杭州市动物性食品中喹诺酮类抗生素残留水平及安全性评价

目的了解杭州市动物性食品中喹诺酮类抗生素残留水平,探讨其污染来源,评价其膳食安全性。方法 2017年5月-11月随机采集杭州市动物性食品,包括水产品(河虾、小龙虾)和畜禽类(鸡肉、鸡蛋和猪肉),采用超高效液相色谱-串联质谱法进行喹诺酮类抗生素(氧氟沙星、培氟沙星、诺氟沙星、洛美沙星、环丙沙星、达氟沙星、二氟沙星、恩诺沙星、氟甲喹、恶喹酸、沙拉沙星)检测。结果 88份动物性食品喹诺酮类抗生素残留总检出率为19.4%,其中7份鸡蛋样品超标,超标率为7.9%。17份阳性样品中恩诺沙星检出比例最高(68.4%),对其进行膳食摄入安全性评价,未超过每日安全摄入量。结论杭州市市售动物性食品中存在部分抗生素残留,需要加强对喹诺酮类抗生素,尤其是禁用抗生素的生产、销售和使用全过程的管理。     

超高效液相色谱-串联质谱法分析动物组织中7种喹诺酮类药物残留

目的:建立超高效液相色谱-串联质谱法(UPLC/MS/MS)测定动物性食品中残留的7种喹诺酮类药物:培氟沙星、恩诺沙星、沙拉沙星、环丙沙星、二氟沙星、单诺沙星、氧氟沙星。方法:样品经Mellvaine缓冲液超声提取,高速离心后,用HLB固相萃取拄净化后上UPLC/MS/MS,用多反应监测模式检测,外标法定量。结果:7种喹诺酮类药物在浓度1μg/L～100μg/L范围内线性关系良好,相关系数均达到r>0.9985。检出限0.1μg/kg～1μg/kg。结论:本方法简便、快速、定性准确,适合动物性食品中残留的7种喹诺酮类药物残留测定。     

高效液相色谱-串联质谱测定鸡肉中氟喹诺酮类药物残留的研究

目的:建立鸡肉中培氟沙星、单诺沙星、双氟沙星、氧氟沙星、诺氟沙星、沙拉沙星、环丙沙星、恩诺沙星和司帕沙星等9种氟喹诺酮类药物残留的高效液相色谱-串联质谱检测方法.方法:样品经磷酸盐缓冲液提取后,用Waters Oasis (R) MAX小柱进行净化、提取,Cloversil-C18柱(150mm×4.6 mm i.d,5μm)分离;柱温:30℃;流动相:甲醇/乙腈/0.2%甲酸(15/15/70,v/v/v);通过电喷雾电离离子化在多离子监测(MRM)模式下测定.结果:各氟喹诺酮在0.05～50.0 μg/kg范围内均呈良好的线性关系,相关系数>0.998,回收率在89.0%～110.0%之间,相对标准偏差(RSD)在2.8%～9.1%之间,检出限均为0.05 μg/ks.结论:所建方法简便、快速、干扰少、特异性强,是鸡肉中9种氟喹喏酮残留检测的理想方法.     

蜂蜜中4种氟喹诺酮残留的高效液相色谱-质谱测定

目的:建立蜂蜜中氧氟沙星、恩诺沙星、双氟沙星、沙拉沙星4种氟喹诺酮残留的高效液相色谱-质谱联用检测方法。方法:样品经磷酸盐缓冲液稀释、溶解后,用Waters Oasis MAX小柱进行净化、提取,Cloversil-C18柱(150 mm×4.6 mm×5μm)分离;流速:0.5 ml/min;柱温:30℃;流动相:乙腈/0.05%三氟乙酸(25/75,v/v);通过电喷雾电离离子化在选择离子监测(SIM)模式下测定,氧氟沙星、恩诺沙星、双氟沙星、沙拉沙星定量分析离子m/z:362.0[M+H]+、360.1[M+H]+、400.0[M+H]+和386.0[M+H]+,外标法定量。结果:在0.50~100.0μg/kg范围内4种目标化合物均呈良好的线性关系,平均回收率大于95.3%,RSD均小于3.5%,检出限为0.5μg/kg。结论:本法简便、灵敏、重现性好、特异性强,是蜂蜜中4种氟喹诺酮残留检测的有效方法。     

