编制说明-畜禽肉中食源性兴奋剂和孕激素的测定 液相色谱-串联质谱法

1畜禽肉中食源性兴奋剂和孕激素的测定液相色谱-串联质谱法2 4 4 4 4 52.1标准编制的必要性 52.2标准编制的预期效果 7 7 7 7 8 8 10 10 11 115.3.2混合标准储备液的配制 115.3.3外标混合标准中间液的配制 12 12 12 12 12 13 13 133 13 13 13 15 15 21 21 22 23 23 23 24 24 34 35 36 49 49 49 49 514为保障冬奥食品安全，特别是对畜禽肉供应保障PXM2020_178305_000007），52标准制定的必要性和预期效果2.1标准编制的必要性发布的《2021年禁用清单》中明确规定β2-激动剂、蛋白同化制剂、糖皮质激素类、6食源性兴奋剂等激素物质检测方法主要有色谱-质谱法、酶联免疫法、生物芯片等技术，目前色谱-质谱因为其准确度高、灵敏度高、检测通量高等优势是食源性兴奋剂等激素的主要检测手段。然而食源性兴奋剂检测依然面临较大挑战，主要有以（5）SN/T4744-2017进出口法（7）GB31658.22-2022食品7品过两种净化柱，耗时较长，需要18种内标物，检测成本较高。其他2.2标准编制的预期效果孕激素共六大类化合物为研究对象，结合QuEChERS样品前处理技术，完成了《畜同时测定45个化合物，实现了高通量和高效率，该标准方法能够为大型体育赛事运3标准制定的原则和依据在实施过程中，本项目组按照《实验室质量控制规范4主要技术内容的论据无8表1被测化合物及对应9种内标的名称、CAS号、供应来源、纯度/浓度序号目标化合物英文名称CAS供应商纯度/浓度克伦特罗21898-19-1Dr.Ehrenstorfer99.3%2.妥布特罗41570-61-0TorontoResearchChemicals97.0%班布特罗81732-46-9Dr.Ehrenstorfer98.6%4.克伦丙罗38339-11-6Dr.Ehrenstorfer98.0%喷布特罗28163-36-2WITEGALaboratorienBerlin-AdlershofGmbH99.7%克伦塞罗1435934-75-0WITEGALaboratorienBerlin-AdlershofGmbH99.8%7.马布特罗54240-36-7Dr.Ehrenstorfer90.6%溴布特罗41937-02-4WITEGALaboratorienBerlin-AdlershofGmbH99.5%9.西马特罗54239-37-1TorontoResearchChemicals98.0%氯丙那林6933-90-0Dr.Ehrenstorfer98.0%奥西那林5874-97-5Dr.Ehrenstorfer99.2%沙丁胺醇18559-94-9Dr.Ehrenstorfer99.9%119520-06-8WITEGALaboratorienBerlin-AdlershofGmbH99.6%莱克多巴胺90274-24-1StanfordAnalyticalChemicalsInc.95.8%沙美特罗89365-50-4Dr.Ehrenstorfe99.4%阿替洛尔Atenolol29122-68-7Dr.Ehrenstorfer99.0%普萘洛尔318-98-9Dr.Ehrenstorfer99.6%美托洛尔56392-17-7Dr.Ehrenstorfer98.0%98.39.40.氨苯喋啶螺内酯坎利酮睾酮诺龙去氢甲睾酮勃地酮群勃龙雄烯二酮脱氢表雄酮甲基睾酮美睾酮美雄诺龙司坦唑醇地塞米松氢化可的松波尼松龙可的松波尼松醋酸氟轻松氟氢可的松倍氯米松Triamterene396-01-0SpironolactoneCanrenone976-71-6Testosterone58-22-0Nortestosterone434-22-0Methandienone72-63-9Boldenone846-48-0Trenbolone10161-33-84-Androstene-363-5-8Prasterone53-43-0Methyltestosterone58Mesterolone1424-00-6Mestanolone521-11-9Stanozolol10418-03-8Dexamethasone50-02-2Hydrocortisone50-23-7Prednisolone50-24-8Cortisone53-06-5Prednisone53-03-267-73-2AcetonideFludrocortison127-31-1Beclomethasone4419-39-0TorontoResearchChemicalsStanfordAnalyticalChemicalsInc.TorontoResearchChemicalsDr.EhrenstorferDr.EhrenstorferStanfordAnalyticalChemicalsInc.Dr.EhrenstorferDr.EhrenstorferTorontoResearchChemicalsDr.EhrenstorferDr.EhrenstorferAChemtekInc.Dr.EhrenstorferDr.EhrenstorferDr.EhrenstorferDr.EhrenstorferDr.EhrenstorferDr.EhrenstorferDr.EhrenstorferTorontoResearchChemicalsTorontoResearchChemicalsTorontoResearchChemicals98.0%99.5%98.0%99.3%96.7%99.3%98.8%96.3%99.7%99.3%97.7%99.0%97.8%99.9%99.2%99.3%99.8%99.8%98.0%99.3%99.3%41.孕酮57-83-0Dr.Ehrenstorfer99.7%42.