TCIQA-养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定 高效液相色谱-串联质谱法编制说明

P/CIQA-104-2022PAGE5中国出入境检验检疫协会团体标准《养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定高效液相色谱-串联质谱法》编制说明（征求意见稿）标准起草组二〇二四年二月《养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定高效液相色谱-串联质谱法》编制说明标准起草的基本情况（一）任务来源2022年9月15日,根据中国出入境检验检疫协会综合质量服务标准化技术委员（CIQA/TC12）立项审查结果，由中国检验认证集团广西有限公司牵头，自然资源部第四海洋研究所、北海海关综合技术服务中心参与《养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定高效液相色谱-串联质谱法》的项目制定工作，项目编号为P/CIQA-104-2022。为保证标准制订的科学性、适用性，中国检验认证集团广西有限公司和自然资源部第四海洋研究所、北海海关综合技术服务中心成立标准起草工作组，制定工作方案、明确职责分工、时间进度和有关工作要求。（二）主要起草单位、起草人及分工主要起草单位：中国检验认证集团广西有限公司、自然资源部第四海洋研究所、北海海关综合技术服务中心。主要起草人：梁光纤、张丽、梁任佳、刘冬灵、黄芯语、金其扬、陆媛苑、樊春燕、刘丹（三）简要起草过程2022年9月，由中国检验认证集团广西有限公司牵头，自然资源部第四海洋研究所、北海海关综合技术服务中心参与《养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定高效液相色谱-串联质谱法》的项目制定工作。2022年10月，中国检验认证集团广西有限公司和自然资源部第四海洋研究所、北海海关综合技术服务中心成立标准起草工作组，开始制定实验方案，经过前期大量文献调查和在实验室研究工作的基础上探索研究了检测方法条件。2022年11月-2023年9月，开始开展仪器条件研究、前处理步骤、线性范围、方法回收率、准确度、精密度等一系列技术内容的确定研究。2023年10月形成标准检测方法草案。2023年11月-12月，在完成实验室内方法学验证的基础上，组织了自然资源部第四海洋研究所、北海海关综合技术服务中心、青岛海关技术中心对本标准进行了实验室间验证，验证结果表明方法的各项技术指标满足养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定的要求。2024年1月-2月针对各家分析实际操作时标准文本中存在的问题及建议，对标准进行了修改，得到标准起草小组的一致认可，形成该标准征求意见稿和标准编写报告。三、标准的重要内容本文件对标准方法的原理、检测项目、分析步骤、线性范围、试剂和材料、仪器分析条件的优化和选择、方法检出限及定量限、回收率、正确度和精密度等重要内容进行了规定和说明。原理水样中的硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物经亲水亲脂平衡型固相萃取柱直接富集净化后，用高效液相色谱-串联质谱法测定，三苯甲烷类采用内标法定量，其他药物采用外标法定量。检测项目本标准起草小组根据常见的监测检测兽药类别，参考GB31658.17-2021《食品安全国家标准动物性食品中四环素类、磺胺类和喹诺酮类药物残留量的测定液相色谱－串联质谱法》、GB31660.1-2019《食品安全国家标准水产品中大环内酯类药物残留量的测定液相色谱-串联质谱法》等相关兽药残留检测食品安全国家标准，对硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物等7类兽药的检测项目进行了预先设定，结合仪器检测条件参数研究中对混合标准物质进行测定，重点对流动相、色谱柱、洗脱梯度进行了选择优化，确认最终的目标兽药检测项目为硝基呋喃类(呋喃唑酮、呋喃它酮)、氯霉素类（氯霉素、氟苯尼考）、三苯甲烷类（孔雀石绿、隐性孔雀石绿）、喹诺酮类（环丙沙星、二氟沙星、依诺沙星、恩诺沙星、氟甲喹、洛美沙星、诺氟沙星、氧氟沙星、培氟沙星、沙拉沙星、噁喹酸）、磺胺类（磺胺苯酰、乙酰磺胺、磺胺氯哒嗪、磺胺嘧啶、磺胺甲噁唑、磺胺间二甲氧嘧啶、磺胺邻二甲氧嘧啶、磺胺甲嘧啶、磺胺二甲嘧啶、磺胺甲噻二唑、磺胺苯吡唑、磺胺吡啶、磺胺噻唑、磺胺二甲异噁唑）、四环素类（四环素、金霉素、多西环素）和大环内酯类（红霉素、克拉霉素、阿奇霉素、吉他霉素、交沙霉素、替米考星、泰乐菌素）。试剂纯度和级别的选择除非另有规定，本方法所使用试剂均为优级纯，水为GB/T6682规定的一级水。