贵阳市场常见淡水鱼体内氟喹诺酮类抗生素残留调查

目的了解贵阳市市售淡水鱼体内中氟喹诺酮类抗生素(FQs)的残留情况。方法于2015年10—11月,在贵阳市3个水产批发市场(A、B、C区)采集鲤鱼(Cyprinus carpio)、鮰鱼(Leiocassis longirostris)、黄搡鱼(Pelteobagrus fulvidraco)、黄蜂鱼(Yellow catfish)、鲈鱼(Lateolabrax japonicus)5种淡水养殖鱼样品,采用超高效液相色谱-串联质谱(UPLC-MS/MS)法检测鱼肌肉组织中7种FQs(恩诺沙星、双氟沙星、诺氟沙星、氧氟沙星、环丙沙星、丹诺沙星、依诺沙星)的残留量。结果淡水鱼体内恩诺沙星和氧氟沙星检出率较高(分别为92.9%和78.6%),平均含量分别为96.5、135.1μg/kg,最高分别为636.2、488.2μg/kg,恩诺沙星超过最大允许限量(50μg/kg),超标率为42.9%;其他6种抗生素检出均较低,且均未检出诺氟沙星。鲈鱼和鮰鱼肌肉组织中FQs残留的检出率最高,其中,氧氟沙星和恩诺沙星均检出;黄蜂鱼和黄搡鱼体内检出的残留量均较低,且抗生素残留种类也较少。结论贵阳市场的淡水鱼存在一定程度的FQs污染,应进一步关注氟喹诺酮类抗生素残留量的超标情况。     

烤鳗中氟喹诺酮类多残留检测方法的研究

目的:建立烤鳗中氧氟沙星、环丙沙星、恩诺沙星、双氟沙星、司帕沙星5种氟喹诺酮残留的高效液相色谱-质谱联用检测方法.方法:样品经磷酸盐缓冲液提取后,用固相萃取小柱进行净化、浓缩,Cloversil-C18柱(150 mm×4.6 mm i.d,5 μm)分离;流速:0.5 ml/min;流动相:甲醇/乙腈/0.2%甲酸(15/15/70,v/v/v);采用选择离子监测(SIM)技术在电喷雾电离(ESI)正离子模式下检测,质谱定量离子氧氟沙星、环丙沙星、恩诺沙星、双氟沙星、司帕沙星分别为m/z::361.8[M H] 、331.8[M H] 、359.8[M H] 、399.8[M H] 和392.8[M H] .结果:5种被测化合物在0.50～100.0μg/kg范围内均呈良好的线性关系,回收率大于98.6%,RSD均小于7.0%.检出限为0.5 μg/kg.结论:本法简便、灵敏、重现性好、特异性强,是烤鳗中5种氟喹喏酮残留检测的有效方法.     

高效液相色谱荧光检测法测定牛奶中3种氟喹诺酮残留研究

目的建立牛奶中环丙沙星、双氟沙星和沙拉沙星3种氟喹诺酮残留的高效液相色谱荧光检测方法.方法样品经磷酸盐缓冲液稀释、提取后,用Waters Oasis(R)HLB固相萃取小柱进行浓缩、净化,Extend-C18柱(250 mm×4.6 mm×5μm)分离;流速0.8 ml/min;柱温35℃;流动相甲醇 乙腈 0.2%甲酸(15 15 70);荧光检测波长λex为275 nm;λem为445 nm.结果在0.1～5.0 mg/L范围内三种被测化合物均呈良好线性关系,方法回收率在90.0%～102.0%之间,相对标准偏差为4.1%～10.0%,最低检出限环丙沙星、双氟沙星和沙拉沙星分别为0.02、0.015和0.02 mg/L.结论所建方法简便、灵敏、重现性好、干扰少、特异性好,能满足牛奶中三种目标化合物的检测要求.     

