Q-Orbitrap高分辨质谱快速筛查及定量分析改善睡眠类保健品中非法添加的18个药物成分
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摘要
目的:建立高效液相色谱-四极杆/静电场轨道阱质谱法(UPLC-Q-Orbitrap)快速筛查及定量分析改善睡眠类保健品中常见非法添加的18个药物成分。方法:保健品样品以甲醇为提取溶剂进行超声提取,离心取上清后,采用Waters HSS T3色谱柱(100 mm×2.1 mm,1.8μm)进行分离,乙腈(含0.1%甲酸)和水(含2 mmol·L~(-1)乙酸铵,0.2%甲酸)为流动相,梯度洗脱(0～1 min,35%B;1～7 min,35%B→85%B;7～7.5 min,85%B→35%B;7.5～10 min,35%B);流速为0.3 mL·min~(-1);柱温:35℃。质谱采用正离子和负离子切换扫描,全扫描/数据依赖的二级扫描(Full MS/dd-MS2)模式,在1个分析周期(10 min)内完成对样品中分析物的分离和一级、二级质谱扫描。通过比较样品与对照品的色谱保留时间、一级质谱和二级质谱,确定样品中是否掺杂了化学药物,并对阳性样品进行定量分析。结果:建立的UPLC-Q-Orbitrap方法简便快捷,定性能力强,非常适合高通量非法添加筛查。18个化学药物在0.15～300 ng·mL~(-1)范围内具有良好的线性(线性相关系数均大于0.993),各类药物的检测下限为0.05～20 ng·mL~(-1),回收率为72.2%～118.7%,RSD为2.4%～9.2%。结论:该方法操作简单,分析时间短,灵敏度高,准确可靠,为打击日益猖獗的非法添加行为提供了新的手段。
Objective:To establish a Q-Orbitrap high resolution mass spectrometry method for the screening,confirmation and quantitation of 18 kinds of illegally added drugs in improving sleep health foods.Methods:The health food samples were extracted using methanol by sonication.After centrifugation,the supernatants were separated on a Waters HSS T3 column(100 mm×2.1 mm,1.8 μm).Acetonitrile containing0.1% formic acid(B)and water containing 2 mmol·L~(-1) ammonium acetate combined with 0.2% formic acid(A)were used as mobile phase by gradient elution(0~(-1) min,35%B;1-7 min,35%B → 85%B;7-7.5 min,85%B → 35%B;7.5~(-1)0 min,35%B)at a flow rate of 0.3 mL·min~(-1),and the column temperature was 35 ℃.The analytes were detected by both positive and negative ion ESI in Full MS/dd-MS2 mode.The accurate mass and MS2 fragment ions were simultaneously attained within an analysis cycle(10 min),while the potential positive samples were quantified.The chromatographic retention time,precursor ions and product ions were compared with the standard,Samples that illegally adulterated were identified.Results:The established Q-Orbitrap method possessed powerful qualitative ability and high sensitivity.The results indicated that the linearity of all the 18 kinds of drugs ranged from 0.15 ng·mL~(-1) to 300 ng·mL~(-1) with the correlation coefficients of greater than 0.993.The limits of detection were confirmed by spiked samples,and were between 0.05 and 20 ng·mL~(-1) for the 18 drugs.The recoveries were in the range of 72.2% to 118.7%,with the relative standard deviations in the range of 2.4% to 9.2%.Conclusion:This new Q-Orbitrap method was rapid,accurate and high selective,which provides a novel powerful tool for the combating with rampant illegal adulteration behaviors.
