高效液相色谱法测定鸡蛋中4种氟喹诺酮类 药物的含量

目的采用高效液相色谱法同时测定鸡蛋中环丙沙星、恩诺沙星、沙拉沙星和达氟沙星4种药物的残留量。方法样品经磷酸盐提取液提取后,经反相C_(18)柱分离,采用0.05 mol/L磷酸-三乙胺溶液:乙腈(85:15,V:V)流动相进行等度洗脱,流速为0.8 m L/min,采用荧光检测器在激发波长280 nm、发射波长450 nm的条件下进行测定,以外标法定量。结果环丙沙星、恩诺沙星和沙拉沙星在0.005~0.5μg/m L、达氟沙星在0.001~0.1μg/m L浓度范围内具有良好的线性关系,相关系数为0.9998,加标回收率为96.5%~103%,相对标准偏差为0.83%~1.28%。结论该方法精确可靠,重复性、稳定性和分离效果好,可适用于测定鸡蛋中氟喹诺酮类药物的残留量。     

鸡肉源沙门氏菌对(氟)喹诺酮类抗生素的耐药性及相关基因

目的:研究分离于广东、广西、福建和上海4省(市)零售鸡肉源沙门氏菌对萘啶酮酸和部分氟喹诺酮类抗生素的药敏性及相关耐药基因,以更好地了解耐药性的产生和传播途径,确保食品安全。方法:使用临床和实验室标准协会推荐的琼脂稀释法测定沙门氏菌的药敏性,用PCR、基因序列测定和基因库在线比对方法确定耐药沙门氏菌中与(氟)喹诺酮类抗生素耐药相关的gyr A亚基中喹诺酮类抗性决定区,par C亚基中的氨基酸突变状况以及qnr质粒携带的与喹诺酮类抗生素耐药相关基因。结果:358株沙门氏菌中,59.78%的菌株对萘啶酮酸产生抗性,对环丙沙星、左氧氟沙星和加替沙星耐药的菌株比例分别为22.91%,17.88%和16.20%。214株萘啶酮酸抗性菌中,qnr A,qnr B,qnr S和aac(6′)-Ib-cr基因的检出率分别为11.68%,18.22%,3.27%和24.77%。82株环丙沙星抗性菌中,从gyr A和par C基因中共检出135个氨基酸突变点,其中从gyr A基因中检出65个突变点,从par C基因中检出70个突变点。gyr A基因中以Asp87Asn突变最为常见(47.69%;31/82),其次分别为Asp87Gly(38.46%,25/82),Asp87Tyr(4.62%,3/82),Ser83Phe和Asp87Asn双突变(3.80%,1/82),Asp87Asn和Ile89Val双突变(3.80%,1/82),Asp87Asn和Val90Gly双突变(3.80%,1/82)。par C基因中Ser80Arg突变最为常见(90.00%,63/82),其次分别为Met118Ile-Arg119Leu-Thr121Ser三突变(4.29%,1/82),Ser80Arg和Tyr120Phe双突变(2.86%,1/82),Ser80Ile(1.43%,1/82)和Ala81Val(1.43%;1/82)。结论:4省市中鸡肉源沙门氏菌对萘啶酮酸和部分氟喹诺酮类抗生素耐药严重,其中解旋酶和拓扑异构酶基因突变以及沙门氏菌携带的qnr A,qnr B,qnr S和aac(6′)-Ib-cr基因是沙门氏菌耐药的重要原因。     

微波辅助萃取-液相色谱-串联质谱法检测猪肉中四种氟喹诺酮类药物残留

建立了微波辅助萃取-液相色谱-串联质谱法检测猪肉中氧氟沙星、诺氟沙星、环丙沙星、恩诺沙星四种氟喹诺酮类药物残留的方法。样品经微波辅助萃取,正己烷脱脂,无水硫酸钠除水,浓缩提取物至干,用1.0mL0.1%甲酸溶液溶解后上机检测。采用Hypersil GOLD-1.9μm,50 mm×2.1 mm(i.d)色谱柱分离,在多反应监测模式下检测,外标法定量。结果表明:四种氟喹诺酮类药物在浓度10～200μg/L范围内线性关系良好,相关系数r=0.9956～0.9991,定量限为10μg/kg。在不同添加水平下,其平均回收率为88%～101%,变异系数为1.4%～4.1%。     

