超高效液相色谱-串联质谱法测定鸡肉、鸡蛋中氟苯尼考和氟苯尼考胺残留

目的建立超高效液相色谱-串联质谱准确测定鸡肉、鸡蛋中氟苯尼考和氟苯尼考胺残留的分析方法。方法鸡肉和鸡蛋样品加入D5-氯霉素内标,经氨化乙酸乙酯混合溶液超声离心提取,提取液减压蒸馏浓缩后,C18固相萃取柱净化处理,采用超高效液相色谱-串联质谱法对氟苯尼考和氟苯尼考胺同时进行检测。结果氟苯尼考线性范围为0.2～20μg/L,检出限为1.0μg/kg,定量限为3.0μg/kg,方法回收率为88.0%～108.0%,相对标准偏差为4.7%～6.4%;氟苯尼考胺线性范围为0.2～20μg/L,检出限为1.0μg/kg,定量限为3.0μg/kg,方法回收率为76.0%～93.1%,相对标准偏差为4.1%～7.2%。结论本方法精确、重现性好,适用于鸡肉、鸡蛋中氟苯尼考和氟苯尼考胺残留量的测定。     

UPLC-MS/MS法测定禽蛋中氟苯尼考及氟苯尼考胺

建立一种超高效液相色谱-串联质谱法同时测定禽蛋中氟苯尼考(FF)和氟苯尼考胺(FFA)含量的方法。样品经2%氨水-乙酸乙酯提取,PRiME HLB固相萃取(SPE)柱净化。采用Hypersil GOLD C18色谱柱(100 mm×2.1mm,1.9μm),以乙腈(B1)-0.1%甲酸水(A2)为流动相进行梯度洗脱。氟苯尼考和氘代氟苯尼考采用电喷雾负离子源,氟苯尼考胺采用电喷雾正离子源,多反应监测模式,以氘代氟苯尼考(氟苯尼考-d3)为氟苯尼考进行内标定量,氟苯尼考胺外标法定量。结果表明,氟苯尼考和氟苯尼考胺在0.2~10μg/L范围内均呈良好线性关系(r≥0.9978)。加标浓度分别为0.1,0.2和1μg/kg及0.5,1.0和5.0μg/kg,回收率分别在91.0%~100.9%和73.7%~84.3%之间,相对标准偏差均<3.52%(n=6)。氟苯尼考的检出限和定量限分别为0.05μg/kg和0.1μg/kg,氟苯尼考胺的检出限和定量限分别为0.25μg/kg和0.5μg/kg。该方法准确、灵敏、快速,能有效并同时检测禽蛋中氟苯尼考和氟苯尼考胺含量。     

气相色谱质谱法检测鸡蛋中氟苯尼考及氟苯尼考胺

建立了气相色谱飞行时间质谱法同时检测鸡蛋中氟苯尼考和氟苯尼考胺的残留。样品经氨化乙酸乙酯提取,采用气相色谱飞行时间质谱法测定。试验结果表明:在2~100 ng/mL的范围内,线性良好,氟苯尼考R²=0.999,氟苯尼考胺R²=0.998;该方法两种物质的检出限为0.5μg/kg;在0.5、1.5、10.0μg/kg的3个水平,氟苯尼考及氟苯尼考胺的回收率在86.4%~119.4%,相对标准偏差RSD(n=3)为2.9%~7.1%,该方法简单、快速、高效,可用于鸡蛋中氟苯尼考及氟苯尼考胺的同时测定。     

内标液相色谱-串联质谱法测定蜂蜜中氯霉素、甲砜霉素和氟苯尼考残留

目的建立液相色谱-串联质谱法测定蜂蜜中氯霉素、甲砜霉素和氟苯尼考的分析方法。方法以氯霉素-D5为内标,样品用乙酸乙酯提取,提取液在45℃氮气吹至近干,残渣用2mL水溶解,溶解液过Oasis HLB柱净化,洗脱液在45℃氮气吹干, 50%甲醇水溶液定容,后经正己烷脱色脱脂、PSA粉末吸附净化,高速离心过膜后用串联四极杆检测。样液经过Agilent Eclipse XDB-C₁₈色谱柱分离,甲醇-水流动相梯度洗脱,多反应监测扫描模式检测。采用内标法定量。结果氯霉素在0.1~4.0μg/kg、甲砜霉素和氟苯尼考在0~40.0μg/kg呈良好的线性关系,相关系数大于0.999,回收率为93.7%~109.7%,RSD为0.7%~11.4%,检出限为0.03~0.09μg/kg,定量限为0.1~0.3μg/kg。结论该方法准确、检出限低、重现性好,适用于蜂蜜中氯霉素、甲砜霉素和氟苯尼考残留的检测。     

