荧光素二聚体荧光探针测定卡那霉素的研究

基于卡那霉素对荧光素二聚体具有良好的荧光增强作用,以现场形成的荧光素二聚体作为荧光探针,建立了卡那霉素定量分析的新方法。在碱性条件下（pH=9.0）,当溴化十六烷基三甲基铵（CTAB）浓度为0.4-1.0 mmol/L时,荧光素（FL）在CTAB作用下形成弱荧光现场二聚体,导致荧光猝灭。加入卡那霉素,体系荧光明显增强,且荧光强度与卡那霉素浓度呈良好的线性关系。在优化条件下,卡那霉素的测定线性范围为2.5×10^-3 - 0.1 g/L,检出限（3σ/K）为1.4×10^-4g/L。用于实际样品分析,测定回收率为101%-116%,RSD为1.9%-2.6%。     

Reversed-phase liquid chromatographic method for the analysis of aminoglycoside antibiotics using pre-column derivatization with phenylisocyanate

A pre-column derivatization liquid chromatographic method has been developed for the analysis of aminoglycoside antibiotics using phenylisocyanate as a derivatization reagent. Derivatives including kanamycin, neomycin and gentamicin were formed by reaction of the analytes with phenylisocyanate in the presence of triethylamine. Phenylisocyanato groups were attached to corresponding amino groups of aminoglycoside and their molecular mass was confirmed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The experimental conditions for derivatization and separation of aminoglycoside derivatives were optimized and validated. A simple liquid chromatographic method for the determination of aminoglycoside antibiotics was demonstrated.     

金纳米微粒作探针共振瑞利散射光谱法测定卡那霉素

在一种含柠檬酸盐的溶液中, 柠檬酸根阴离子自组装于带正电荷的金纳米微粒表面, 使金纳米微粒成为一种被柠檬酸根包裹的带负电荷的超分子化合物. 在pH 4.4～6.8的弱酸性介质中, 它可与质子化的卡那霉素(KANA)阳离子借静电引力、疏水作用力结合, 形成粒径更大的聚集体(平均粒径从12增至20 nm), 这种聚集体的形成在引起金纳米的等离子体吸收带明显红移(Δλ＝102 nm)的同时, 共振瑞利散射(RRS)显著增强并且倍频散射(FDS)和二级散射(SOS)等共振非线性散射也有较大的增强, 最大散射峰分别位于280 nm (RRS), 310 nm (FDS)和480 nm (SOS)处. 在适当条件下, 散射强度(ΔI)与卡那霉素的浓度成正比, 其中RRS法灵敏度最高, 因此金纳米微粒可作为测定卡那霉素的高灵敏RRS探针, 它对卡那霉素的检出限为10.52 ng•mL^－1, 方法有较好的选择性, 可用于血液中卡那霉素的测定, 文中还讨论了有关反应机理和RRS增强的原因.     

卡那霉素作为手性选择剂的毛细管电泳手性药物分离研究

建立了一种以天然易得的卡那霉素为手性添加剂，用毛细管区带电泳法快速分离市售对乙肝有良好治疗效果的药物联苯双脂衍生物的方法，拓宽了毛细管电泳中手性选择剂的范围，通过实验研究了卡那霉素、甲醇 含量PH值，磷酸盐缓冲体系和硼硝缓冲体系对手性分离的影响，以及三种有机溶剂（甲醇、乙晴、异丙醇）添加剂对手性分离的影响，结果表明，在含有3％卡那霉素，30mol/L，硼砂缓冲体系（PH＝8．0）添加30％异丙醇是最佳的分离条件。     

Microcalorimetric Evaluation of the Effect of Kanamycin： An Analysis Based on the Median-Effect Principle

A study of the effect of drug, kanamycin, on the growth metabolism of recombinant Escherichia coli B 1 was carded out by microcalorimeter monitoring of the metabolic activity of treated cells. Power-time curves of growing recombinant Escherichia coli cell suspensions, treated with different kanamycin doses, were recorded. The extent of the effect was evaluated by changes in the slopes of the microcalorimetric curves and the kinetics of the drug action was interpreted from the time at which these changes reached their maximum values and maintained their maximum values. Experimental dose-effect relationships conform to the median-effect principle of the mass-action law： fa/（1-fa）=（D/D50）^m. A plot of y=lg[（fa）^1-1]^-1 versus x=lg D gives the slope m, D50 and R∞. The experimental results revealed that high concentration of kanamycin had an inhibitory effect on the growth of recombinant Escherichia coli B 1 in the lg phase, and had a promoting effect in the stationary period. Moreover, it was demonstrated that microcalorimetry was a reliable method for the detection of modulatory effects in biology.     