7类兽药的养殖用水前处理和仪器检测条件的优化4.1、前处理条件本标准起草小组对根据既有参考文献及相关兽药检测的食品安全国家标准的研究，前处理关键在于样品的浓缩富集，本标准采用加标回收的方式直接比较了HBL、C18、中性氧化铝柱等3种固相萃取小柱，以及甲醇、1%0.1%甲酸乙睛、0.1%甲酸酸乙睛+甲醇等3种洗脱液的洗脱能力。结果表明，三种的洗脱能力顺序为：0.1%甲酸乙睛+甲醇＞0.1%甲酸乙睛＞甲醇；采用C18固相萃取小柱，7类兽残的回收率基本＜20%；采用中性氧化铝柱，仅有孔雀石绿可富集洗脱;采用HLB作固相萃取小柱时，7类兽药基本能有效的富集洗脱，且大部分物质满足回收率在60%-120%。故采用HBL固相萃取小柱作进行浓缩富集，并以0.1%甲酸乙睛+甲醇经行洗脱。并最终建立了以下分析步骤：水样经0.45μm滤膜过滤后，移取50mL水样，加入100μL混合内标工作溶液(5.4.5)，超声2min，吸取全部液体过固相萃取柱（预先用6mL甲醇，6mL水活化），待试样完全流出后用6mL0.1%甲酸水淋洗，弃去水淋液，抽干，再依次用6mL甲醇、6mL0.1%

甲酸乙睛洗脱，收集洗脱液于15mL

玻璃试管中，45℃水浴氮气吹干，再用1.00mL0.1%甲酸水+乙睛(95+5)溶液溶解残渣，经0.22µm滤膜过滤，待上机测定。4.2、测定条件的优化和选择液相色谱串联质谱分析条件，是养殖用水中多种类型兽药实现统一分析检测的关键，综合考虑响应值、峰形、稳定性、准确度、仪器分析时间等因素，排除在同时测定过程中被干扰影响的物质，在对测定条件反复试验与优化的基础之上，本标准对养殖用水中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物残留量的测定条件详细了规定，确立了可以进行统一分析检测的检测项目，给出了高效液相色谱-串联质谱的参考条件。4.2.1、液相色谱工作条件本标准参考既往研究选用安捷伦的EclipseXDB-C18、EclipsePlusC18（100mm×2.1mm，1.8μm）作为色谱柱，选用0.1%甲酸水溶液与乙腈作为流动相，并采用梯度洗脱的方法，可以实现7类目标兽药物质的有效分离，并将仪器分析检测时间控制在15分钟，保留时间详见表3。建立的液相色谱工作条件如下：色谱柱：C18(100mm×2.1mm，1.8μm）；柱温：35℃；进样量：10μL；流速：0.5mL/min；流动相：A为0.1%甲酸水溶液，B为乙腈，梯度洗脱条件见表1。流动相洗脱条件时间min0.1%甲酸水溶液%乙腈%09820.598228020470307595959511010013.5010015982169824.2.2、质谱工作条件本标准为实现7类兽药物质的统一分析，采用正离子、负离子同时扫描的方式，质谱分析条件主要为定性离子对、定量离子对、碰撞能量等分析参数的优化与建立。根据本实验室的仪器特性，利用仪器自带的优化软件，建立了7类兽药目标物质的质谱分析参数。建立的质谱工作条件如下：离子源：电喷雾离子源；扫描方式：正离子、负离子扫描；检测方式：多反应检测；气体温度：300℃；雾化器压力：45psi；干燥气流量：氮气11L/min；鞘流气温度：350°C；鞘流气流量：氮气11L/min；毛细管电压：3500V（正离子）；4000V（负离子）；定性离子对、定量离子对、碰撞能量见表2。定性离子对、定量离子对、碰撞能量、扫描方式药物名称母离子m/z子离子m/z碰撞能量eV扫描方式呋喃它酮325.1281.216+325.1252.2*20+呋喃唑酮226.113915+226.1122*20+氯霉素3212574-321152*12-氟苯尼考356336*4-35618514-孔雀石绿329313.2*36+329208.132+孔雀石绿-D5334318.1*38+隐色孔雀石绿331315.228+331239.2*28+隐色孔雀石绿-D6337240.1*30+环丙沙星332314.2*17+332288.213+二氟沙星400356.1*17+400299.129+依诺沙星321.4303.3*19+321.4233.922+恩诺沙星360342.217+360316.2*17+氟甲喹262.3244.2*16+262.3202.136+洛美沙星351.933430+351.9265*36+诺氟沙星320302.2*17+320276.213+氧氟沙星362.4318.2*17+362.4261.125+培氟沙星334316.2*17+334290.213+沙拉沙星386342.213+386299.2*25+噁喹酸262.2244*12+262.221628+磺胺苯酰276.9156*12+276.910828+乙酰磺胺215156*8+21510812+磺胺氯哒嗪285156*12+28510820+磺胺嘧啶251156*12+25110820+磺胺甲噁唑254156.19+254108.1