液相色谱-串联质谱法快速测定鸡蛋中金刚烷胺、氟苯尼考和氟苯尼考胺残留量

目的建立同时测定鸡蛋中金刚烷胺、氟苯尼考及氟苯尼考代谢物氟苯尼考胺残留的液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry, LC-MS/MS)快速分析方法。方法鸡蛋样品用1.0%乙酸乙腈提取,提取后用PSA和C_(18)净化,氮吹后用流动相复溶,复溶液过滤膜后进行液相色谱-串联质谱测定。分析采用XBridge C_(18)色谱柱,以0.02%甲酸溶液和甲醇乙腈混合溶剂为流动相进行梯度洗脱,正负离子切换监测模式下进行监测,同位素内标法定量。结果3种化合物在一定浓度(氟苯尼考为0.10～20μg/L,金刚烷胺和氟苯尼考胺为0.5～100μg/L)范围内具有较好的线性关系,金刚烷胺、氟苯尼考和氟苯尼考胺的检出限(limit of detection, LOD)分别为0.3、0.06、0.3μg/kg,定量限(limit of quantitation, LOQ)分别为1.0、0.20、1.0μg/kg。当3种化合物在鸡蛋中的加标水平为LOQ、5LOQ和10LOQ时,平均回收率在94.9%～102%之间,相对标准偏差范围为1.2%～4.9%。结论方法简便、快速、准确,能满足鸡蛋中金刚烷胺和氟苯尼考残留量同时分析需要。     

Captiva EMR-Lipid固相萃取结合高效液相色谱-串联质谱法同时测定猪肉中氯霉素类药物残留量

目的 建立Captiva EMR-Lipid固相萃取结合高效液相色谱-串联质谱法同时测定猪肉中氯霉素、甲砜霉素和氟苯尼考残留量的分析方法。方法 采用乙腈提取样品, 经Captiva EMR-Lipid固相萃取柱净化, 以甲醇-水为流动相, CNW Athena C18-WP色谱柱(100 mm×2.1 mm, 3 μm)进行分离, 采用电喷雾负离子扫描和多反应监测模式, 内标法定量。结果 氯霉素、甲砜霉素和氟苯尼考在0.5~20.0 ng/mL范围内呈现良好的线性关系(r＞0.9975), 方法的检出限为0.10~0.14 ?g/kg, 定量限为0.25~0.47 ?g/kg, 平均回收率为97.50%~117.00%, 相对标准偏差为5.55%~8.93%。结论 该方法简单快速、稳定可靠, 适用于猪肉中氯霉素、甲砜霉素和氟苯尼考的同时测定。     

QuEChERS-高效液相色谱-串联质谱法同时测定猪肉中氯霉素、氟苯尼考和五氯酚的残留量

目的建立QuEChERS-高效液相色谱-串联质谱法快速测定猪肉中氯霉素、氟苯尼考和五氯酚的分析方法。方法以乙腈为提取溶剂,加入正己烷除脂,氮吹浓缩乙腈定容后经C_(18)填料萃取净化,在高效液相色谱-串联质谱仪上采用电喷雾负离子扫描模式(electron spray ionization,ESI-)测定,内标法定量。结果该方法在1.0～50.0μg/L范围内有良好的线性关系,相关系数均大于0.99,氯霉素和氟苯尼考的方法检出限为0.1μg/kg,五氯酚的方法检出限为0.06μg/kg。平均回收率在81.1%～115.8%范围,相对标准偏差在4.2%～11.8%范围。结论本方法操作简便,准确度和精密度好,可同时检测猪肉中氯霉素、氟苯尼考和五氯酚的残留量。     