HPLC‐ELSD determination of kanamycin B in the presence of kanamycin A in fermentation broth

A novel method for the direct determination of kanamycin B in the presence of kanamycin A in fermentation broth using high performance liquid chromatography with evaporative light scattering detector (HPLC‐ELSD) was developed. An Agilent Technologies C₁₈ column was utilized, evaporation temperature of 40°C and nitrogen pressure of 3.5 bar, the optimized mobile phase was water–acetonitrile (65:35, v/v), containing 11.6 mm heptafluorobutyric acid (isocratic elution with flow rate of 0.5 mL/min) with the gain 11. Kanamycin B was eluted at 5.6 min with an asymmetry factor of 1.827. The method showed good linearity over the concentration range of 0.05 to 0.80 mg/mL for the kanamycin B (r² = 0.9987). The intra‐day and inter‐day coefficients of variation obtained from kanamycin B were less than 4.3%. Mean recovery of kanamycin B from spiked fermentation broth was 95%. The developed method was applied to the determination of kanamycin B without any interference from other constituents in the fermentation broth. This method offers simple, rapid and quantitative detection of kanamycin B. Copyright © 2014 John Wiley & Sons, Ltd.     

核酸修饰的金纳米粒子用于分光光度法检测卡那霉素

建立了一种基于核酸修饰的金纳米粒子(Au NPs)检测卡那霉素的方法。该方法利用卡那霉素与适配体的特异性结合,游离适配体的部分互补序列,诱导核酸修饰的Au NPs聚集。通过对实验条件进行优化,结果表明在25℃条件下,适配体与其部分互补序列杂交摩尔比为1:1,与目标卡那霉素的作用时间1 h,加入核酸修饰的Au NPs反应2 h时,该方法的线性检测范围为6.3~43.8 nmol/L,检测限为5.3 nmol/L。将该方法应用于牛奶样品中卡那霉素的检测,回收率在95.1%~104.6%之间。     

Determinationation of kanamycin by square-wave cathodic adsorptive stripping voltammetry

A square-wave cathodic adsorptive stripping voltametric (SWCASV) method for the determination of kanamycin was developed on a thin-film mercury electrode (TFME). The optimal working conditions for the application of the method were found to be pH 8.0, provided by a Britton-Robinson (B.R.) buffer, and an adsorption potential of +0.30 V during 300 s. The equilibration time was applied during 10 s, and potential scans were performed at a scan rate of 40 mV/s, with a square-wave frequency of 100 Hz. The measuring-system response was linear over the kanamycin concentration range from 1.2 × 10⁻⁹ to 5.0 × 10⁻⁸ M and the detection limit achieved was 4.8 × 10⁻¹⁰ M. The relative error and relative standard deviation obtained were 1.20 and 4.67%, respectively. The voltammetric procedure was applied successfully to give a rapid and precise assay of kanamycin in kanamycin sulfate injection form. Published in Elektrokhimiya in Russian, 2008, Vol. 44, No. 12, pp. 1433–1437. The text was submitted by author in English     

鱼精蛋白-核酸适配体-金纳米技术快速检测牛奶中的卡那霉素

基于聚阳离子鱼精蛋白与带负电的核酸适配体以及金纳米粒子之间的静电作用,发展了一种生物纳米检测技术,用于卡那霉素的检测;优化了缓冲溶液中阳离子、鱼精蛋白以及核酸适配体浓度,结果表明在20 mmol/L Na~+、1 mmol/L Mg~(2+)、2 mg/L鱼精蛋白、100 nmol/L核酸适配体条件下,卡那霉素在5~5 000 nmol/L范围内与金纳米粒子的吸光度比值呈现良好的线性关系,相关系数(R2)为0.992 8,方法的检出限为0.53 nmol/L。在此实验条件下,检测了牛奶中卡那霉素的含量,回收率为96%~98%,相对标准偏差为1.5%~3.2%。该方法选择性高,灵敏度好,线性范围广,显示出其应用于食品中卡那霉素检测的优势。     

Label-Free and Simple G-quadruplex-based Turn-Off Fluorescence Assay for the Detection of Kanamycin

We have developed a label-free and turn-off fluorescence assay for the determination of kanamycin. The detection system consists of an aptamer for specifically recognizing kanamycin and two auxiliary probes functionalized with two GGG repeats at the 3′ or 5′ ends for signal reporting. Two probes both hybridize with the aptamer and then their G-rich sequences combine to form a G-quadruplex. When thioflavin T, a fluorophore, is bound to the G-quadruplex, the fluorescence intensity of the solution dramatically increases. Upon the addition of the kanamycin, the aptamer–kanamycin binding inhibits the hybridization of two probes and aptamer, and restrains the GGG repeats from getting closer to form the G-quadruplex structure, resulting a significant decrease in the fluorescence intensity. The proposed aptamer-based fluorescent sensing platform showed a linear relationship with the concentration of kanamycin from 0.6 to 20.0?nM. The detection limit was determined to be 0.33?nM. The sensing platform provides resistance to interferences from other antibiotics and can be used to efficiently recognize kanamycin in real samples.