LC/MS/MS法同时测定食品中喹诺酮类药物

LC／MS／MS法同时测定食品中依诺沙星(Eno),氧氟沙星(Oflo),环丙沙星(Cip),达诺沙星(Dano),洛美沙星(Lome),恩氟沙星(Enro)和沙拉沙星(Sara)7种喹诺酮类药物。     

群"星"璀璨,当心伤人!

依诺沙星、诺氟沙星、环丙沙星、氧氟沙星……这些喹诺酮类抗生素以其广谱、高效、低毒,口服吸收完全,不能口服者可静脉滴注等优点,在临床上占有着越来越重要的地位。如依诺沙星(氟啶酸)、诺氟沙星(氟哌酸)、环丙沙星(环丙氟哌酸)、氟罗沙星、洛美沙星、芦氟沙星、氧氟沙星(氟嗪酸)、左氟沙星等是抗菌的主力,司帕沙星、妥苏沙星、加替沙星、格帕沙星、     

高效液相色谱-串联质谱法同时测定鱼、虾中18种抗生素药物残留

目的建立高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定鱼、虾中氯霉素、金霉素、土霉素、四环素、环丙沙星、诺氟沙星、培氟沙星、依诺沙星、氟甲喹、恶喹酸、恩诺沙星、二氟沙星、斯帕沙星、氧氟沙星、丹诺沙星、奥比沙星、沙拉沙星、氟罗沙星等18种抗生素的检测方法。方法样品经EDTA-Mcllvaine缓冲溶液超声提取,高速离心,滤液经正己烷脱脂,HLB固相萃取柱净化,氮气吹至近干。Poroshell 120 C18柱色谱分离,流动相为0. 1%甲酸水溶液-乙腈,梯度洗脱,流速为0. 4 ml/min,进样量为5μl,柱温为40℃。电喷雾正负离子ESI(+/-)扫描;动态多反应(DMRM)质谱监测;基质匹配内标法定量。结果 18种抗生素线性良好,相关系数 0. 998;方法检出限为0. 1μg/L～2. 5μg/L,3个加标水平的回收率在70. 52%～127. 45%,相对标准偏差(RSD)在1. 48%～15. 93%。结论该方法简便快速、灵敏可靠,适用于水产品中18种抗生素药物残留的同时测定。     

Assessment of the genotoxicity of quinolone and fluoroquinolones contaminated soil with the Vicia faba micronucleus test

The genotoxicity of quinolone and fluroquinolones was assessed using the micronucleus (MN) test on Vicia faba roots by direct contact exposure to a solid matrix. Plants were exposed to quinolones (nalidixic acid) and fluoroquinolones (ciprofloxacin and enrofloxacin) alone or mixed with artificially contaminated soils. Four different concentrations of each of these antibiotics were tested (0.01, 0.1, 1 and 10 mg/Kg) for nalidixic acid and (0.005, 0.05, 0.5 and 5 mg/Kg) for ciprofloxacin and enrofloxacin. These antibiotics were also used in mixture. Exposure of Vicia faba plants to each antibiotic at the highest two concentrations showed significant MN induction. The lowest two concentrations had no significant genotoxic effect. The mixture of the three compounds induced a significant MN induction whatever the mixture tested, from 0.02 to 20 mg/Kg. The results indicated that a similar genotoxic effect was obtained with the mixture at 0.2 mg/Kg in comparison with each molecule alone at 5-10 mg/Kg. Data revealed a clear synergism of these molecules on Vicia faba genotoxicity.     

苏州市水产品中喹诺酮类抗生素残留水平及安全性评价

目的:了解苏州地区水产品中喹诺酮类抗生素残留水平,探讨其污染来源,评价其膳食安全性。方法:2011年6月-2012年4月随机采集苏州地区各类水产品,采用超高效液相色谱/质谱进行喹诺酮类抗生素检测。结果:95件水产品中,29件样品检出喹诺酮抗生素残留,检出率30.5%,检出的种类主要为恩诺沙星,与其他几种相比,有显著性统计学差异(P<0.01)。水产品中检出的恩诺沙星残留最高90.6μg/kg,均小于最高残留限量100μg/kg,未超过每日安全摄入量120μg/d。结论:苏州市市售水产品部分存在抗生素残留,需要加强水产品养殖、销售的管理。     

Antibiotic susceptibility of campylobacter isolates from sewage and poultry abattoir drain water.