*21+磺胺间二甲氧嘧啶311156*16+31110828+磺胺邻二甲氧嘧啶311156*16+31110824+磺胺甲嘧啶265156*15+26510828+磺胺二甲嘧啶27918613+279124.1*25+磺胺甲噻二唑271156*12+27110816+磺胺苯吡唑31522220+315158*24+磺胺吡啶250156*16+25010824+磺胺噻唑256156*12+25610824+磺胺二甲异噁唑268156*12+268112.716+金霉素47946215+479444*22+多西环素445428*15+44515415+四环素44542725+445410*20+阿奇霉素749.5591.427+749.5158*36+克拉霉素748.5590.118+748.5158.1*28+红霉素734.4576.218+734.4158.2*28+交沙霉素828.3174.129+828.3109*40+吉他霉素772.4174.329+772.4109*40+替米考星869.569636+869.5174*48+泰乐菌素916.4174.2*37+916.4772.225+*为定量离子。线性范围在选定的仪器工作条件下，以各目标药物的特征离子的峰面积（响应值）为纵坐标，以为各目标药物的浓度为横坐标，绘制标准曲线。由于各类兽药物质在液相色谱-串联质谱仪器上的响应值不同，故根据响应值大小，将7类兽药物质分成两组浓度，建立一条由两个浓度系列组成的标准曲线，即硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、大环内酯类浓度为0.5ng/mL、1ng/mL、2ng/mL、5ng/mL、10ng/mL，四环素类浓度为5ng/mL、10ng/mL、20ng/mL、50ng/mL、100ng/mL，并最终含1ng/mL同位素内标物（氘代孔雀石绿、氘代隐性孔雀石绿）的标准曲线。结果表明，硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、大环内酯浓度在0.5ng/mL~10ng/mL，四环素类浓度在5ng/mL~100ng/mL，各种药物的浓度与响应值具有良好的相关性，各相关系数R>0.99，详见表3。保留时间与线性汇总表保留时间（min）线性相关系数r4.661y=3077.472792x-253.725982r=0.9976.865y=2920.813587x-123.364344r=0.9967.104y=2111.791346x86.004207r=0.9987.044y=8299.103293x-246.884952r=0.9997.233y=1.567939x-0.028941r=0.9999.984y=1.023069x-0.055921r=0.9994.917y=7751.071543x-1431.335655r=0.9965.765y=9595.421883x-2068.825708r=0.9964.701y=17671.83483x-3468.002853r=0.9985.212y=15308.715468x-5235.501012r=0.9977.832y=117424.797445x-3834.332653r=0.9995.074y=9381.764916x-766.955054r=0.9994.800y=7308.815379x-1291.903384r=0.9984.836y=28035.357253x-3366.366036r=0.9984.877y=7535.130526x-870.717865r=0.9975.666y=6991.751259x-670.253443r=0.9997.179y=149425.795711x-391.376508r=0.9987.316y=10124.81345x-441.121977r=0.9995.143y=6413.579497x-480.937027r=0.9986.900y=4096.417382x+120.419946r=0.9985.380y=3662.993981x+109.287917r=0.9997.022y=3647.64154x307.139741r=0.9986.998y=28035.438512x-1724.747113r=0.9997.334y=17169.444386x-2442.135847r=0.9975.971y=3812.841688x-122.442869r=0.9984.604y=22679.409322x-1100.460791r=0.9996.307y=4526.088191x-314.44665r=0.9977.373y=8360.187454x-343.749898r=0.9995.637y=4776.75244x+10.12838r=0.9995.359y=6