超高效液相色谱串联质谱法测定液态奶中氯霉素、甲砜霉素和氟苯尼考含量

建立超高效液相色谱串联质谱法测定液态奶中氯霉素、甲砜霉素和氟苯尼考的检测方法。样品经乙酸乙酯提取,Oasis MCX小柱净化,用色谱柱WATERS ACQUITY UPLC BEH C18(1.7um, 2.1mm*100mm)分离,同位素内标法定量。在优化条件下,该方法中氯霉素的最低检出限为0.05μg /kg,定量限为0.15μg /kg, 甲砜霉素和氟苯尼考的最低检出限为0.1μg /kg,定量限为0.3μg /kg, 平均回收率为80.4~105.4%,相对标准偏差为2.8~8.4%。该方法灵敏度高、回收率好,具有良好的净化效果,提升检测效率,适用于液态奶中氯霉素、甲砜霉素和氟苯尼考的同时检测。     

高效液相色谱-串联质谱法同时检测虾肉和猪肉中氯霉素、甲砜霉素、氟苯尼考和其代谢产物氟苯尼考胺残留

建立了高效液相色谱-串联质谱法同时检测虾肉和猪肉中氯霉素、甲砜霉素、氟苯尼考和其代谢产物氟苯尼考胺残留。以2%碱性乙酸乙酯提取,省去固相萃取(SPE)和液-液萃取(LPE)脱脂,浓缩后采用甲醇和5mmol/L乙酸铵以0.25 mL/min流速梯度洗脱,LC-MS/MS电喷雾电源(ESI),负离子,选择反应监测(SRM)模式检测。试验结果表明:4种药物在0.1~20μg/L的范围内线性良好,相关系数R2在0.9997~0.9998之间。在添加量为1.00~5.00μg/kg时,平均回收率在78.17%~99.86%之间,RSD在2.18%~9.27%之间。氯霉素、甲砜霉素、氟苯尼考和氟苯尼考胺的检出限分别为0.001,0.020,0.002,0.003μg/kg。     

固相萃取-高效液相色谱-串联质谱法同时测定蜂蜡中氯霉素、甲砜氯霉素、氟苯尼考和其代谢产物氟苯尼考胺残留

首次建立了一种同时测定蜂蜡中氯霉素、甲砜氯霉素、氟苯尼考和其代谢物氟苯尼考胺等药物残留的固相萃取-高效液相色谱-串联质谱分析方法。根据蜂蜡脂溶性的特点,试样采用正己烷溶剂预溶解,用水提取2次后经混合型阳离子交换固相萃取柱（MCX）净化,用InertSustain Swift C₁₈柱（100 mm×2.1 mm, 1.9 μm）分离,多反应监测（MRM）模式串联质谱进行测定,以内标法定量。结果表明,在0.1~10 ng/mL范围内4种测定物均具有较好的线性关系,相关系数大于0.99,方法定量限（S/N>10）为0.1 μg/kg。对阴性蜂蜡在0.1、0.3和1.0 μg/kg等3个添加水平下分别进行加标回收试验,实际样品中4种测定物质的平均回收率在65.0%~113.2%之间,相对标准偏差（RSD,n=6）小于20%。应用本方法对15个蜂蜡样品进行检测,结果表明有4批次样品检出氯霉素类药物残留。本方法快速、灵敏、准确,适用于日常大批量蜂蜡样品中氯霉素类药物及其代谢物残留的定性、定量分析。     