醋酸美伦孕酮2919-66-6Dr.Ehrenstorfer99.0%43.醋酸甲羟孕酮71-58-9Dr.Ehrenstorfer99.2%44.17α-羟孕酮604-09-1Dr.Ehrenstorfer99.2%45.甲地孕酮3562-63-8Dr.Ehrenstorfer99.4%I-1克伦特罗-D9129138-58-5StanfordAnalyticalChemicalsInc.99.7%I-2莱克多巴胺-D590274-24-1AChemtekInc.95.0%I-3沙丁胺醇-D31219798-60-3AChemtekInc.98.5%I-4去甲乌药碱-D4Methyltestosterone-2249814-83-1FirstStandard100μg/mLI-5睾酮-D377546-39-5TorontoResearchChemicals100μg/mLI-6勃地酮-D3361432-76-0AChemtekInc.99.2%I-7孕酮-D915775-74-3TorontoResearchChemicals95.0%I-8氢化可的松-D3115699-92-8AChemtekInc.10μg/mLI-9地塞米松-D5358731-91-6TorontoResearchChemicals98.0%国药集团试剂公司；N-丙基乙二胺键合固相吸附材料（PSA，40-60μm键合硅胶吸附材料（C18，40-60μm）、氨基吸附剂（NH2，40-60μm）均购于博纳艾ACQUITYUPLC超高效液相色谱仪、XevoTQ-S四级杆串联质谱仪，美国瑞诚仪器有限公司；涡旋混合器：vortex-genie2，美国s），5.3.2混合标准储备液的配制序号化合物储备液浓度(μg/mL)中间液浓度(μg/mL)组别序号化合物储备液浓度(μg/mL)中间液浓度(μg/mL)组别1克伦特罗128脱氢表雄酮22002妥布特罗129甲基睾酮22003班布特罗130美睾酮82004克伦丙罗220031美雄诺龙22005喷布特罗132司坦唑醇22006克伦塞罗133地塞米松44007马布特罗220034氢化可的松44008溴布特罗135波尼松龙44009西马特罗220036可的松4400氯丙那林137波尼松4400奥西那林138醋酸氟轻松4400沙丁胺醇139氟氢可的松4400齐帕特罗140倍氯米松4400莱克多巴胺141孕酮2200序号化合物储备液浓度(μg/mL)中间液浓度(μg/mL)组别序号化合物储备液浓度(μg/mL)中间液浓度(μg/mL)组别沙美特罗142醋酸美伦孕酮2200阿替洛尔143醋酸甲羟孕酮2200普萘洛尔14417α-羟孕酮2200美托洛尔145甲地孕酮2200氨苯喋啶2200克伦特罗-D9120螺内酯2200I-2莱克多巴胺-D5121坎利酮2200I-3沙丁胺醇-D3122睾酮2200I-4甲基睾酮-D3220023诺龙2200I-5睾酮-D3220024去氢甲睾酮2200I-6勃地酮-D3220025勃地酮2200I-7孕酮-D9220026群勃龙2200I-8氢化可的松-D3440027雄烯二酮2200I-9地塞米松-D544005.3.3外标混合标准中间液的配制称取不含被测物的阴性鸡肉样品5.0g，不加内标，按照上述方法进行前处理，表3HPLC梯度洗脱条件时间（min）A（%）B（%）流速（mL/min）09550.30.59550.3900.36.030700.38.0200.39.000.300.39550.39550.3）；）；表4目标分析物及9种内标的保留时间及主要质谱参数编号目标化合物保留时间(min)母离子(m/z)定量离子（m/z）（碰撞能）（碰撞能）碎裂电压(V)克伦特罗4.56277.4342.妥布特罗4.97228.4班布特罗368.772.2(34)294.4(20)4.克伦丙罗2.83263.326喷布特罗7.31292.5236.3(14)74.2(20)72克伦塞罗3.81319.4203.1(20)367.马布特罗5.02311.4217.2(24)237.2(14)20溴布特罗4.96367.4132.3(46)26西马特罗2.64220.4143.2(24)氯丙那林4.51214.3118.9(26)奥西那林2.20212.3125.2(20)22沙丁胺醇2.86240.4齐帕特罗2.82262.4185.2(24)202.2(18)莱克多巴胺4.21302.5107.2(26)26沙美特罗7.31416.791.2(38)25阿替洛尔2.87267.4145.2(26)74.2(22)20普萘洛尔5.94260.4美托洛尔4.75268.5氨苯喋啶4.43254.4104.2(38)20.螺内酯7.10341.5107.2(26)91.1(48)4821.坎利酮341.5107.2(28)22.睾酮7.49289.5109.2(22)4823.诺龙7.20275.5109.2(28)239.3(16)224.去氢甲睾酮7.31301.52625.勃地酮7.06287.53026.群勃龙6.98271.4199.2(22)165.3(44)6227.雄烯二酮7.19287.597.2(20)109.2(24)34编号目标化合物保留时间(min)母离子(m/z)定量离子（m/z）（碰撞能）（碰撞能）碎裂电压(V)28.脱氢表雄酮7.49289.5253.3(10)213.3(18)629.甲基睾酮7.75303.6109.2(24)3230.美睾酮8.33305.5269.4(16)95.6(32)31.美雄诺龙8.45305.6269.4(14)105.2(20)32.司坦唑醇8.33329.633.地塞米松6.78393.5373.4(8)34.氢化可的松6.42363.54435.波尼松龙6.42361.5325.4(8)2436.可的松6.19361.6163.2(24)121.2(36)37.波尼松359.52838.