Objective:To establish a Q-Orbitrap high resolution mass spectrometry method for the screening,confirmation and quantitation of 18 kinds of illegally added drugs in improving sleep health foods.Methods:The health food samples were extracted using methanol by sonication.After centrifugation,the supernatants were separated on a Waters HSS T3 column(100 mm×2.1 mm,1.8 μm).Acetonitrile containing0.1% formic acid(B)and water containing 2 mmol·L~(-1) ammonium acetate combined with 0.2% formic acid(A)were used as mobile phase by gradient elution(0~(-1) min,35%B;1-7 min,35%B → 85%B;7-7.5 min,85%B → 35%B;7.5~(-1)0 min,35%B)at a flow rate of 0.3 mL·min~(-1),and the column temperature was 35 ℃.The analytes were detected by both positive and negative ion ESI in Full MS/dd-MS2 mode.The accurate mass and MS2 fragment ions were simultaneously attained within an analysis cycle(10 min),while the potential positive samples were quantified.The chromatographic retention time,precursor ions and product ions were compared with the standard,Samples that illegally adulterated were identified.Results:The established Q-Orbitrap method possessed powerful qualitative ability and high sensitivity.The results indicated that the linearity of all the 18 kinds of drugs ranged from 0.15 ng·mL~(-1) to 300 ng·mL~(-1) with the correlation coefficients of greater than 0.993.The limits of detection were confirmed by spiked samples,and were between 0.05 and 20 ng·mL~(-1) for the 18 drugs.The recoveries were in the range of 72.2% to 118.7%,with the relative standard deviations in the range of 2.4% to 9.2%.Conclusion:This new Q-Orbitrap method was rapid,accurate and high selective,which provides a novel powerful tool for the combating with rampant illegal adulteration behaviors.
引文
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[5]黎银波,孟庆玉,钟庆元,等.UPLC-MS/MS法快速检测减肥类保健食品中7种违法添加的药物[J].中南药学,2012,10(10):752LI YB,MENG QY,ZHONG QY,et al.Rapid identification of 7drugs illegally added into weight reducing health foods by UPLC-MS/MS[J].Cent South Pharm,2012,10(10):752
[6]屠婕红,盛静,丁宝月,等.UPLC-MS/MS法测定中成药及保健品中添加的16种降糖类化学药物[J].药物分析杂志,2014,34(10):1823TU JH,SHENG J,DING BY,et al.Determination of 16 antidiabetic chemicals added to traditional Chinese medicines and health care products by UPLC-MS/MS[J].Chin J Pharm Anal,2014,34(10):1823
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[8]巩丽萍,田晨颖,郭常川,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱快速筛查及定量分析中成药或保健品中非法添加的24种消炎止痛类化学成分[J].中国药学杂志,2015,50(24):2154GONG LP,TIAN CY,GUO CC,et al.Rapid screening and quantitative detection of 24 illegally added anti-inflammatory painkillers in traditional Chinese medicines and health foods by ultra performance liquid chromatography-quadrupole/electrostaticfield orbitrap high resolution mass spectrometry[J].Chin Pharm J,2015,50(24):2154
[9]SHI F,GUO C,GONG L,et al.Application of a high resolution benchtop quadrupole-Orbitrap mass spectrometry for the rapid screening,confirmation and quantification of illegal adulterated phosphodiesterase-5 inhibitors in herbal medicines and dietary supplements[J].J Chromatogr A,2014,1344(6):91
[10]董喆,李梦怡,吴迪,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法快速筛查及定量分析减肥类保健品中20种非法添加药物[J].食品安全质量检测学报,2017,8(10):4031DONG Z,LI MY,WU D,et al.Rapid screening and quantitative detection of 20 illegally added drugs in diet health foods by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry[J].J Food Safe Qual,2017,8(10):4031
