兽药中非法添加药物快速筛查法参考模板范本

附件兽药中非法添加药物快速筛查法（液相色谱—二极管阵列法）1适用范围1.1本方法适用于兽药及其原料与辅料中紫外光谱图库中所列非法添加药物的筛查。1.2用于紫外光谱图库之外的具有紫外吸收的非法添加药物的筛查时，需进行空白试验和检测限测定，并测定对照品溶液的紫外光谱图。2检查方法照高效液相色谱法（《中国兽药典》一部附录0512）测定。2.1超高效液相色谱—二极管阵列法色谱条件与系统适用性试验用十八烷基硅烷键合硅胶为填充剂（ACQUITYUPLCHSST3，1.8µm，2.1×100mm）；以水-1mol/L醋酸铵溶液（取醋酸铵77g，加水至1000ml，用冰醋酸调节pH值至5.0）（99∶1）为流动相A，甲醇-1mol/L醋酸铵溶液（99∶1）为流动相B，按下表进行梯度洗脱；流速0.45ml/min；二极管阵列检测器，采集波长范围为200nm～400nm，分辨率为1.2nm，提取波长按需设定（如230nm、270nm、最大值图等）。时间（min）流动相A（%）流动相B（%）0.009820.2598212.2519913.0019913.0198217.00982供试品溶液的配制中药固体制剂取50mg，加甲醇10ml，超声处理10分钟（必要时可调整）；中药液体制剂取10µl~20µl，加甲醇10ml，摇匀；化药液体制剂取20µl，加甲醇10ml，摇匀；其他化药制剂取50mg，加甲醇10ml，超声处理约10分钟（必要时可调整），用甲醇稀释成含有效成分50µg~100µg/ml的溶液。对照品溶液的配制（紫外光谱图库外品种）用甲醇制成50~100µg/ml的溶液，溶解性差的对照品，可加适量盐酸或氢氧化钾溶液，使其溶解。测定法取供试品溶液2µl注入液相色谱仪，同时记录色谱图与光谱图。通过与紫外光谱图库中对照光谱图最大吸收波长和图形的比对，确定供试品中是否含有相应非法添加药物。可根据实验结果调整进样量或浓度，使紫外光谱图光滑、特征清晰。结果判定两者光谱图无明显差异，判为可疑阳性结果。筛查结果呈可疑阳性的样品，需按农业农村部发布的非法添加检查方法标准进一步检验。2.2液相色谱—二极管阵列法色谱条件与系统适用性试验用十八烷基硅烷键合硅胶为填充剂（AlltimaC18，5µm，4.6×250mm）；以磷酸二氢钠溶液（取磷酸二氢钠3.0g，加水1000ml，加三乙胺0.5ml，用饱和氢氧化钠溶液调pH值至7.0）∶甲醇（70∶30）为流动相；流速1.0ml/min；二极管阵列检测器，采集波长范围为200nm～400nm，分辨率为1.2nm，提取波长按需设定。供试品溶液的配制同2.1。对照品溶液的配制同2.1。测定法取供试品溶液10µl~20µl注入液相色谱仪，其他同2.1。结果判定同2.1。2.3液相色谱—二极管阵列法（磺胺类）色谱条件与系统适用性试验用十八烷基硅烷键合硅胶为填充剂（AlltimaT3，5µm，4.6×250mm）；0.2%冰醋酸溶液∶乙腈（70∶30）为流动相；流速1.0ml/min；二极管阵列检测器，采集波长范围为200nm～400nm，分辨率为1.2nm，提取波长按需设定。供试品溶液的配制同2.1。对照品溶液的配制同2.1。测定法取供试品溶液10µl~20µl注入液相色谱仪，其他同2.1。结果判定同2.1。2.4液相色谱—二极管阵列法（氟喹诺酮类）色谱条件与系统适用性试验用十八烷基硅烷键合硅胶为填充剂（AtlantisC18，5µm，4.6×250mm）；以磷酸溶液（取磷酸3ml，加水1000ml，用三乙胺调节pH值至3.0±0.1，加乙腈53ml，摇匀）为流动相A，乙腈为流动相B，甲醇为流动相C，按下表进行梯度洗脱；流速1.0ml/min；二极管阵列检测器，采集波长范围为200nm～400nm，分辨率为1.2nm，提取波长按需设定。时间（分钟）流动相A%流动相B%流动相C%089.66.24.230.0089.66.24.230.209010055.009010055.2089.66.24.265.0089.66.24.2供试品溶液的配制同2.1。对照品溶液的配制同2.1。测定法取供试品溶液10µl~20µl注入液相色谱仪，其他同2.1。结果判定同2.1。附：紫外光谱图库