醋酸氟轻松6.82453.5121.2(28)433.4(8)39.氟氢可的松6.34381.5239.3(24)91.2(58)3840.倍氯米松6.81409.5373.4(8)121.2(42)641.孕酮8.40315.5109.2(22)42.醋酸美伦孕酮8.30397.2279.2(22)243.醋酸甲羟孕酮8.25387.2123.0(32)244.17α-羟孕酮7.58331.297.0(26)109.2(30)445.甲地孕酮7.87343.12I-1克伦特罗-D94.54286.4133.0(28)6I-2莱克多巴胺-D54.19307.5I-3沙丁胺醇-D32.85243.532I-4去甲乌药碱-D43.28276.5108.3（20）164.3（18）I-5睾酮-D37.49292.597.2(24)90I-6勃地酮-D37.05290.6121.2(22)I-7孕酮-D98.35324.6113.2(24)26I-8氢化可的松-D36.42366.6121.2(22)I-9地塞米松-D56.78398.6378.5(8)在最佳碎裂电压下确认目标化合物的母离子（m/z）值，然后进行二级质谱优化，通本试验被测物在正离子模式监测下具有较强的信号值大部分被测物可获得较高征离子信号重现性好，可满足监测要求，因用乙腈-水、乙腈-水（含0.1%甲酸）、甲醇-水、甲醇-水（含0.1%甲酸）、甲醇-水在ESI+模式下很容易得到准分子离子，在改善峰形之余，提高了分离效果，而甲酸本试验考察了5cm和10cm的色谱柱，发现5cm无法将分异构体美睾酮为了提高采用负离子扫描的3种化合物的检测灵敏度，试验采用单独乙腈-水体图2基质标的MRM色谱图时，随着甲醇在乙腈体系中的比例增大，其蛋白沉淀效果越来越β2-激动剂、β-阻断剂类、蛋白同化激素在酸度超过0.5%时，有基质干扰，回收率有降低的趋势；糖皮质激素、孕酮和利尿剂对提取液中乙酸的比例不敏感。当采用含0.5%乙酸的乙腈溶液作为取溶剂时，所有被测物的回收率均大于50%，确定为最优单独采用NH2时，大约有45%的化合物回收率比使用其他3种净化剂时低，且进一步采用以下4种组合方案：①PSA250mg采用组合②或③时，蛋白同化激素和糖皮质激素收率较高，但对β2-兴奋剂类和孕激综合以上多重因素，本试验最终采用组合④作为最终净化剂。其中PSA可以去除肉中的脂肪酸类和糖类干扰，C18则对脂肪和非极性化合物的吸附效果比较明显，图4采用不同净化条件时不同回收率范围的目标物个数占被测物总个数的比例图根据流动相和各目标化合物的溶解性能，考察了三种复溶溶剂：①乙腈/水（含0.1%甲酸1/9，v/v②甲醇/水（含0.1%甲酸1/9，v/v③甲醇/水（含察了目标物在基质中的信号增强或抑制的程度(signalsuppression/enhancement,增强效应，当其低于80%时显示基质抑制效应，见表5。检测发现，美托洛尔、倍方法灵敏度以检出限（LimitofDetection,LOD）和定量限（LimitofQuantitation,LOQ）来衡量。制备不同浓度的添加样品，采用本方法测定后，以定性离子对的3倍信号噪声比（S/N=3）对应的定量浓度作为检出限，10倍信号噪声），表5被测物的线性范围、相关系数（R2）、检出限（LOD）、定量限（LOQ）及对应内标化合物序号目标化合物SSE值线性范围(μg/L)线性方程相关系数（R2）LOD(μg/kg)LOQ(μg/kg)1克伦特罗克伦特罗-D997%y=1.259x+0.09820.99940.030.12妥布特罗克伦特罗-D9y=1.008x+0.01270.99920.030.13班布特罗克伦特罗-D9y=0.415x-0.05480.99950.030.14克伦丙罗克伦特罗-D90.2~20y=0.725x+0.05770.99920.070.25喷布特罗克伦特罗-D9y=4.360x+0.2590.99610.030.16克伦塞罗克伦特罗-D980%y=1.324x+0.1320.99850.030.17马布特罗克伦特罗-D90.2~20y=0.116x+0.009520.99910.070.28溴布特罗克伦特罗-D9y=0.137x-0.003380.99930.030.19西马特罗克伦特罗-D90.2~20y=1.042x+0.05990.99960.070.2氯丙那林克伦特罗-D9y=0.752x+0.02530.99970.030.1奥西那林克伦特罗-D9y=2.046x-0.07250.99930.030.1沙丁胺醇沙丁胺醇-D3y=0.432x+0.04940.99710.030.1齐帕特罗沙丁胺醇-D385%y=0.921x+0.1630.99750.030.1莱克多巴胺莱克多巴胺-D592%y=1.121x+0.3900.99890.030.1沙美特罗莱克多巴胺-D5y=0.339x-0.04200.99800.030.1阿替洛尔克伦特罗-D9y=1.789x+0.3330.99590.030.1序号目标化合物SSE值线性范围(μg/L)线性方程相关系数（R2）LOD(μg/kg)LOQ(μg/kg)普萘洛尔克伦特罗-D987%y=1.761x+0.07140.99960.03美托洛尔克伦特罗-D975%y=1.137x+0.01710.99990.03氨苯喋啶莱克多巴胺-D50.2~50y=0.998x+1.4740.99930.070.220螺内酯勃地酮-D30.2~50y=0.980x+0.1120.99920.070.221坎利酮睾酮-D399%0.2~50y=0.0116x+0.00230.99720.070.222睾酮睾酮-D398%0.2~50y=0.0428x+0.00980.99660.070.223诺龙睾酮-D30.2~50y=0.0158x+0.01050.99630.070.224去氢甲睾酮睾酮-D395%0.2~50y=0.022x+0.01110.99510.070.225勃地酮勃地酮-D385%0.2~50