An analytical method was developed for the simultaneous determination of penicillins (benzylpenicillin, oxacillin, cloxacillin, dicloxacillin and nafcillin) remaining in edible tissues (muscle and kidney) of animals (bovine and swine) by pre-columnfluorescence derivatization. For labeling a carboxyl group in penicillins, seven kinds of fluorescence derivatization reagents were tested: 3-bromomethyl-6,7-dimethoxy-1-methyl-1,2-dihydroquinoxaline-2-one (Br-DMEQ), 4-bromomethyl-7-methoxycoumarin (Br-Mmc), 5,6-dimethoxy-2-(4-hydrazinocarbonylphenyl) benzothiazole (BHBT), 1-bromoacetylpyrene (BAP), 6,7-dimethoxy-1-methyl-2(H)-quinoallnone-3-propinohydrazide (DMEQ-hydrazide), 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate (NE-OTf) and 9-anthryldiazomethane (ADAM). BAP showed the best score on reactivity and was selected and used for the following examinations. Optimum conditions for HPLC analysis were determined with reference to the stability of each reagent, reactive temperature and time. Edible animal tissues were extracted with water and deproteinized with sulfuric acid and sodium tangstate, followed by concentration with C18 (solid-phase extraction) cartridge column. The acetonitrile eluate from the column was derivatized with BAP and 18-crown-6 reagents at 40 deg C for 30 min. The derivatized sample was analyzed by HPLC with reversed-phase mode. The mean recovery range was 73-96 percent for benzylpenicillin, 73-90 percent for oxacillin, 64-83 percent for cloxacillin, 62-71 percent for nafcillin and 61-67 percent for dicloxacillin in tissues. The detection limits for benzylpenicillin and other penicillins were about 2 and 5 ng/g in tissues, respectively. In the suspected case of penicillin residues, benzylpenicillin was determined (0.045-29.0 mu m g/g) in bovine muscle and kidney by this method. The simple and sensitive method was expected for routine analysis of penicillins in edible animal tissues.
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机译：开发了一种分析方法，用于通过预柱荧光衍生化在动物（牛和猪）的食用组织（肌肉和肾脏）中留下青霉素（苄基百霉素，牛奶菌素，克罗克林，双氧化嘧啶和Nafcillin）。为了在青霉素中标记羧基，测试了七种荧光衍生化试剂：3-溴甲基-6,7-二甲氧基-1-甲基-1,2-二氢吲哚-1-甲基-1,2-二羟基毒 - 2-一（BR-DMEQ），4-溴甲基 - 7-甲氧基苏马林（BR-MMC），5,6-二甲氧基-2-（4-肼羰基苯基）苯并噻唑（BHBT），1-溴乙酰芘（BAP），6,7-二甲氧基-1-甲基-2（H）-Quinoallnone -3-丙基酰肼（DMEQ-酰肼），2-（2,3-萘氨基）三氟甲磺酸乙酯（NE-OTF）和9- Anthryldiazomethane（ADAM）。 BAP显示了反应性的最佳分数，并选择并用于以下考试。参考每种试剂，反应温度和时间的稳定性测定HPLC分析的最佳条件。用水萃取可食用的动物组织并用硫酸和橘子剥夺，然后用C18（固相萃取）盒柱浓度。从塔中的乙腈洗脱液用烤盘和18℃-6试剂以40℃衍生30分钟。通过HPLC分析衍生化的样品，具有反相模式。苄基培素的平均恢复范围为73-96％，氧化苯胺蛋白为73-90％，克罗克里林蛋白的64-83％，Nafcillin的62-71％，组织中双氯甲嘧啶的61-67％。苄基百年林和其他青霉素的检测限分别在组织中为约2和5 ng / g。在疑似案例的青霉素残留情况下，通过该方法测定伯齐尼霉素（0.045-29.0μmg/ g）（0.045-29.0 mu m g / g）。预期简单敏感的方法对食用动物组织中青霉素的常规分析。
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