Functional Gelatin-Carbon Nanotubes Nanohybrids With Enhanced Antibacterial Activity

Hybrid materials with enhanced antibacterial activity were prepared by incorporation of carbon nanotubes within gelatin-fluoroquinolones bioconjugates. Gelatin bioconjugates were characterized by UV-Vis, FT-IR, and calorimetric analyses, nanohybrids by morphological analyses. Biocompatibility was evaluated on human mesenchymal stem cells, and antibacterial performance against Klebsiella pneumoniae and Escherichia coli. Minimun inhibitory concentrations from 0.025 to 0.05 µg mL^?1 and from 0.025 to 0.10 µg mL^?1, and MBC from 0.025 to 0.10 µg mL^?1 and from 0.05 to 0.20 µg mL^?1 were detected for Escherichia coli and Klebsiella pneumoniae, respectively, showing that nanotubes increase antimicrobial activity comparing to both free and gelatin-conjugated drugs.     

A simple method for the determination of fluoroquinolone residues in tilapia (Oreochromis niloticus) and pacu (Piaractus mesopotamicus) employing LC-MS/MS QToF

The use of antimicrobials in livestock production is a powerful resource applied throughout the world to guarantee high yield and control bacterial diseases in aquaculture. However, residues of these substances in animal products represent a potential risk to consumer health when residue levels are above the established maximum residue limits (MRLs). Fluoroquinolones (FQs) are antimicrobials commonly used worldwide in aquaculture. The aim of this work was to develop and validate a simple analytical method for the simultaneous determination of norfloxacin, danofloxacin, enrofloxacin and ciprofloxacin levels in tilapia (Oreochromis niloticus) and pacu (Piaractus mesopotamicus) fillets using liquid chromatography–tandem mass spectrometry (LC-MS/MS) quadrupole time of flight (QToF). The FQs were extracted from the fillets with 1% acetic acid–methanol and 1% acetic acid–acetonitrile solutions using ultrasonic assistance. The clean-up was performed with hexane. Chromatographic separation was conducted in an XTerra RP18 column (2.1 × 150 mm, 5 µm) at 25 °C with a flow of 0.2 mL min^?1. The mobile phase consisted of 0.1% aqueous formic acid and acetonitrile, with gradient elution. The validation parameters for all FQs were linearity (>0.99), intra-day precision (CV of 1%–9%), inter-day precision (CV of 3%–17%), decision limit (63–126 ng g^?1), detection capability (76 –152 ng g^?1) and accuracy (90%–111%). The limit of quantification was lower than the MRL for each FQ, indicating that the method is suitable for the determination of the FQ levels in the fish fillets. The mass analyser of the QToF type was able to confirm the identities of the FQs with an error of the accuracy of the mass (reasons m/z) of less than 10 ppm.     

Gemifloxacin mesylate (GFM): dissolution test based on in vivo data

Gemifloxacin mesylate (GFM) is a synthetic, broad-spectrum, fluoroquinolone antibacterial agent. It is different from other class members because it achieves adequate plasma concentrations to inhibit both topoisomerase IV and gyrase. The aim of this study was to develop and validate a dissolution test for GFM in coated tablets, using a simulated absorption profile based on in vivo data obtained from the literature. The fraction and percentage of the dose absorbed were calculated using model-dependent Loo-Riegelman approach for two compartments. The best in vitro dissolution profile was obtained using 900?mL of pH 6.0 phosphate buffer as a dissolution medium at 37?°C?±?0.5?°C and paddles at 50?rpm. The in vitro dissolution samples were analyzed using a liquid chromatography method, and the validation was performed according to USP 34 (2011). The method showed specificity, precision, accuracy, robustness and linearity. Under these conditions, a level-A in vitro–in vivo correlation was suggested (r?=?0.9926). The prediction errors were calculated to determine the validity and accuracy of the suggested correlation. The dissolution test can be used to evaluate the dissolution profile of GFM-coated tablets and minimize the number of bioavailability studies as part of new formulation development.     