y=3.571x+0.9230.99710.070.226群勃龙勃地酮-D30.2~50y=2.423x+1.2300.99510.070.227雄烯二酮勃地酮-D399%0.2~50y=0.642x+0.02480.99950.070.228脱氢表雄酮勃地酮-D30.2~50y=0.135x+0.1750.99820.070.229甲基睾酮甲基睾酮-D390%0.2~50y=0.737x+0.1750.99570.070.230美睾酮甲基睾酮-D30.2~50y=0.0284+0.01520.99660.250.831美雄诺龙甲基睾酮-D395%0.2~50y=0.0274x-0.03030.99570.070.232司坦唑醇孕酮-D90.2~50y=1.858x-0.1380.99870.070.233地塞米松地塞米松-D594%0.5~200y=2.269x+0.1700.99950.160.4序号目标化合物SSE值线性范围(μg/L)线性方程相关系数（R2）LOD(μg/kg)LOQ(μg/kg)34氢化可的松氢化可的松-D399%0.5~100y=0.630x+0.0440.99910.160.435波尼松龙氢化可的松-D385%0.5~200y=0.131x+0.0.0380.99640.160.436可的松氢化可的松-D382%0.5~100y=0.894x+0.0560.99860.160.437波尼松氢化可的松-D391%0.5~100y=0.272x-0.002030.99960.160.438醋酸氟轻松氢化可的松-D380%0.5~100y=0.446x+0.05870.99340.160.439氟氢可的松氢化可的松-D381%0.5~100y=0.218x-0.006610.99990.160.440倍氯米松氢化可的松-D360%0.5~100y=0.0535x+0.01150.99920.160.441孕酮孕酮-D989%0.2~50y=0.556x+0.08870.99830.070.242醋酸美伦孕酮孕酮-D90.2~50y=0.0538x+0.00160.99980.070.243醋酸甲羟孕酮孕酮-D90.2~50y=0.0671x-0.01760.99870.070.24417α-羟孕酮睾酮-D30.2~50y=0.158x-0.05870.99400.070.245甲地孕酮甲基睾酮-D30.2~50y=0.0298x+0.00620.99760.070.2表6鸡肉加标测定结果、回收率、相对标准偏差（n=6）化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）123456克伦特罗000.080.1090.1980.430.380.450.430.440.427.3妥布特罗0.10.090.090.100.0993.37.00.40.360.440.330.420.330.410.3895.3班布特罗0.350.380.320.380.320.350.3587.77.7克伦丙罗620.380.340.380.410.380.3792.070.810.870.800.870.800.844.5喷布特罗070.180.1784.080.340.260.300.260.340.3074.7克伦塞罗0020.420.460.420.460.430.434.8马布特罗0.40.420.450.440.440.420.460.430.920.800.930.830.900.876.2溴布特罗0化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）1234560.40.420.420.430.440.390.440.424.4西马特罗60.230.1930.420.420.420.440.420.430.780.800.800.770.730.7796.04.3氯丙那林0.380.410.350.410.360.380.3895.7奥西那林0.10.080.080.080.070.060.070.0760.130.1500.2563.39.9沙丁胺醇0.10.090.090.100.090.090.100.09090.2098.000.420.410.450.390.440.425.3齐帕特罗0.10.060.070.060.060.060.060.0765.35.030.130.1366.000.290.280.300.270.300.2972.74.5莱克多巴胺000.40.410.420.420.430.420.450.423.4沙美特罗80.070.090.100.090.0950.270.320.270.310.270.300.2973.07.7阿替洛尔0.10.090.090.090.0950.150.270.260.250.260.250.270.2665.34.2化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）123456普萘洛尔000.190.300.330.270.340.290.340.3178.09.9美托洛尔0.370.380.360.380.340.350.3690.73.6氨苯喋啶90.180.1994.010.310.310.300.320.300.320.690.670.700.700.710.6885.54.8螺内酯80.200.1892.060.390.420.380.360.430.3998.030.860.880.820.830.820.843.0坎利酮80.190.200.320.370.380.320.380.380.360.700.730.740.730.700.770.7390.83.5睾酮60.130.1340.380.340.350.310.340.350.840.740.750.800.770.780.7897.54.8诺龙80.200.190.390