紫外光谱图库名称编号最大吸收波长(nm)谱图氨茶碱001-1206.9273.5001-2201.5271.2茶碱002-1208.1273.5002-2201.5271.2注：编号由品种序号和方法序号组成，-1是指方法2.1，-2是指方法2.2，-3是指方法2.3，-4是指方法2.4，对应谱图为该方法条件下所得。二羟丙茶碱003-1208.1275.9003-2205.1272.4己酮可可碱004-1208.1275.9004-2206.2273.6烟酰胺005-1216.4264.0005-2213.3261.7喹乙醇006-1204.5241.3262.8006-2238.1260.5乙酰甲喹007-1242.5259.2007-2239.2255.8喹烯酮008-1234.2261.6316.7008-2不出峰卡巴氧009-1240.1308.3378.8009-2236.9304.5376.0诺氟沙星010-1209.3280.7317.9010-2228.6272.4323.6010-4278.5吡哌酸011-1218.8274.7326.3011-2226.3264.1331.9氧氟沙星012-1228.3296.3012-2226.3254.6287.8330.7012-4226.4293.9环丙沙星013-1288.7319.1013-2271.2323.6013-4278.5315.4洛美沙星014-1212.8289.1323.9014-2280.7326.0014-4288.0320.1达氟沙星015-1285.5347.5015-260分钟不出峰015-4283.2347.6沙拉沙星016-1281.9321.5016-2232.2273.6323.6016-4280.8316.6培氟沙星017-1280.7319.1017-260分钟不出峰017-4277.3315.4恩诺沙星018-1280.7319.1018-2120分钟以后出峰018-4277.3315.4二氟沙星019-1281.9321.5019-260分钟不出峰019-4279.7316.6磺胺嘧啶020-1268.8020-2257.0020-3267.8磺胺噻唑021-1260.4287.9021-2257.0279.5021-3258.3285.6磺胺二甲嘧啶022-1267.6022-3266.6磺胺对甲氧嘧啶023-1271.1023-2242.8磺胺氯哒嗪024-1270.0磺胺甲噁唑025-1270.0025-2257.0磺胺间甲氧嘧啶026-1273.5磺胺氯吡嗪027-1270.0027-2257.0330.7磺胺喹噁啉028-1250.9344.1028-2251.1360.0阿司匹林029-1202.2029-2195.7水杨酸030-1204.5231.8298.7030-2201.5229.8295.0水杨酸镁031-1204.5233.0298.7吲哚美辛032-1208.1322.7032-2201.5228.6264.1320.0水相:有机相40:60对乙酰氨基酚033-1203.4246.1033-2196.8244.0氨基比林034-1203.4233.0268.8034-2196.8228.6265.3380.8安替比林035-1203.4244.9261.6035-2196.8241.6安乃近036-1203.4231.8036-2198.0228.6MAA（安乃近降解产物）037-1201.0249.7酮洛芬038-1204.5260.4038-2199.2258.2水相:有机相40:60富马酸酮替芬039-1204.5301.1芬布芬040-1204.5286.7040-2199.2283.1水相:有机相40:60布洛芬041-1202.2223.5266.4041-2196.8221.5264.1水相:有机相40:60氟比洛芬042-1209.3249.7双氯芬酸钠043-1204.5283.1043-260分钟不出峰甲芬那酸044-1215.2289.1344.1044-2199.2286.6334.3水相:有机相40:60萘普生045-1233.0273.5333.5045-2231.0261.7330.7水相:有机相40:60美洛昔康046-1206.9273.5365.5046-2203.9271.2361.0水相:有机相40:60甲硝唑047-1203.4231.8321.5047-2199.2228.6317.6地美硝唑048-1203.4230.6321.5048-2228.6317.6替硝唑049-1203.4233.0319.1氟苯尼考050-1203.4225.9268.8050-2223.9265.3甲砜霉素051-1203.4227.1268.8051-2223.9265.3氯霉素052-1205.7279.5052-2不出峰头孢吡肟053-1204.5237.8259.2头孢丙烯054-1203.4231