In this study, the in vitro susceptibility of 209 campylobacter strains to the quinolones nalidixic acid, flumequine, ciprofloxacin, enrofloxacin, and to ampicillin, tetracycline and erythromycin was tested by the disk diffusion method. The strains were isolated from poultry abattoir effluent (DWA) and two sewage purification plants (SPA and SPB). Sewage purification plant SPA received mixed sewage, including that from a poultry abattoir, whereas SPB did not receive sewage from any meat-processing industry. The quinolone resistance of the DWA isolates ranged from 28% for enrofloxacin to 50% for nalidixic acid. The strains isolated from the sewage purification plants were more susceptible to the quinolones with a range of 11-18% quinolone resistance for SPB isolates to 17-33% quinolone resistance for SPA isolates. The susceptibility criteria as recommended by National Committee Clinical Laboratory Standards (USA) cannot readily be employed for campylobacter isolates. This investigation shows that the resistance of campylobacter bacteria is highest in the plant receiving sewage from a poultry slaughterhouse. Monitoring of antibiotic resistance of aquatic Campylobacter spp. is important, as surface waters are recognized as possible sources of infection.     

Activity of nalidixic acid and fluoroquinolones on Escherichia coli strains isolated from non-complicated urinary infections (reseau TSN - France, 1999 - 2001)

OBJECTIVE: The authors studied the susceptibility of 1,647 non-repeat isolates of Escherichia coli to quinolones and fluoroquinolones. METHOD: The strains were isolated from non-complicated urinary infections in women 18-64 years of age. Data was provided by the TSN Database France, a real time electronic database which collects antibiotic susceptibility results and patient demographic data. The data was collected from 1999 to 2001 in 63 French hospital laboratories, each using their own routine test methods. Quantitative data was interpreted (S, I, R) according to CA-SFM breakpoint guidelines. RESULTS: Ninety-eight and 94,6 % of the strains were susceptible to ciprofloxacin and nalidixic acid respectively. Cross resistance was assessed as well as intrinsic difference in activity within the fluoroquinolone class. Current fluoroquinolones are still highly efficient, and ciprofloxacin is the most active. CONCLUSION: Since 1996, little change in resistance to fluoroquinolones has been observed. These results confirm the choice of fluoroquinolones as first intention therapy as recommended by consensus conferences.     

Toxicity of the fluoroquinolone antibiotics enrofloxacin and ciprofloxacin to photoautotrophic aquatic organisms

The present study investigated the growth inhibition effect of the fluoroquinolone antibiotics enrofloxacin and ciprofloxacin on four photoautotrophic aquatic species: the freshwater microalga Desmodesmus subspicatus, the cyanobacterium Anabaena flos-aquae, the monocotyledonous macrophyte Lemna minor, and the dicotyledonous macrophyte Myriophyllum spicatum. Both antibiotics, which act by inhibiting the bacterial DNA gyrase, demonstrated high toxicity to A. flos-aquae and L. minor and moderate to slight toxicity to D. subspicatus and M. spicatum. The cyanobacterium was the most sensitive species with median effective concentration (EC50) values of 173 and 10.2 μg/L for enrofloxacin and ciprofloxacin, respectively. Lemna minor proved to be similarly sensitive, with EC50 values of 107 and 62.5 μg/L for enrofloxacin and ciprofloxacin, respectively. While enrofloxacin was more toxic to green algae, ciprofloxacin was more toxic to cyanobacteria. Calculated EC50s for D. subspicatus were 5,568 μg/L and >8,042 μg/L for enrofloxacin and ciprofloxacin, respectively. These data, as well as effect data from the literature, were compared with predicted and reported environmental concentrations. For two of the four species, a risk was identified at ciprofloxacin concentrations found in surface waters, sewage treatment plant influents and effluents, as well as in hospital effluents. For ciprofloxacin the results of the present study indicate a risk even at the predicted environmental concentration. In contrast, for enrofloxacin no risk was identified at predicted and measured concentrations.     