.400.330.410.380.370.3895.000.680.710.770.710.740.7290.04.3去氢甲睾酮0.430.470.390.430.380.410.420.900.860.940.920.820.904.9勃地酮50.180.1888.7化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）1234560.40.340.340.420.340.340.320.3587.040.920.880.770.810.840.846.3群勃龙090.200.190.390.380.430.380.390.420.4099.76.00.80.870.910.900.890.820.870.883.4雄烯二酮00.40.470.420.450.390.460.340.420.80.940.930.970.920.920.900.932.6脱氢表雄酮0.390.460.460.430.460.380.450.850.850.830.860.870.85甲基睾酮50.380.380.360.350.410.410.380.750.810.880.860.740.800.816.9美睾酮0.80.520.560.510.480.470.580.5265.08.0274.14.842.552.702.622.672.742.622.6566.32.5美雄诺龙0.40.440.360.420.460.470.380.420.80.810.720.730.700.650.690.7289.57.4司坦唑醇80.210.1994.00.40.340.410.360.420.380.350.380.870.730.850.860.770.860.827.1地塞米松0.40.570.460.410.460.460.400.460.80.850.770.730.820.810.830.805.5化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）12345693.88.2氢化可的松0.40.320.300.340.290.320.340.320.650.730.740.780.800.800.7593.77.884.53.6波尼松龙80.330.290.260.340.2980.610.620.580.580.670.6176.05.98.8可的松0.40.410.440.430.430.440.440.490.890.620.670.740.620.6986.092.26.8波尼松0.40.340.310.380.380.360.390.360.530.660.600.630.570.560.5973.893.14.3醋酸氟轻松0.40.320.380.440.390.400.380.3910.710.610.670.630.630.6480.56.385.74.9氟氢可的松0.40.400.390.440.410.410.420.450.760.720.780.780.660.7390.78.093.35.6倍氯米松0.40.340.330.360.390.380.320.3588.00.80.840.790.810.720.730.760.7796.891.8孕酮60.160.1892.00.40.340.330.300.320.330.350.3382.06.00.80.710.710.770.700.620.660.7087.27.0化合物加标水平(μg/kg)重复数平均值(μg/kg)回收率（%）RSD（%）123456醋酸美伦孕酮00.470.400.410.420.400.500.400.760.770.820.930.810.838.4醋酸甲羟孕酮30.270.300.260.290.400.390.3310.830.930.940.870.930.904.717α-羟孕酮0.440.470.470.470.380.430.470.700.820.880.740.780.808.9甲地孕酮0.450.470.400.420.380.380.440.680.730.700.790.770.7594.07.9按照本方法测定了9个肉样品，并采用国家标准方法《GB/T用国标方法验证了20个与本研究一致的化合物：群勃龙、勃地酮、诺龙、雄烯表7肉类样品采用本方法检测结果及国标法验证结果样品名称本方法检测结果(μg/kg）国标方法检测结果(μg/kg）氢化可的松可的松雄烯二酮氢化可的松可的松雄烯二酮前尖（猪肉）4.90-5.21-瘦肉（猪肉）7.74-8.07-后尖（猪肉）5.600.96-6.71-鸡胸------鸡翅中------鸭肉------牛腿肉5.960.42-6.320.45-牛腱子3.800.263.44-牛腩5.762.36-5.74-定量限(μg/kg）注：表中“-”代表低于定量限。标方法过程繁琐，消耗试剂耗材多，方法测定时间长，表8两种方法差值的统计学分析样品名称d值（d=C1-C2）氢化可的松可的松猪肉1-0.310猪肉2-2.00-0.33猪肉3-0.42牛肉1-0.36-0.03牛肉20.360.17牛肉30.020.37-0.57-0.04差值标准差（Sdi）0.860.30差值平均标准差（S=Sdi/“）0.350.33采用本方法检测了12个牛肉样品，结果检出4个化合物，结果见表9。表9牛肉样品的检测结果样品名称检测结果(μg/kg)氢化可的松可的松雄烯二酮睾酮牛腿肉1号牛2.0-0.8牛腿肉2号牛22.6--牛腿肉3号牛22.22.0--牛腿肉4号牛-牛腿肉5号牛-牛腿肉6号牛3.3---牛腿肉7号牛3.3---牛腿肉8号牛5.5---牛腿肉9号牛7.6---牛腿肉10号牛7.6--霖肉（牛）0.6牛前部肉0.62家参与验证的机构分别为北京市疾病预防控制中心（A）和北京市延庆区疾病预防控制中心（B）。验证结果如下。）