901.824674x-139.639756r=0.9997.119y=5693.445727x-729.508177r=0.9975.803y=1425.152673x-123.345843r=0.9996.457y=6736.851749x-804.813481r=0.9974.992y=5268.053472x-1041.623093r=0.9976.910y=13474.528854x-1484.029963r=0.9986.910y=45362.79034x-5414.125109r=0.9986.673y=1442.965261x-89.0611r=0.9997.048y=23096.372957x-3258.18317r=0.9996.870y=11123.97815x-693.525122r=0.9996.275y=4421.565992x-19.053743r=0.9996.710y=7687.802923x+1050.344595r=0.997结果的计算和表述水样中硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、四环素类和大环内酯类药物的残留量按式（1）计算。

Xi=Ci×VV0式中：Xi——水样中待测药物残留量的含量，单位为微克每升(μg/L)；Ci——试样液中待测药物残留量的浓度，单位为微克每升(μg/L)；V——最终定容体积，单位为毫升(mL)；V0——测定水样品的体积，单位为毫升(mL)。方法检出限及定量限的确定本标准采用3倍信噪比计算仪器的检出限，将仪器检出限带入结果公式计算得到方法检出限详见表4，采用2倍检出限计算方法的定量限。经综合评定，取硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、大环内酯类的检出限为0.01μg/mL，定量限为0.02μg/mL；四环素类的检出限为0.10μg/mL，定量限为0.20μg/mL。方法检出限计算结果表药物名称信噪比测试浓度μg/L计算方法检出限μg/L呋喃它酮30.70.50.0010呋喃唑酮76.00.50.0004氯霉素487.20.50.0001氟苯尼考77.50.50.0004孔雀石绿444.90.50.0001隐色孔雀石绿318.60.50.0001环丙沙星16.60.50.0019二氟沙星43.90.50.0007依诺沙星51.50.50.0006恩诺沙星51.20.50.0006氟甲喹78.40.50.0004洛美沙星49.60.50.0007诺氟沙星28.80.50.0011氧氟沙星136.90.50.0003培氟沙星29.30.50.0011沙拉沙星75.30.50.0004噁喹酸56.20.50.0006磺胺苯酰/苯甲酰磺胺46.40.50.0007乙酰磺胺85.60.50.0004磺胺氯哒嗪34.10.50.0009磺胺嘧啶713.30.50.0001磺胺甲噁唑389.90.50.0001磺胺间二甲氧嘧啶112.60.50.0003磺胺邻二甲氧嘧啶107.90.50.0003磺胺甲嘧啶69.20.50.0005磺胺二甲嘧啶1566.40.50.0001磺胺甲噻二唑23.50.50.0013磺胺苯吡唑680.50.0005磺胺吡啶90.90.50.0004磺胺噻唑65.50.50.0005磺胺二甲异恶唑56.40.50.0006金霉素15.550.0194多西环素31.550.0096四环素1350.0231阿奇霉素91.70.50.0004克拉霉素703.50.50.0001红霉素186.20.50.0002交沙霉素192.90.50.0002吉他霉素72.10.50.0005替米考星37.40.50.0009泰乐菌素136.20.50.0003回收率和精密度按试验方法，将3个梯度浓度的混合标准溶液加入空白养殖淡水和空白养殖海水样品中进行回收试验和精密度实验，每个加标做7个平行，计算回收率和精密度。硝基呋喃类、氯霉素类、三苯甲烷类、喹诺酮类、磺胺类、大环内酯类标准物质加入水平为0.5ng/mL、1.0ng/mL、5.0ng/mL；四环素类标准物质加入水平为5ng/mL、10ng/mL、50ng/mL。得到回收率均在60%~120%之间，RSD≦20%，满足检测要求。结果见表5、表6。