氟苯尼考及其代谢物氟苯尼考胺在肉鸡中残留消除规律

目的 评估氟苯尼考及其代谢物氟苯尼考胺在不同品种肉鸡中的残留消除规律。方法 选用AA肉鸡、WOD168肉鸡、雪山鸡和狼山鸡为试验对象, 于出栏前3周开始按氟苯尼考25 mg/kg体重饮水给药, 连用5 d, 于停药后1、3、5、7、10、14 d采集肌肉、肝脏、肾脏、皮脂, 检测其中的氟苯尼考和氟苯尼考胺残留量, 并利用WT1.4软件计算休药期。结果 氟苯尼考及其代谢物氟苯尼考胺可迅速残留于肉鸡肌肉、肝脏、肾脏和皮脂中, 残留物以氟苯尼考为主。停药后第1 d残留量达到峰值, 肾脏组织中的残留量最大; 停药后第3 d, 各组织中氟苯尼考和氟苯尼考胺的残留量进入快速消除期, 均低于最大残留限量值; 停药后第14 d已基本消除完全。利用WT1.4软件计算出AA肉鸡、WOD168肉鸡、雪山鸡和狼山鸡休药期分别至少为3.22、3.85、4.49和4.32 d。结论 氟苯尼考在慢速型肉鸡体内的休药期要长于快速型肉鸡, 但都小于5 d。     

Simultaneous determination of chloramphenicol,florfenicol and florfenicol amine in ham sausage with a hybrid chemiluminescent immunoassay

A novel chemiluminescent immunoassay utilising two types of primary antibodies (murine monoclonal antibody and rabbit polyclonal antibody) and two types of horseradish peroxidase–labelled secondary antibodies was established for simultaneously detecting multiple amphenicol residues in ham sausage. After combining the extract procedure of the target amphenicol into one simplified method, this hybrid chemiluminescent immunoassay could screen chloramphenicol (CAP), florfenicol (FF) and its metabolite florfenicol amine (FFA) at the same time by adding the corresponding secondary antibody. Ham sausage samples were analysed by using this hybrid immunoassay, with LODs of CAP being 0.01 μg kg^?1, of FF being 2.8 μg kg^?1 and of FFA being 3.0 μg kg^?1. The applicability of the proposed method has been validated by determining CAP, FF and FFA in ham sausage samples with satisfactory results. Good recoveries and high correlation with traditional enzyme-linked immunosorbent assay and LC-MS/MS results illustrated that the developed hybrid chemiluminescent immunoassay could screen high-throughput ultra-trace amphenicol residues effectively at one time.     

A reliable,simple and cost-efficient TLC-HPLC method for simultaneously determining florfenicol and florfenicol amine in porcine urine: application to residue surveillance

ABSTRACT

Violative residues of florfenicol (FF) in porcine edible tissues pose a potential risk for human health. In this study, urine was selected as target matrix for routine residue monitoring of FF in pig, and a thin layer chromatography (TLC)-high-performance liquid chromatography (HPLC) method was developed for simultaneously determining FF and florfenicol amine (FFA) in porcine urine. The urine samples were extracted with ethyl acetate under alkaline environment. The extracts were enriched through evaporation, purified by TLC and analysed by HPLC at 225 nm. A Waters Symmetry C₁₈ column was used for the separation of the two analytes. The mobile phase was acetonitrile-phosphate buffer mixtures (33.3: 66.7, v/v), and was pumped at 0.6 mL/min. The TLC-HPLC method was well validated and successfully applied to residue depletion study. Good analytical specificity was confirmed by the lack of interfering peaks at the retention times of FF and FFA. The standard curves showed good linearity (FF: y = 143064x – 1045.3, r= 0.9999; FFA: y = 275826x + 1888.8, r= 0.9999) over the range of 0.0625–8 μg/mL. The precision ranged from 0.83% to 11.66% and 2.19% to 8.75% for intraday and interday determination, respectively. The corresponding accuracy ranged from ?13.38% to 10.78% and ?12.15% to 7.14%, respectively. The limits of quantification (LOQs) for FF and FFA were 0.125 μg/mL. The residue depletion study showed that the concentrations of FF and FFA in urine were higher than those in edible tissues at three time points. This method was reliable, simple and cost efficient, and could be used to monitor FF residues in porcine edible tissue without slaughtering animals. TLC showed excellent purification efficiency and is expected to solve matrix interferences in veterinary drug residue analysis.     