；表10实验室A的验证结果化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值1.克伦特罗0.10.080.100.080.090.080.090.0960.1888.00.40.400.440.420.530.420.530.462.妥布特罗0070.420.390.510.380.500.433.班布特罗0.40.420.410.350.490.350.470.424.克伦丙罗0.430.380.420.370.420.340.398.660.740.740.890.720.900.7998.85.喷布特罗090.340.260.360.270.380.3279.76.克伦塞罗00.420.380.400.440.400.450.416.07.马布特罗20.400.400.340.370.400.397.660.880.830.920.820.940.885.78.溴布特罗60.090.090.060.090.0879.883.3化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值0.40.420.440.400.460.350.420.429.29.西马特罗30.380.410.420.430.410.410.80.690.780.810.820.820.840.796.899.010.氯丙那林0.40.380.410.340.460.350.450.4011.奥西那林0.10.080.070.060.060.090.060.0772.066.040.220.270.220.290.2562.012.沙丁胺醇0.10.0005.396.0096.500.420.390.510.390.500.4413.齐帕特罗0.10.070.080.060.060.070.070.078.530.139.868.00.40.290.300.270.340.270.340.309.875.014.莱克多巴胺0000.440.420.480.430.480.447.215.沙美特罗0574.00.40.260.320.280.380.260.400.3279.316.阿替洛尔90.080.100.090.0976.00.40.270.280.250.370.250.350.2973.7化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值17.普萘洛尔00.100.006.264.00.40.280.340.290.400.300.430.3484.718.美托洛尔00.40.380.410.370.460.370.470.4119.氨苯喋啶80.170.1680.070.250.280.260.290.270.2700.670.640.820.660.740.6986.020.螺内酯00.180.1892.040.400.340.350.370.380.376.350.740.730.780.800.840.775.396.721.坎利酮80.180.1888.060.340.330.340.330.370.344.986.00.80.680.600.620.710.700.660.666.783.022.睾酮20.420.410.420.450.410.400.80.690.740.710.700.740.740.723.389.823.诺龙30.320.340.360.320.360.345.580.580.550.670.590.660.618.375.724.去氢甲睾酮10.190.270.450.420.370.480.420.430.860.790.850.740.820.7897.825.勃地酮50.167.778.70.2化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值0.40.410.350.380.360.430.340.3895.00.80.780.800.740.900.780.900.828.426.群勃龙40.380.410.370.420.340.379.270.760.660.800.690.860.759.794.327.雄烯二酮90.130.390.270.340.320.380.330.3460.650.660.860.770.820.7593.828.脱氢表雄酮60.150.190.440.420.340.360.460.410.400.80.740.860.720.830.780.860.807.899.729.甲基睾酮60.340.330.320.290.330.300.325.710.790.860.920.690.900.8330.美睾酮0.80.840.900.880.740.790.890.847.30.822.332.292.182.0943.662.793.293.133.079.531.美雄诺龙20.220.200.290.260.240.269.365.00.80.700.620.700.820.660.780.7189.032.司坦唑醇0.40.420.410.430.380.450.420.460.890.830.740.830.780.826.433.地塞米松40.260.250.290.280.270.780.820.820.540.800.790.7695.0化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值7.234.氢化可的松0.40.270.260.260.270.250.350.280.650.490.500.500.360.500.5062.585.835.波尼松龙0.40.340.300.290.340.340.370.330.850.820.860.840.780.860.833.96.791.736.可的松0.40.450.420.370.340.500.390.440.900.900.710.740.930.822.837.