Salmonella of Indian origin: pseudo-susceptibility to fluoroquinolones

We report the case of a patient presenting with typhoid fever after returning from a stay in India. This infection was not cured with a course of ciprofloxacin, due to a reduced susceptibility of the bacteria to the drug. This decreased susceptibility to fluoroquinolones was not detected by the antibiogram, but the MIC for nalidixic acid was greater than 32 mg/l. This case suggests using a third generation cephalosporin instead of a quinolone, for people coming from a high-risk zone. It also suggests that the MIC for nalidixic acid and for norfloxacin can be used as the first clue for a reduced susceptibility to fluoroquinolones.     

Campylobacter antimicrobial drug resistance among humans, broiler chickens, and pigs, France

We describe isolates from human Campylobacter infection in the French population and the isolates' antimicrobial drug resistance patterns since 1986 and compare the trends with those of isolates from broiler chickens and pigs from 1999 through 2004. Among 5,685 human Campylobacter isolates, 76.2% were C. jejuni, 17.2% C. coli, and 5.0% C. fetus. Resistance to nalidixic acid increased from 8.2% in 1990 to 26.3% in 2004 (p < 10(-3)), and resistance to ampicillin was high over time. Nalidixic acid resistance was greater for C. coli (21.3%) than for C. jejuni (14.9%, p < 10(-3)). C. jejuni resistance to ciprofloxacin in broilers decreased from 31.7% in 2002 to 9.0% in 2004 (p = 0.02). The patterns of resistance to quinolones and fluoroquinolones were similar between 1999 and 2004 in human and broiler isolates for C. jejuni. These results suggest a potential benefit of a regulation policy limiting use of antimicrobial drugs in food animals.     

铜绿假单胞菌的耐药性及其耐氟喹诺酮机制的研究

目的：了解铜绿假单胞菌的耐药性及其耐氟喹诺酮的机制。方法：用纸片扩散法测定626株铜绿假单胞菌对11种抗生素的耐药性。琼脂稀释法测定40株耐环丙沙星的铜绿假单胞菌对环丙沙星和左氧氟沙星的最低抑菌浓度（MIC），聚合酶链反应－限制性长度多态性分析检测耐环沙星的铜绿假单胞菌的gyrA基因和parC基因突变，结果：626株铜绿假单胞菌中耐环丙沙星的180株（28．8％），耐左氧氟沙星的219株（35．0％），40株耐环丙沙星的铜绿假单胞菌有30株（75％）发生gyrA基因83位点突变，26株（65％）发生parC基因87位点突变，两种突变同时发生的25株（62．5％），结论：铜绿假单胞菌的耐药性日趋严重，临床分离的铜绿假单胞菌耐氟喹诺酮的机制大多为药物作用靶位gyrA和parC的基因突变。     

8种氟喹诺酮类药物的体外敏感性

目的比较大肠埃希菌、铜绿假单胞菌和金黄色葡萄球菌对8种氟喹诺酮类药物的体外抗菌活性. 方法按NCCLS推荐的标准纸片扩散法(K-B法)对69株大肠埃希菌、84株铜绿假单胞菌和82株金黄色葡萄球菌进行8种三代氟喹诺酮药物的体外敏感性实验. 结果对于大肠埃希菌和铜绿假单胞菌,环丙沙星的体外抗菌活性最强;对于金黄色葡萄球菌,虽环丙沙星的敏感率低于氧氟沙星、左氧氟沙星及司帕沙星等,但并未提示差异有显著性(P>0.05). 结论 8种帕喹诺酮类药物显示明显的交叉耐药性,环丙沙星的体外敏感性结果对于临床应用三代氟喹诺酮类药物具有一定的指导意义.