摘要
目的利用高效液相色谱联合质谱技术鉴定消瘀泄浊饮中的活性成分,并利用HPLC法测定毛蕊异黄酮葡萄糖苷、大黄素、芦荟大黄素、杯苋甾酮、苦杏仁苷这5种成分的含量。方法采用液质联用技术测定消瘀泄浊饮中的生物活性成分,色谱柱:Waters HSS T3(100 mm×2.1 mm,1.8μm),流动相为0.1%甲酸-水溶液→0.1%甲酸-乙腈-异丙醇,梯度洗脱;流速:0.3 mL/min;柱温:40℃。高分辨质谱:鞘气40 Arb;辅助气10 Arb;离子喷雾电压3000 V/-2800 V;温度350℃;离子传输管温度320℃;扫描模式为Full-scan MS2模式。采用Progenesis QI进行基线过滤、峰识别、积分、保留时间校正、峰对齐,对比公共数据库和实验室自建数据库后得到消瘀泄浊饮中的生物活性成分。选择Sharpsil-U C_(18)色谱柱;流动相为甲醇→超纯水;梯度洗脱;流速:1.0 mL/min;柱温:30℃;进样量:10μL;检测波长:210 nm(苦杏仁苷、杯苋甾酮),260 nm(毛蕊异黄酮葡萄糖苷、大黄素、芦荟大黄素)。结果从消瘀泄浊饮中共找到276种生物活性成分,其中负离子模式下找到119种,正离子模式下找到157种。选择毛蕊异黄酮葡萄糖苷、大黄素、芦荟大黄素、杯苋甾酮、苦杏仁苷为消瘀泄浊饮的指标成分。各成分均能有效检出,分离度良好,阴性无干扰;5种成分在测定的进样浓度范围内与峰面积线性关系良好;平均加样回收率为99.94%~101.07%;RSD为0.78%~2.27%。结论液质联用技术可以有效鉴定消瘀泄浊饮中的生物活性成分,所建立的HPLC法稳定性好,特异性强,可以为消瘀泄浊饮的质量控制和评价提供参考依据。
Objective To identify the active components in Xiaoyu Xiezhuo Yin(消瘀泄浊饮)by high performance liquid chromatography combined with mass spectrometry,and to determine the contents of calycosin glucoside,emodin,aloe emodin,cuylesterone and amygdalin by HPLC.Methods The bioactive components in Xiaoyu Xiezhuo Yin were determined by LC-MS/MS.The chromatographic column was Waters HSS T3(100 mm×2.1 mm,1.8μm),and the mobile phase was 0.1%formic acid-aqueous solution→0.1%formic acid-acetonile-isopropyl alcohol,gradient elution;Flow rate:0.3 mL/min;The column temperature was 40℃.High resolution mass spectrometry(Q Exactive,Thermo,USA);Sheath gas 40 Arb;Auxiliary gas 10 Arb;Ion spray voltage 3000 V/-2800 V;Temperature 350℃;Ion transport tube temperature 320℃;The scanning mode is the Full-scan MS2 mode.Progenesis QI was used for baseline filtering,peak identification,integration,retention time correction,and peak alignment.The bioactive components in Xiaoyu Xiezhuo Yin were obtained after comparing the public database and the self-made database.Sharpsil-U C_(18) column was selected.The mobile phase was methanol→ultrapure water.Gradient elution;Flow rate:1.0 mL/min;Column temperature:30℃;Injection volume:10μL.Detection wavelength:210 nm(amygdalin,cup amaranth),260 nm(calycosin glucoside,emodin,aloe emodin).Results A total of 276 bioactive components were found from Xiaoyu Xiezhuo Yin,of which 119 were found in negative ion mode and 157 were found in positive ion mode.Calycosin glucoside,emodin,aloe emodin,cup amaranth and amygdalin were selected as the index components of Xiaoyu Xiezhuo Yin.All components can be effectively detected with good separation and no negative interference.The five components had good linear relationship with the peak area within the measured concentration range. The average recovery rate was 99.94%-101.07%. RSD ranged from 0.78% to 2.27%. Conclusion The LC-MS/MS technology can effectively identify the bioactive components in Xiaoyu Xiezhuo Yin. The established HPLC method has g
作者
范桢亮
鲁科达
范军芬
张培培
夏虹
马红珍
FAN Zhenliang;LU Keda;FAN Junfen;ZHANG Peipei;XIA Hong;MA Hongzhen(The First Affiliated Hospital of Zhejiang Chinese Medical University,Zhejiang Provincial Hospital of Chinese Medicine,Hangzhou 310006,Zhejiang,China;The Third Affiliated Hospital of Zhejiang Chinese Medical University,Zhejiang Zhongshan Hospital,Hangzhou 310006,Zhejiang,China)
出处
《辽宁中医药大学学报》
CAS
2024年第5期32-37,共6页
Journal of Liaoning University of Traditional Chinese Medicine
基金
国家自然科学基金项目(82104759,82204999,82274432)
浙江省基础公益研究计划自然科学基金探索项目(LQ22H270002)。
关键词
消瘀泄浊饮
高效液相色谱
高分辨质谱
成分鉴定
含量测定
Xiaoyu Xiezhuo Yin(消瘀泄浊饮)
high performance liquid chromatography
high resolution mass spectrometry
component identification
determination of content