淡水养殖用水回收率和精密度实验结果药物名称加入量ng测得量ng回收率%RSD%加入量ng测得量ng回收率%RSD%加入量ng测得量ng回收率%RSD%呋喃它酮0.50.4~0.5780.94~113.2810.2910.69~1.0369.2~102.9712.1253.72~4.1474.37~82.724.12呋喃唑酮0.50.37~0.5674.81~111.1914.7910.64~0.8864.13~88.079.6553.58~4.1771.51~83.435.33氯霉素0.50.36~0.5171.33~101.1113.3810.6~0.9160.04~91.1113.2253.9~4.2378.09~84.63.39氟苯尼考0.50.36~0.4972.86~98.2410.2910.7~0.9670.31~96.1310.4953.82~4.476.38~87.995.09孔雀石绿0.50.46~0.5191.29~101.114.0110.83~0.8782.52~87.162.2953.84~4.1176.7~82.292.27隐色孔雀石绿0.50.35~0.3970.84~77.782.9810.69~0.7368.88~73.152.2054.2~4.3283.9~86.451.05环丙沙星0.50.49~0.5998.08~117.766.4610.74~0.8874.22~87.835.9955.45~5.78108.99~115.572.24二氟沙星0.50.47~0.5594.45~109.956.8410.67~0.8466.77~83.578.1854.92~5.3798.49~107.363.22依诺沙星0.50.47~0.5494.27~107.624.2410.69~0.7968.65~78.594.5155.46~5.89109.27~117.92.59恩诺沙星0.50.42~0.5183.08~102.667.2610.61~0.6961.17~69.384.7855.73~5.98114.53~119.551.44氟甲喹0.50.4~0.4580.73~90.994.2510.74~0.874.07~80.022.7354.85~5.0896.91~101.61.75洛美沙星0.50.52~0.57103.89~1132.6310.79~0.8479.33~83.651.9454.89~5.1997.73~103.891.98诺氟沙星0.50.49~0.5798.84~114.195.3710.78~0.8778.16~86.844.0655.58~5.91111.58~118.272.24氧氟沙星0.50.38~0.4476.93~87.534.5210.6~0.6960.11~69.234.1655.69~5.96113.88~119.121.68培氟沙星0.50.38~0.4976.44~98.159.9010.61~0.6860.74~67.713.9255.82~5.95116.42~119.070.78沙拉沙星0.50.46~0.5491.9~107.396.5410.75~0.8975.42~89.425.2854.68~5.0893.61~101.513.10噁喹酸0.50.58~0.6115.78~119.731.2110.92~0.9691.97~95.71.5654.92~5.1598.46~103.091.92磺胺苯酰/苯甲酰磺胺0.50.4~0.4580.95~90.013.7710.68~0.8168.07~81.196.0853.68~3.9173.66~78.252.02乙酰磺胺0.50.47~0.5794.25~114.36.3410.78~0.977.94~90.345.1854.03~4.3780.69~87.362.68磺胺氯哒嗪0.50.36~0.4672.76~92.987.9010.72~0.9272.12~91.778.8453.83~4.1476.53~82.892.67磺胺嘧啶0.50.41~0.5882.36~116.7811.1610.7~0.9369.81~92.669.2653.57~4.0671.32~81.244.12磺胺甲噁唑0.50.4~0.5279.55~103.158.6910.76~0.9375.94~93.378.5253.73~3.9974.59~79.762.71磺胺间二甲氧嘧啶0.50.34~0.5268.53~104.0314.8410.75~0.975.29~90.086.7154.09~4.3881.88~87.572.58磺胺邻二甲氧嘧啶0.50.38~0.4276.28~84.794.6410.74~0.8573.78~84.754.4454.11~4.2382.29~84.541.03磺胺甲嘧啶0.50.4~0.4479.99~88.653.6510.76~0.8476.13~83.843.9553.87~4.1577.35~82.932.29磺胺二甲嘧啶0.50.44~0.5687.08~112.778.3010.62~0.9361.87~92.8712.6253.84~4.1376.83~82.583.02磺胺甲噻二唑0.50.48~0.5295.33~104.23.4010.88~0.9387.56~92.642.1253.97~4.1479.45~82.711.64磺胺苯吡唑0.50.5~0.699.39~119.597.3410.91~