[11]国家食品药品监督管理局药品检验补充方法和检验项目批准件(批准件编号:2009024).安神类中成药[Z].2009-12-04The Approved Compensation Process and Inspecting Item of China Food and Drug Administration(Serial number:2009024).Sedative Chinese Patent Medicines[Z].2009-12-04
[12]国家食品药品监督管理局药品检验补充方法和检验项目批准件(批准件编号:2012004).安神类中成药和保健品[Z].2012-08-02The Approved Compensation Process and Inspecting Item of China Food and Drug Administration(Serial number 2012004).Sedative Chinese Patent Medicines and Health Food[Z].2012-08-02
[2]施亚琴,姚静,张启明,等.薄层色谱法快速筛查降血糖中成药及保健食品中添加磺酰脲类化学降糖药[J].药物分析杂志,2007,27(1):36SHI YQ,YAO J,ZHANG QM,et al.TLC method for screening sulphonylureas in antidiabetics Chinese traditional patent medicines and health supplementary foods[J].Chin J Pharm Anal,2007,27(1):36
[3]王豆,杨欣,程怡凡,等.高效液相色谱法同时鉴定保健胶囊食品中非法添加的9种降糖药成分[J].药物分析杂志,2013,33(8):1377WANG D,YANG X,CHENG YF,et al.Identification of nine components of antidiabetic medicine to health food capsules by HPLC[J].Chin J Pharm Anal,2013,33(8):1377
[4]王贞媛,李丹,文红梅,等.LC-MS/MS法快速筛选保健食品中非法添加的18个止咳平喘化学成分[J].中华中医药学刊,2012,30(1):172WANG ZY,LI D,WEN HM,et al.Screening for 18 mixed illegally cough-relieving chemical composition in health food by LC-MS/MS[J].Chin Arch Tradit Chin Med,2012,30(1):172
[5]黎银波,孟庆玉,钟庆元,等.UPLC-MS/MS法快速检测减肥类保健食品中7种违法添加的药物[J].中南药学,2012,10(10):752LI YB,MENG QY,ZHONG QY,et al.Rapid identification of 7drugs illegally added into weight reducing health foods by UPLC-MS/MS[J].Cent South Pharm,2012,10(10):752
[6]屠婕红,盛静,丁宝月,等.UPLC-MS/MS法测定中成药及保健品中添加的16种降糖类化学药物[J].药物分析杂志,2014,34(10):1823TU JH,SHENG J,DING BY,et al.Determination of 16 antidiabetic chemicals added to traditional Chinese medicines and health care products by UPLC-MS/MS[J].Chin J Pharm Anal,2014,34(10):1823
[7]ZUBAREV RA,MAKAROV A.Orbitrap mass spectrometry[J].Anal Chem,2013,85(11):5288
[8]巩丽萍,田晨颖,郭常川,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱快速筛查及定量分析中成药或保健品中非法添加的24种消炎止痛类化学成分[J].中国药学杂志,2015,50(24):2154GONG LP,TIAN CY,GUO CC,et al.Rapid screening and quantitative detection of 24 illegally added anti-inflammatory painkillers in traditional Chinese medicines and health foods by ultra performance liquid chromatography-quadrupole/electrostaticfield orbitrap high resolution mass spectrometry[J].Chin Pharm J,2015,50(24):2154
[9]SHI F,GUO C,GONG L,et al.Application of a high resolution benchtop quadrupole-Orbitrap mass spectrometry for the rapid screening,confirmation and quantification of illegal adulterated phosphodiesterase-5 inhibitors in herbal medicines and dietary supplements[J].J Chromatogr A,2014,1344(6):91
[10]董喆,李梦怡,吴迪,等.超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法快速筛查及定量分析减肥类保健品中20种非法添加药物[J].食品安全质量检测学报,2017,8(10):4031DONG Z,LI MY,WU D,et al.Rapid screening and quantitative detection of 20 illegally added drugs in diet health foods by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry[J].J Food Safe Qual,2017,8(10):4031
[11]国家食品药品监督管理局药品检验补充方法和检验项目批准件(批准件编号:2009024).安神类中成药[Z].2009-12-04The Approved Compensation Process and Inspecting Item of China Food and Drug Administration(Serial number:2009024).Sedative Chinese Patent Medicines[Z].2009-12-04
[12]国家食品药品监督管理局药品检验补充方法和检验项目批准件(批准件编号:2012004).安神类中成药和保健品[Z].2012-08-02The Approved Compensation Process and Inspecting Item of China Food and Drug Administration(Serial number 2012004).Sedative Chinese Patent Medicines and Health Food[Z].2012-08-02