LC-MS/MS-based determination of chloramphenicol,thiamphenicol, florfenicol and florfenicol amine in poultry meat from the Punjab-Pakistan

A simple, reliable and sensitive liquid chromatography-tandem mass spectrometry-based confirmatory method was redeveloped and validated for the simultaneous determination of chloramphenicol, thiamphenicol, florfenicol and florfenicol amine in chicken muscles. The analytes were extracted from minced chicken muscle with acetonitrile and ammoniated water mixture. A second extraction with ethyl acetate was followed by evaporation and dissolution of the residue in ammoniated methanol before defatting with n-hexane. Finally, the extract was further cleaned up by dispersive solid phase extraction using C-18 end-capped dispersive material. The validation protocol was adapted from the European Commission Decision 2002/657/EC and all the performance characteristics were successfully satisfied. The recoveries of all the analytes were found to be in the range of 86.4–108.1% and the precision values, within day and between days, ranged from 2.7% to 11% and 4.4% to 16.3%, respectively. The method was tested in various incurred samples and applied to analyse a wide range of random poultry market samples (n = 120) collected from three cities of the Punjab, Pakistan. Chloramphenicol and florfenicol residues were detected at low levels in less than 11% of the samples. Chloramphenicol was detected only in 4 samples with the concentration range of 0.17–0.477 µg kg^–1, whereas the levels of florfenicol/florfenicol amine residues detected in 9 samples ranged from 8.7 to 32.8 µg kg^–1. Moreover, most of the florfenicol residues were identified as tissue bound, extractable only after strong acid hydrolysis.     

池塘养殖模式下氟苯尼考及其代谢产物在斑点叉尾鮰体内的药代动力学

目的在实际大池塘养殖模式下,研究氟苯尼考(florfenicol,FF)及其代谢产物在斑点叉尾鮰(Letaurus以下简称鮰)体内药代动力学。方法选择4口标准化池塘(2300 m~2/口),养殖密度按每666.67 m~2投放鱼苗1000尾投放,设3个实验组和1个对照组。实验组分别以1.25、2.50、5.00 g/kg 3个剂量水平在鮰饲料中添加FF,每天投喂饲料4 kg(一次投入),对照组投喂不含FF的等量饲料。连续投喂5 d后,分别于首次投药后的第1、2、3、4、5、6、7、8、11、14、30、60、90和170 d采集鮰肌肉、肝脏及血液,采用高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)检测FF及氟苯尼考胺(florfenicol amine,FFA)含量,采用PKSolver药动学药效学数据处理软件V2.0的药动学房室模型拟合方法分析药时数据。结果 FF在鮰血液中的药代动力学特征符合一级吸收一室模型,而在肝脏及肌肉中均符合一级吸收二室模型,FFA在不同组织中均符合一级吸收二室模型。结论 FF在鮰体内主要以原型的方式代谢消除,分布广泛且FF消除速率远高于FFA,肝脏可作为残留分析的靶组织。     

鸭蛋中的氟苯尼考药物残留的检测及调查研究

目的对氟苯尼考(florfenicol, FFC)在鸭蛋中的残留情况做初步的调查研究。方法针对浙江南部、福建北部、广东南部、湖北中部、江西东部、河南东部的六大区域的鸭蛋采用GB/T 22338-2008《动物源性食品中氯霉素类药物残留量测定》规定的方法进行氟苯尼考的药残检测。结果标准曲线相关系数为:r=0.9996,方法准确。以该方法测定每个地区6组鸭蛋样品,6组咸鸭蛋样品,共计12组样品,其中以河南东部出产的鸭蛋为原料制成的咸鸭蛋检出氟苯尼考残留量为:1.44μg/kg,其余样品未检出。结论氟苯尼考在鸭蛋中会造成一定程度的残留,仍需通过严格的养殖现场管理控制用药,并且通过鸭蛋的定期抽检来监测指标的变化。     

基于纳米金比色法可视化检测鸡蛋中的氟苯尼考

该研究基于适配体功能化的纳米金构建了比色传感器,在优化各种试验条件的基础上,评估了传感器的灵敏性及特异性,并对鸡蛋中氟苯尼考检测的可行性进行了研究,进而探索将比色传感器与智能手机的成像分析相结合实现快速分析的可行性.比色传感器的优化条件如下:NaCl浓度为50 nmol/L,NaCl孵育时间为5 min,适配体浓度为7...