波尼松0.40.370.340.290.300.420.350.350.620.810.860.720.710.870.7695.56.938.醋酸氟轻松0.40.290.340.290.250.260.290.290.610.560.560.540.560.620.575.271.881.139.氟氢可的松0.40.310.280.260.250.310.250.280.770.880.660.670.690.870.7694.791.940.倍氯米松0.40.300.330.330.350.380.370.348.70.670.660.620.650.620.630.643.580.396.741.孕酮80.430.420.390.410.380.4040.780.660.830.690.780.758.593.3化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值42.醋酸美伦孕酮0.40.440.420.370.450.400.380.418.00.80.580.620.630.660.700.690.657.280.843.醋酸甲羟孕酮10.20.460.440.470.380.430.380.440.910.870.860.950.900.913.844.17α-羟孕酮60.180.190.340.410.370.300.440.360.3730.760.580.740.790.770.7188.845.甲地孕酮60.420.330.320.420.390.3970.740.730.760.930.940.83表11实验室B的验证结果化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值1.克伦特罗00.420.430.480.440.470.450.455.02.妥布特罗0093.240.380.350.440.400.420.399.697.03.班布特罗070.360.340.440.380.460.3998.04.克伦丙罗0.40.380.320.420.370.420.380.389.995.00.80.800.790.770.870.860.800.814.95.喷布特罗0.10.080.090.100.080.100.080.0988.060.1380.340.280.380.320.380.3382.76.克伦塞罗0.10.080.100.080.090.1020.360.420.410.460.380.418.77.马布特罗10.180.190.380.320.370.360.420.460.3896.00.80.800.900.740.900.870.940.868.58.溴布特罗00.070.090.070.0990.70.1化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值094.00.40.380.420.340.450.420.460.419.西马特罗0.40.390.430.470.440.450.400.437.00.80.710.710.720.820.830.760.767.395.010.氯丙那林0.40.380.390.370.460.420.450.418.711.奥西那林0880.270.230.290.260.300.278.368.012.沙丁胺醇0.10.000.100.090.094.480.196.400.440.410.470.420.480.447.813.齐帕特罗0.10.070.060.080.060.070.060.079.435.664.00.40.290.310.280.380.290.380.3280.314.莱克多巴胺00.450.440.420.510.430.500.467.915.沙美特罗30.139.470.00.40.330.340.300.450.290.430.3689.316.阿替洛尔70.070.080.090.080.0881.30.1化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值50.157.070.00.40.220.280.310.2869.017.普萘洛尔0.10.00030.1330.330.300.440.280.400.3586.718.美托洛尔0.10.060.380.380.430.390.450.408.699.719.氨苯喋啶30.1568.040.240.290.260.270.260.267.265.00.80.620.640.620.690.620.680.645.080.520.螺内酯00.480.350.340.340.340.330.3691.00.80.660.780.780.700.780.700.736.891.521.坎利酮10.320.320.290.360.300.328.079.00.80.660.830.770.740.670.660.729.690.522.睾酮0.300.300.350.380.340.390.3470.740.660.730.690.720.704.487.723.诺龙59.60.270.300.350.320.270.280.3040.780.860.720.800.780.786.597.524.去氢甲睾酮化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值0.40.470.390.440.440.400.500.400.750.890.860.740.850.838.525.勃地酮88.040.350.420.410.400.380.4030.860.870.900.870.800.864.026.群勃龙80.180.160.370.340.370.400.420.350.378.010.900.760.910.750.8527.