HPLC-MS/MS法同时测定鸡肉中40种糖皮质激素和9种非甾体抗炎药物残留
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摘要
建立了高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定鸡胸肉中40种糖皮质激素和9种非甾体抗炎药物残留。样品用90%乙腈-水溶液(V/V,含0.1%甲酸溶液)提取,经Oasis PRIME HLB固相萃取柱净化,浓缩后采用Waters Acquity UPLC BEH C18色谱柱(2.1 mm×100 mm×1.7μm)分离,以0.1%甲酸溶液和乙腈为流动相进行梯度洗脱。采用电喷雾正离子模式,以多反应监测模式进行质谱检测。结果表明,在2～100μg/L范围内,鸡肉中49种兽药残留的线性关系良好,线性相关系数均大于0.996,回收率为60.2%～111.7%,相对标准偏差(n=6)均在1.3%～12.0%之间,检出限为0.1～0.5μg/kg,定量限为0.3～1.5μg/kg。该方法操作简单、灵敏度高、准确性好,可同时测定鸡肉中的糖皮质激素和非甾体抗炎药物残留。
A method for simultaneous determination of residues of 40 glucocorticoids and 9 non-steroidal anti-inflammatory drugs(NSAIDs) in chicken tissues by HPLC-MS/MS was developed. Glucocorticoids and NSAIDs, which were widely used to treat animal diseases and improve growth rate, were ofen fed to animals illegally. Furthermore, this misuse had caused several risks to human health as toxic effects and hypertension. In order to increase food safety, maximum residue levels(MRLs) were established for glucocorticoids and NSAIDs in animal-derived food. The establishment of a high sensitivity and accurate method for glucocorticoids and NSAIDs detection is necessary. In this study, the target compounds in chicken breast sample were extracted by 90% acetonitrile aqueous solution(V/V, containing 0.1% formic acid) and purified by the Oasis PRIME HLB, a new reversed phase SPE with only one pass-through clean-up step. Comparing with traditional solid phase extraction, the new SPE eliminates pre-equilibration and conditioning steps maked a faster workflow. In addition, it had an excellent capacity for removing matrix interferences, particularly phospholipids. The chromatographic separation was performed on a Waters Acquity UPLC BEH column(2.1 mm×100 mm×1.7 μm) and gradient eluted by 0.1% formic acid and acetonitrile and analyzed by liquid chromatography-tandem mass spectrometry in multiple reaction monitoring(MRM) mode via positive electrospray ionization.All the compounds were calibrated by the external method. To minimize the matrix effect, this experiment results were quantified by using the matrix-matched calibration curve. The results showed that the linear ranges of 49 drugs were 2-100 μg/L and the correlation coefficients were above 0.996. The recoveries were between 60.2% and 111.7% with the relative standard deviations from 1.3% to 12.0%(n=6).The limits of detection were 0.1-0.5 μg/kg, and the limits of quantification were 0.3-1.5 μg/kg, respectively. The method is easy to operate, sensitive, accurate, and is suitable for the simultaneous determination of glucocorticoids and NSAIDs residues in chicken tissues.
A method for simultaneous determination of residues of 40 glucocorticoids and 9 non-steroidal anti-inflammatory drugs(NSAIDs) in chicken tissues by HPLC-MS/MS was developed. Glucocorticoids and NSAIDs, which were widely used to treat animal diseases and improve growth rate, were ofen fed to animals illegally. Furthermore, this misuse had caused several risks to human health as toxic effects and hypertension. In order to increase food safety, maximum residue levels(MRLs) were established for glucocorticoids and NSAIDs in animal-derived food. The establishment of a high sensitivity and accurate method for glucocorticoids and NSAIDs detection is necessary. In this study, the target compounds in chicken breast sample were extracted by 90% acetonitrile aqueous solution(V/V, containing 0.1% formic acid) and purified by the Oasis PRIME HLB, a new reversed phase SPE with only one pass-through clean-up step. Comparing with traditional solid phase extraction, the new SPE eliminates pre-equilibration and conditioning steps maked a faster workflow. In addition, it had an excellent capacity for removing matrix interferences, particularly phospholipids. The chromatographic separation was performed on a Waters Acquity UPLC BEH column(2.1 mm×100 mm×1.7 μm) and gradient eluted by 0.1% formic acid and acetonitrile and analyzed by liquid chromatography-tandem mass spectrometry in multiple reaction monitoring(MRM) mode via positive electrospray ionization.All the compounds were calibrated by the external method. To minimize the matrix effect, this experiment results were quantified by using the matrix-matched calibration curve. The results showed that the linear ranges of 49 drugs were 2-100 μg/L and the correlation coefficients were above 0.996. The recoveries were between 60.2% and 111.7% with the relative standard deviations from 1.3% to 12.0%(n=6).The limits of detection were 0.1-0.5 μg/kg, and the limits of quantification were 0.3-1.5 μg/kg, respectively. The method is easy to operate, sensitive, accurate, and is suitable for the simultaneous determination of glucocorticoids and NSAIDs residues in chicken tissues.