1.0691.49~106.035.0354.23~4.684.57~92.093.340.50.45~0.5190.64~101.354.2510.77~0.9177.11~90.575.6953.61~4.1572.12~83.065.130.50.33~0.4465.62~87.8410.3710.73~0.8872.54~88.257.7353.91~4.2278.15~84.393.06磺胺二甲异恶唑0.50.37~0.5474.94~108.0512.6510.73~0.8773.35~86.935.4453.7~4.0374.08~80.523.38金霉素50.39~0.5478.79~107.6910.49100.64~0.8363.83~82.79.44503.96~4.9379.1~98.628.40多西环素50.39~0.5877.21~115.611.96100.64~0.6964.25~68.892.77505.48~5.96109.58~119.253.03四环素50.45~0.5289.53~103.965.59100.53~0.6553.42~65.366.94504.26~4.4485.23~88.721.74阿奇霉素0.50.4~0.4679.32~91.245.0710.65~0.7765.15~77.355.5953.02~3.2760.47~65.462.86克拉霉素0.50.39~0.4477.72~88.434.2410.7~0.7870.07~78.494.0453~3.1560.06~63.11.86红霉素0.50.43~0.5586.86~109.19.1810.76~1.0976.01~108.7612.3453.72~4.674.31~92.027.20交沙霉素0.50.45~0.4890.3~96.82.4510.8~0.8679.86~86.32.5653.91~4.1678.28~83.192.15吉他霉素0.50.48~0.5995.98~117.557.3610.83~0.9583.47~95.334.5654.1~4.3881.95~87.662.32替米考星0.50.47~0.5794.4~114.757.4810.82~0.9582.3~95.35.8654.16~4.8583.23~96.925.58泰乐菌素0.50.43~0.4785.15~93.934.0110.7~0.7970.28~78.74.6653.43~3.6268.55~72.392.06海水养殖用水回收率和精密度实验结果药物名称加入量ng测得量ng回收率%RSD%加入量ng测得量ng回收率%RSD%加入量ng测得量ng回收率%RSD%呋喃它酮0.50.43~0.5485.86~108.838.8710.79~0.9478.67~97.528.2453.66~4.3973.26~87.745.60呋喃唑酮0.50.36~0.5371.4~106.9812.2810.75~1.0274.67~102.0510.1553.69~4.273.86~83.944.42氯霉素0.50.33~0.5466.99~108.2216.6310.64~0.9863.61~98.3718.7853.52~4.5270.49~90.348.56氟苯尼考0.50.38~0.4576.68~90.697.0610.69~0.977.56~90.095.7253.85~4.4977.01~89.835.33孔雀石绿0.50.49~0.5397.27~105.632.8810.89~0.9689.03~96.872.9854.12~4.3782.34~87.422.11隐色孔雀石绿0.50.46~0.4891.34~96.812.1810.77~0.876.74~80.181.4653.69~3.8773.71~77.421.64环丙沙星0.50.4~0.5179.72~102.867.8210.76~0.9675.97~89.386.6753.15~3.5863~71.625.31二氟沙星0.50.46~0.5291.37~103.734.4910.75~0.975.36~92.016.5253.18~3.5363.6~70.573.97依诺沙星0.50.42~0.5284.44~103.826.2610.81~0.9281.43~91.725.0553.28~3.4965.63~69.822.22恩诺沙星0.50.52~0.58103.47~115.373.5310.81~0.9281.29~93.174.2853.07~3.1761.32~63.461.25氟甲喹0.50.39~0.4378.64~85.73.3710.77~0.8477.36~82.742.6453.72~4.0274.45~80.322.89洛美沙星0.50.34~0.4168.42~82.026.5210