雄烯二酮0.440.460.460.470.450.420.490.940.940.940.890.940.922.828.脱氢表雄酮00.320.300.270.420.390.350.3410.650.580.670.660.750.659.381.729.甲基睾酮30.330.330.320.320.320.300.323.740.780.740.860.790.830.796.098.730.美睾酮0.80.550.570.530.560.510.510.544.667.30.828.088.743.083.193.653.533.083.277.681.931.美雄诺龙0.20.270.20.40.350.350.370.420.340.370.30.630.660.590.710.700.630.656.881.732.司坦唑醇30.260.210.2化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值0.40.340.390.380.380.360.420.387.395.00.80.790.740.680.750.700.720.735.491.533.地塞米松90.280.260.300.220.270.630.660.510.620.620.620.618.576.57.784.134.氢化可的松20.340.350.310.410.330.660.620.550.720.540.540.6175.74.735.波尼松龙0.40.450.350.420.340.350.390.3896.070.720.700.660.680.730.692.736.可的松0.40.420.440.420.470.370.460.40.580.800.730.890.790.860.7796.74.937.波尼松0.40.380.360.320.330.390.440.370.660.820.780.780.710.850.7695.54.595.838.醋酸氟轻松0.40.260.260.340.290.370.340.30.490.500.520.560.580.660.5569.27.539.氟氢可的松0.40.460.380.450.460.360.420.480.610.620.640.620.580.614.375.87.340.倍氯米松0.40.260.360.260.279.466.30.4化合物加标水平(μg/kg)测定结果(μg/kg)RSD（%）回收率（%）定量限(μg/kg)123456平均值0.80.540.580.500.510.570.530.545.867.33.269.841.孕酮10.20.360.340.300.300.260.320.3178.00.80.820.870.740.890.730.880.828.942.醋酸美伦孕酮0.450.340.410.310.360.460.3997.00.80.620.680.620.700.710.630.666.482.743.醋酸甲羟孕酮00.150.1890.040.350.420.390.400.330.379.980.900.940.930.900.860.903.344.17α-羟孕酮60.170.1574.090.360.390.340.320.400.3520.910.820.900.840.890.874.745.甲地孕酮00.260.360.420.330.400.3620.790.730.830.810.860.797.098.76采用国际标准的程度与水平的简要说明无7标准涉及的专利及相关知识产权说明本标准涉及一个发明专利，该专利已通过初审，专8重大意见分歧的处理经过和依据无9与现行法律法规和强制性标准的关系《食品安全国家标准食品中兽药最大残留限量》（GB），表12《体育局反兴奋剂字[2021]584食源性兴奋剂防控工作指南清单》中附录“建议检测的各类药物名单”及本方法的检出限药物名控制限量参考检验方法*参考检出限**(μg/kg）本方法检出限(μg/kg)1.克仑特罗不得检出SN/T1924-2011《进出口动物源食品中克仑特罗、莱克多巴胺、沙丁胺醇和特布他林残留量的测定液相色谱-质谱/质谱法》0.050.032.沙丁胺醇不得检出物源性食品中β-受体激动剂残留检测液相色谱-串联质谱法》0.250.033.莱克多巴胺不得检出0.250.034.妥布特罗不得检出0.250.035.喷布特罗不得检出0.250.036.西马特罗不得检出0.250.077.沙美特罗不得检出0.50.038.克仑丙罗不得检出0.20.079.普萘洛尔不得检出SN/T2624-2010《动物源性食品中多种碱性药物残留量的检测方法液相色谱-质谱/质谱法》0.50.0310.阿替洛尔不得检出0.0311.美托洛尔不得检出0.50.03不得检出农业部1031号公告-1-2008《动物源性食品中11种激素残留检测液相色谱-串联质谱法》0.10.0713.司坦唑醇不得检出0.10.07不得检出0.10.07不得检出0.50.0716.勃地酮不得检出0.40.0717.群勃龙不得检出0.50.07≤2μg/kg0.0719.脱氢表雄酮≤2μg/kgGB/T21981-2008《动物源食品中激素多残留检测方法液相色谱-质谱/质谱法》2.00.220.齐帕特罗不得检出SN/T2624-2010《动物源性食品中多种碱性药物残留量的检测方法液相色谱-质谱/质谱法》0.50.0321.泼尼松不得检出农业部1031号公告-2-2008《动物源性食品中糖皮质激素类药物多残留检测液相色谱-串联质谱法；GB/T21981-2008《动物源食品中激素多残留检测方法液相色谱-质谱/质谱法》0.50.222.泼尼松龙不得检出0.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