,swertimarin,mangiferin and swertisin were 94.83%,92.30%,100.47%,95.13%,and 95.53%,with RSDs of 1.49%,2.50%,0.92%,0.91%,and 1.48%(n=6),respectively.The total content of five active components in 12 batches of samples from different different producing areas was in the range of 39.85-67.82 mg/g,and the content of swertimarin was the highest(33.87-61.44 mg/g).The PCA results showed that the variance contribution rates of the first principal component and the second principal component were 36.30%and 31.70%,respectively.The main contribution components screened were luteolin(0.6
作者
兰婷
朱贲贲
陈圆
杨鹏杰
王巍嵩
LAN Ting;ZHU Benben;CHEN Yuan;YANG Pengjie;WANG Weisong(Affiliated Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia,China 010050;Peking University Cancer Hospital Inner Mongolia Hospital·Affiliated Cancer Hospital of Inner Mongolia Medical University,Hohhot,Inner Mongolia,China 010040)
出处
《中国药业》
CAS
2023年第17期63-68,共6页
China Pharmaceuticals
基金
内蒙古自治区自然科学基金[2020MS08107]。