引文
[1] WALKER D J, SPENCER K A. Glucocorticoid programming of neuroimmune function[J]. General and Comparative Endocrinology, 2018, 256: 80-88.
[2] HANSEN K E, KLEKER B, SAFDAR N, et al. A systematic review and meta-analysis of glucocorticoid-induced osteoporosis in children[J]. Seminars in Arthritis and Rheumatism, 2014, 44(1): 47-54.
[3] GROSSER T, RICCIOTTI E, FITZGERALD G A. The cardiovascular pharmacology of nonsteroidal anti-inflammatory drugs[J]. Trends in Pharmacological Sciences, 2017, 38(8): 733-748.
[4] 动物性食品中兽药最高残留限量. 中华人民共和国农业部第235号公告[EB/OL]. 2002-12-24. Http://Jiuban.moa.gov.cn/zwllm/tzgg/gg/200302/t20030226_59300.htm.
[5] 欧贝丽,赵佳丽,邹燕,等. HPLC同时测定中成药及保健品中添加的16种糖皮质激素[J]. 中国现代应用药学,2016,33(9):1 178-1 182. OU Beili, ZHAO Jiali, ZOU Yan, et al. Simultaneous determination of 16 glucocorticoids added in chinese patent medicine and health products by HPLC[J]. Chinese Journal of Modern Applied Pharmacy, 2016, 33(9): 1 178-1 182(in Chinese).
[6] RAHIM M A, IBRAHIM W A W, RAMLI Z, et al. New sol-gel hybrid material in solid phase extraction combined with liquid chromatography for the determination of non-steroidal anti-inflammatory drugs in water samples[J]. Chromatographia, 2016, 79(7/8): 421-429.
[7] 王莉,倪莹莹,王慧,等. HPLC-MS/MS法测定猪肌肉、肝脏及肾脏中双氯芬酸钠残留量[J]. 质谱学报,2015,36(3):237-242. WANG Li, NI Yingying, WANG Hui, et al. Determination of diclofenac sodium residue in swine muscle, liver or kidney by HPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2015, 36(3): 237-242(in Chinese).
[8] 龚兰,陈明,魏娴,等. 高效液相-电喷雾串联质谱法测定动物肌肉组织中的糖皮质激素[J]. 分析化学,2016,44(1):74-80. GONG Lan, CHEN Ming, WEI Xian, et al. Determination of glucocorticoids in muscle tissues by liquid chromatography-electrospray tandem mass spectrometry[J]. Chinese Journal of Analytical Chemistry, 2016, 44(1): 74-80(in Chinese).
[9] 张建丽,王占良,张亦农. LC-MS/MS 法定性检测保健品中非法添加的镇静催眠药[J]. 质谱学报,2009,30(5):275-277. ZHANG Jianli, WANG Zhanliang, ZHANG Yinong. Detection of sedative hypnotic drugs in health products by LC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2009, 30(5): 275-277(in Chinese).
[10] AMENDOLA L, GARRIBBA F, BOTRè F. Determination of endogenous and synthetic glucocorticoids in human urine by gas chromatography-mass spectrometry following microwave-assisted derivatization[J]. Analytica Chimica Acta, 2003, 489(2): 233-243.
[11] AHMAD S M, ALMEIDA C, NENG N R, et al. Bar adsorptive microextraction (BAμE) coated with mixed sorbentphases-enhanced selectivity for the determination of non-steroidalanti-inflammatory drugs in real matrices in combination with capillary electrophoresis[J]. Journal of Chromatography B, 2016, 1 008(6): 115-124.
[12] 高文惠,王姣姣,贾英民. 分子印迹固相萃取-液相色谱法测定食品中硝基呋喃类药物残留[J]. 中国食品学报,2014,14(9):183-189. GAO Wenhui, WANG Jiaojiao, JIA Yingmin. Determination of nitrofuran antibiotic residues in foods by molecularly imprinted solid phase extraction-liquid chromatography[J]. Journal of Chinese Institute of Food Science and Technology, 2014, 14(9): 183-189(in Chinese).
[13] SPIELMEYER A, AHLBORN J, HAMSCHER G. Simultaneous determination of 14 sulfonamides and tetracyclines in biogas plants by liquid-liquid-extraction and liquid chromatography tandem mass spectrometry[J]. Analytical and Bioanalytical Chemistry, 2014, 406(11): 2 513-2 514.
[14] SUN L L, MEI L Y, YANG H, et al. Development and application of immunoaffinity column for the simultaneous determination of norfloxacin, pefloxacin, lomefloxacin, and enrofloxacin in swine and chicken meat samples[J]. Food Analytical Methods, 2016, 9(2): 342-352.
[15] 王立闱,范军,吴国华,等. 在线凝胶色谱-串联三重四极杆质谱法测定大豆油中42种农药残留[J]. 质谱学报,2016,37(4):374-384. WANG Liwei, FAN Jun, WU Guohua, et al. Determination of 42 pesticide residues in soybean oil using on line GCP-GC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2016, 37(4): 374-384(in Chinese).