Determination of aminoglycosides and quinolones in food using tandem mass spectrometry: a review

The widespread use of antibiotics in dairy cattle management may result in the presence of antibiotic residues in food. While rapid screening tests are commonly used to detect the presence of antibiotics in food, more accurate chromatographic-mass spectrometric methods combined with tandem mass spectrometry (MS/MS) are required to determine the identity and quantity of the antibiotic present. These methods (HPLC/MS/MS) may have the greatest potential for accomplishing direct multi-residue identifications in complex biological matrices, such as food. This study reviews recent applications of tandem mass spectrometry in the determination of antibiotic residues, such as aminoglycosides and quinolones in food.     

Mechanism of binding of fluoroquinolones to the quinolone resistance-determining region of DNA gyrase: towards an understanding of the molecular basis of quinolone resistance

We have studied the bacterial resistance to fluoroquinolones that arises as a result of mutations in the DNA gyrase target protein. Although it is known that DNA gyrase is a target of quinolone antibacterial agents, the molecular details of the quinolone-gyrase interaction remain unclear. The mode of binding of ciprofloxacin, levofloxacin, and moxifloxacin to DNA gyrase was analyzed by means of docking calculations over the surface of the QRDR of GyrA. The analysis of these binding models allows study of the resistance mechanism associated with gyrA mutations more commonly found in E. coli fluoroquinolone-resistant strains at the atomic level. Asp87 was found to be critical in the binding of these fluoroquinolones because it interacts with the positively charged nitrogens in these bactericidal drugs. The role of the other most common mutations at amino acid codon Ser83 can be explained through the contacts that the side chain of this residue establishes with fluoroquinolone molecules. Finally, our results strongly suggest that, although Arg121 has never been found to be associated with fluoroquinolone resistance, this residue makes a pivotal contribution to the binding of the antibiotic to GyrA and to defining its position in the QRDR of the enzyme.     

Terbium-sensitised luminescence screening method for fluoroquinolones in beef serum

Enrofloxacin and danofloxacin are the only fluoroquinolone antibiotics approved for use in cattle in the United States. Microbial screening methods commonly used for monitoring veterinary drug residues are not sensitive or selective for fluoroquinolones. In this work, a luminescence-based screening assay was developed to detect fluoroquinolones in beef serum. This approach takes advantage of the DNA-enhanced luminescence signal of a fluoroquinolone–Tb⁺³ complex. In this method, serum samples were extracted with acidified acetonitrile in the presence of magnesium sulfate. After centrifugation, evaporation of the supernatant was followed by dissolution of the residue in buffer and filtration. Addition of Tb⁺³ and DNA then allowed a reading of the luminescence signal. The technique was illustrated using enrofloxacin, and provided good recoveries (73–88%) at 25, 50 and 100 ng ml^?1, with reasonable RSDs averaging at 11%. The LOD was 2.5 ng ml^?1 based on the variability of response of control serum samples from 18 different steers. The method provided no false-positive or false-negative results while screening blind samples for enrofloxacin and was demonstrated to be quantitative over a range of 0–100 ng ml^?1.     

水兼容性分子烙印固相萃取-高效液相色谱法测定鸡肉中氟喹诺酮类药物残留

目的：建立分子烙印固相萃取-高效液相色谱测定鸡肉中氟喹诺酮类药物残留的新方法。方法：以甲基丙烯酸为功能单体,恩诺沙星为模板分子,在强极性溶剂甲醇-水体系中制备分子烙印聚合物,考察和评价分子烙印聚合物的特性。鸡肉样品经乙腈提取浓缩后过分子烙印固相萃取柱,乙腈-三氟乙酸(99:1,V/V)洗脱液由高效液相色谱分离和荧光法检测。结果：高亲合力的结合位点的解离常数为Kd1=7.19×10-5 mol/L,最大表观结合位点数Qmax,1=110.19μmol/g;低亲和力结合位点的解离常数为Kd2=2.44×10-3mol/L,最大表观结合位点数Qmax,2 = 965.51μmol/g。氧氟沙星、诺氟沙星、环丙沙星和恩诺沙星等氟喹诺酮类药物的校准曲线在1.0～500 μg/kg范围内呈良好的线性关系(r≥0.9991),检出限(S/N=3)为0.06～0.09μg/kg,平均回收率为75.4%～86.4%(n=3),相对标准偏差小于6%。结论：以水兼容性分子烙印固相萃取-高效液相色谱法可实现氟喹诺酮药物的有效分离和灵敏测定。