.73~0.8573.37~84.65.2453.31~3.7266.12~74.443.83诺氟沙星0.50.44~0.5188.44~102.515.7810.81~181.46~99.827.2453.21~3.6764.24~73.485.51氧氟沙星0.50.39~0.4378.53~86.823.5410.77~0.8777.15~87.463.9153.34~3.4866.72~69.541.45培氟沙星0.50.34~0.4168.39~82.667.7510.71~0.8670.75~85.536.0453.08~3.4561.62~68.923.83沙拉沙星0.50.36~0.4872.21~95.829.4610.77~0.8578.99~87.63.4553.62~3.7672.36~75.191.38噁喹酸0.50.36~0.3971.41~77.22.9910.76~0.8375.82~80.442.3453.69~3.8673.78~77.221.95磺胺苯酰/苯甲酰磺胺0.50.41~0.4481.43~87.512.9310.73~0.8773.12~86.715.9653.78~3.9575.65~78.931.67乙酰磺胺0.50.43~0.586.04~100.235.6010.71~0.9170.76~91.358.0553.97~4.3379.46~86.53.37磺胺氯哒嗪0.50.32~0.4964.51~97.2715.1610.61~0.8460.87~84.0610.5953.89~4.3177.87~86.184.00磺胺嘧啶0.50.32~0.4764.31~93.3713.8010.66~0.8265.82~82.096.6953.86~4.2477.11~84.824.09磺胺甲噁唑0.50.42~0.684.04~119.3511.4610.81~0.9674.25~95.778.0754.04~4.4380.81~88.563.08磺胺间二甲氧嘧啶0.50.43~0.5586.98~110.179.4010.79~0.9779.42~978.6753.76~4.2175.16~84.283.90磺胺邻二甲氧嘧啶0.50.43~0.4986.64~97.033.9610.79~0.8678.91~86.082.9153.94~4.2578.88~84.922.25磺胺甲嘧啶0.50.51~0.57101.37~113.094.1110.83~0.9183.22~90.763.0353.9~4.2278.09~84.332.55磺胺二甲嘧啶0.50.41~0.5181.32~102.088.8910.72~0.8773.73~86.666.6753.74~4.374.73~85.995.05磺胺甲噻二唑0.50.4~0.4779.93~94.016.3010.8~0.8376.82~82.912.4154.04~4.2480.83~84.91.78磺胺苯吡唑0.50.42~0.4683.38~91.933.6110.73~0.8673.47~85.545.2354.01~4.280.16~83.971.690.50.33~0.4865.82~95.0713.1810.68~0.8267.63~82.336.0353.75~4.0475.02~80.752.580.50.35~0.4769.26~93.8511.3910.72~0.8572.96~84.956.1253.97~4.1979.34~83.832.04磺胺二甲异恶唑0.50.46~0.5592.25~110.056.7710.77~0.976.55~89.766.0853.67~4.3473.43~86.75.48金霉素50.31~0.3961.55~77.567.61100.64~0.9363.87~93.0118.07503.03~3.260.61~63.952.09多西环素50.36~0.5172.98~101.9713.09100.79~0.9477.96~93.887.92503~3.1860.04~63.692.35四环素50.3~0.3860.51~75.857.71100.61~0.6460.53~64.392.25503.02~3.9460.41~78.8610.56阿奇霉素0.50.43~0.4886.42~96.853.8110.65~0.7265.4~72.13.3653.05~3.261.06~64.091.68克拉霉素0.50.44~0.4987.26~97.063.6510.66~0.7364.92~72.534.0453.02~3.260.47~63.941.94红霉素0.50.42~0.5783.48~114.3812.8910.65~0.9365.44~92.3612.5453.62~4.1572.45~82.945.02交沙霉素0.5