[2] HANSEN K E, KLEKER B, SAFDAR N, et al. A systematic review and meta-analysis of glucocorticoid-induced osteoporosis in children[J]. Seminars in Arthritis and Rheumatism, 2014, 44(1): 47-54.
[3] GROSSER T, RICCIOTTI E, FITZGERALD G A. The cardiovascular pharmacology of nonsteroidal anti-inflammatory drugs[J]. Trends in Pharmacological Sciences, 2017, 38(8): 733-748.
[4] 动物性食品中兽药最高残留限量. 中华人民共和国农业部第235号公告[EB/OL]. 2002-12-24. Http://Jiuban.moa.gov.cn/zwllm/tzgg/gg/200302/t20030226_59300.htm.
[5] 欧贝丽,赵佳丽,邹燕,等. HPLC同时测定中成药及保健品中添加的16种糖皮质激素[J]. 中国现代应用药学,2016,33(9):1 178-1 182. OU Beili, ZHAO Jiali, ZOU Yan, et al. Simultaneous determination of 16 glucocorticoids added in chinese patent medicine and health products by HPLC[J]. Chinese Journal of Modern Applied Pharmacy, 2016, 33(9): 1 178-1 182(in Chinese).
[6] RAHIM M A, IBRAHIM W A W, RAMLI Z, et al. New sol-gel hybrid material in solid phase extraction combined with liquid chromatography for the determination of non-steroidal anti-inflammatory drugs in water samples[J]. Chromatographia, 2016, 79(7/8): 421-429.
[7] 王莉,倪莹莹,王慧,等. HPLC-MS/MS法测定猪肌肉、肝脏及肾脏中双氯芬酸钠残留量[J]. 质谱学报,2015,36(3):237-242. WANG Li, NI Yingying, WANG Hui, et al. Determination of diclofenac sodium residue in swine muscle, liver or kidney by HPLC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2015, 36(3): 237-242(in Chinese).
[8] 龚兰,陈明,魏娴,等. 高效液相-电喷雾串联质谱法测定动物肌肉组织中的糖皮质激素[J]. 分析化学,2016,44(1):74-80. GONG Lan, CHEN Ming, WEI Xian, et al. Determination of glucocorticoids in muscle tissues by liquid chromatography-electrospray tandem mass spectrometry[J]. Chinese Journal of Analytical Chemistry, 2016, 44(1): 74-80(in Chinese).
[9] 张建丽,王占良,张亦农. LC-MS/MS 法定性检测保健品中非法添加的镇静催眠药[J]. 质谱学报,2009,30(5):275-277. ZHANG Jianli, WANG Zhanliang, ZHANG Yinong. Detection of sedative hypnotic drugs in health products by LC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2009, 30(5): 275-277(in Chinese).
[10] AMENDOLA L, GARRIBBA F, BOTRè F. Determination of endogenous and synthetic glucocorticoids in human urine by gas chromatography-mass spectrometry following microwave-assisted derivatization[J]. Analytica Chimica Acta, 2003, 489(2): 233-243.
[11] AHMAD S M, ALMEIDA C, NENG N R, et al. Bar adsorptive microextraction (BAμE) coated with mixed sorbentphases-enhanced selectivity for the determination of non-steroidalanti-inflammatory drugs in real matrices in combination with capillary electrophoresis[J]. Journal of Chromatography B, 2016, 1 008(6): 115-124.
[12] 高文惠,王姣姣,贾英民. 分子印迹固相萃取-液相色谱法测定食品中硝基呋喃类药物残留[J]. 中国食品学报,2014,14(9):183-189. GAO Wenhui, WANG Jiaojiao, JIA Yingmin. Determination of nitrofuran antibiotic residues in foods by molecularly imprinted solid phase extraction-liquid chromatography[J]. Journal of Chinese Institute of Food Science and Technology, 2014, 14(9): 183-189(in Chinese).
[13] SPIELMEYER A, AHLBORN J, HAMSCHER G. Simultaneous determination of 14 sulfonamides and tetracyclines in biogas plants by liquid-liquid-extraction and liquid chromatography tandem mass spectrometry[J]. Analytical and Bioanalytical Chemistry, 2014, 406(11): 2 513-2 514.
[14] SUN L L, MEI L Y, YANG H, et al. Development and application of immunoaffinity column for the simultaneous determination of norfloxacin, pefloxacin, lomefloxacin, and enrofloxacin in swine and chicken meat samples[J]. Food Analytical Methods, 2016, 9(2): 342-352.
[15] 王立闱,范军,吴国华,等. 在线凝胶色谱-串联三重四极杆质谱法测定大豆油中42种农药残留[J]. 质谱学报,2016,37(4):374-384. WANG Liwei, FAN Jun, WU Guohua, et al. Determination of 42 pesticide residues in soybean oil using on line GCP-GC-MS/MS[J]. Journal of Chinese Mass Spectrometry Society, 2016, 37(4): 374-384(in Chinese).