.8281.9头孢地尼055-1225.9289.1头孢地嗪056-1204.5235.4262.8头孢唑啉057-1203.4274.7头孢西丁058-1203.4237.8头孢替唑059-1204.5274.7头孢呋辛060-1203.4275.9头孢呋辛酯061-1203.4288.7头孢甲肟062-1203.4234.2头孢克洛063-1203.4266.4332.3头孢克肟064-1206.9234.2290.3头孢喹肟065-1204.5236.6268.8头孢拉定066-1203.4264.0344.1头孢硫脒067-1211.7334.7头孢美他酯068-1203.4236.6头孢美唑069-1203.4241.3274.7头孢孟多酯070-1204.5271.1348.7头孢尼西071-1203.7270.0头孢哌酮072-1202.2230.6270.0头孢匹胺073-1212.8275.9头孢羟氨苄074-1203.4231.8265.2341.8头孢噻吩075-1203.4239.0329.9头孢噻呋076-1204.5236.6268.8头孢噻肟077-1204.5237.8头孢他啶078-1203.4258.0头孢替安079-1203.4261.6二甲氧苄啶080-1204.5279.5080-2199.2284.3甲氧苄啶081-1205.7273.5081-2201.5285.4呋喃妥因082-1202.2267.6369.1082-2198.0274.8378.4呋喃唑酮083-1202.2261.6367.9083-2259.3363.0呋喃西林084-1201.0262.8377.6084-2194.5260.5376.0克伦特罗085-1211.7246.1299.9085-2209.8242.8296.1莱克多巴胺086-1224.7277.1086-2195.7221.5274.8（双头峰）特步他林087-1203.4279.5087-2199.2279.5硫酸沙丁胺醇088-1202.2227.1279.5盐酸氯丙那林089-1201.0264.0089-2210.9260.5328.3盐酸班布特罗090-1202.2266.4盐酸丙卡特罗091-1206.9235.4261.6298.7341.8尼美舒利092-1203.4302.3盐酸二氧丙嗪093-1228.3266.4292.7329.9093-2223.9268.8295.0333.1水相:有机相40:60氢氯噻嗪094-1227.1273.5319.1094-2223.9271.2316.4布美他尼095-1202.2223.5334.7095-2不出峰盐酸氨溴索096-1211.7248.5313.1096-2208.6245.1302.1盐酸溴己新097-1210.5250.9309.5097-2不出峰长春西汀98-1205.7228.3273.5316.798-2不出峰依他尼酸99-1205.7281.9盐酸尼卡地平100-1206.9239.0357.1100-2不出峰尼莫地平101-1204.5240.1359.5101-2不出峰盐酸氯丙嗪102-1204.5258.0309.5102-2不出峰异丙托溴铵103-1202.2260.4阿昔洛韦104-1203.4254.4恩替卡韦105-1203.4255.6泛昔洛韦106-1223.5310.7磷酸奥司他韦107-1203.4醋酸倍他米松108-1201.0242.5二丙酸倍他米松109-1241.3倍他米松110-1201.0242.5戊酸倍他米松111-1202.2241.3丙酸倍氯米松112-1241.3倍氯米松113-1203.4243.7丁酸氯倍他松114-1201.0239.0地塞米松115-1242.5醋酸地塞米松116-1201.0242.5地塞米松磷酸钠117-1201.0242.5117-2不出峰氟米松118-1241.3甲氧氯普胺119-1215.2277.1311.9119-2212.1273.6310.4吡喹酮120-1204.5266.4120-2198.0262.9盐酸利多卡因121-1202.2121-2198.0盐酸吗啉胍122-1203.4239.0122-2198.0236.9芬苯达唑123-1220.0297.5123-2不出峰盐酸氯苯胍124-1204.5320.3124-2不出峰三氯苯达唑125-1204.5223.5308.3125-2不出峰氟甲喹126-1236.6326.3126-2215.7247.5330.7丙磺舒127-