高碘酸钾-鲁米诺后化学发光反应的研究——流动注射-后化学发光法测定异烟肼

实验发现了异烟肼在高碘酸钾-鲁米诺化学发光反应体系中的后化学发光反应。在对这一后化学发光反应的动力学性质、化学发光光谱、荧光光谱、紫外可见吸收光谱以及一些相关问题研究的基础上,讨论了其可能的反应机理;建立了利用此后化学发光反应测定异烟肼的流动注射化学发光新方法。方法的线性范围为1.0×10-9~4.0×10-7g/mL,检出限为6×10-10g/mL,对5.0×10-8g/mL的异烟肼进行11次平行测定的相对标准偏差为1.7%。此法已用于异烟肼片剂中异烟肼含量的测定,结果与药典法测定值一致。     

荧光猝灭法测定药物中异烟肼含量

在pH 6.5的Walpole缓冲溶液中,当以300 nm的光激发时,1,2-萘醌-4-磺酸钠在468 nm处产生1个较强的荧光峰。加入异烟肼后,由于其与1,2-萘醌-4-磺酸钠发生反应,从而使体系颜色加深且荧光信号减弱。在最佳实验条件下,在0.10~8.0μg/mL范围内,体系的荧光信号△F与异烟肼浓度之间有较好的线性关系,检出限为40.2 ng/mL。方法可直接用于测定药物中异烟肼含量,回收率为96.5%~106.3%。     

罗丹明类染料荧光猝灭法测定异烟肼

在HAc-NaAc缓冲溶液中,罗丹明类染料罗丹明6G(Rh6G)、罗丹明B(RhB)和丁基罗丹明(b-RhB)分别在553,582和589 nm处产生一个较强的荧光峰。加入异烟肼(INH)后,由于异烟肼分别与Rh6G,RhB和b-RhB发生作用,从而使得3种体系的荧光均发生猝灭。在最佳实验条件下,异烟肼浓度分别在0. 10～20μg/mL,0. 10～25μg/mL和0. 50～22μg/mL范围内与各体系的荧光强度改变值(!F)之间呈良好的线性关系,且3种体系的检出限分别为9. 04,21. 7和20. 4 ng/mL。据此,建立了一种荧光猝灭法测定药物中异烟肼含量的分析方法。     

流动注射后化学发光法测定异烟肼

本文发现了异烟肼在铁氰化钾-钙黄绿素化学发光反应体系中的后化学发光反应。优化了反应条件,建立了一种利用后化学发光反应测定异烟肼的流动注射化学发光分析法。方法的检出限为6×10-8g/mL, 相对标准偏差为1.8% (2.0×10-6 g/mL 异烟肼,n=11）,线性范围为2.0×10-7~1.0×10-5 g/mL。此法已用于异烟肼片剂中异烟肼含量的测定,结果与药典方法测定值一致。     

同步荧光光谱法直接测定尿液中卡那霉素

基于卡那霉素对CdTe量子点荧光的增强效应,建立一种直接测定尿液中卡那霉素含量的同步荧光光谱法。研究发现采用同步荧光光谱法,能够有效避免尿液中复杂基质的荧光干扰。在优化实验条件下,卡那霉素浓度在0.2~20.0μg/mL范围内与体系相对同步荧光强度呈良好的线性关系,相关系数为0.9996,检出限为0.005μg/mL。该方法用于尿液样品中卡那霉素含量的直接测定,回收率为96.3%~104.4%。     

毛细管电泳-化学发光法测定人血清中的异烟肼

基于碱性介质中异烟肼对laminol-K3Fe(CN)6化学发光体系的增敏作用,设计了一个经毛细管电泳(CE)分离,在线化学发光检测异烟肼的新方法.研究并优化了毛细管电泳分离及化学发光检测的条件.在优化的实验条件下,该方法测定异烟肼的线性范围为4.0×10-7～1.0×10-5g/mL(R2=0.9984),检出限(3σ)为1×10-8g/mL,对6.0×10-6g/mL的异烟肼进行6次平行测定,其相对标准偏差为4.0%.方法已用于血清中异烟肼的测定.     

离子液体/表面活性剂协同增敏荧光法测定米力农

基于米力农自身荧光在十六烷基三甲基溴化铵(CTAB)和室温离子液体(RTILs)溴化十二烷基三甲基咪唑混合体系中得到显著增强,提出了测定药物及生物体液中米力农含量的荧光新方法。在优化条件下,线性方程为F=47.32+1378.65ρ(ρ:μg/mL,r=0.998),线性范围为0.02~1.5μg/mL,检出限为6.4 ng/mL。实验中对荧光增敏机理进行了探讨。将方法用于米力农注射液和人血清、尿液中米力农含量的测定,回收率在97.5%~107.4%,米力农注射液测定结果与法定方法一致。     

金纳米粒子催化鲁米诺-异烟肼化学发光反应及其分析应用

研究发现, 在金纳米粒子存在下, 异烟肼与鲁米诺反应可产生强的化学发光. 采用响应曲面优化法中的Box-Behnken设计, 对试剂浓度进行了优化. 在优化的实验条件下, 相对化学发光强度与异烟肼的质量浓度在5.0× 10^－9～3.0×10^－7 g/mL范围内呈线性关系. 方法的检出限为4.1×10^－9 g/mL异烟肼, 相对标准偏差为1.3% (1.0×10^－7 g/mL异烟肼溶液, n＝11). 该方法已用于片剂、注射液和人血清中异烟肼含量的测定. 同时, 对化学发光反应的可能机理也进行了讨论.     

酶催化光度法测定异烟肼

基于异烟肼对牛血红蛋白模拟酶催化体系的抑制作用,建立了酶催化光度法测定异烟肼的新方法。研究了该抑制反应的最佳实验条件及动力学行为,测定的线性范围为3.65×10-8～3.65×10-6mol/L,检出限为5.4×10-9mol/L。对浓度为1.27×10-6mol/L的异烟肼进行11次平行测定,其相对标准偏差为3.9%。该方法可用于异烟肼注射液中异烟肼含量的测定。     

共振瑞利散射光谱法测定异烟肼

基于在PH=1.5的CL缓冲溶液中异烟肼的加入使曙红Y的共振瑞利散射信号增强,建立了一种异烟肼测定方法.异烟肼浓度在0.02～4.2μg/mL范围内与体系散射强度的增强呈线性关系,检出限为0.01851μg/mL.该方法用于异烟肼片剂的测定,结果满意.     

Flow-injection chemiluminescence sensor for determination of isoniazid in urine sample based on molecularly imprinted polymer

In this paper, molecularly imprinted polymer (MIP) of isoniazid is synthesized through thermal radical copolymerization of metharylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) in the presence of isoniazid template molecules. A novel flow injection chemiluminescence sensor for isoniazid determination is developed by packing the isoniazid-MIP into the flow cell as recognition elements. Isoniazid could be selectively adsorbed by the MIPs and the adsorbed isoniazid was sensed by its great enhancing effect on the weak CL reaction between luminol and periodate which were mixed in the flow cell. The enhanced CL intensity is linear in the range 2x10(-9) to 2x10(-7) g/mL and the detection limit is 7x10(-10) g/mL (3sigma) isoniazid with a relative standard deviation 2.8% (n=9) for 8x10(-8) g/mL. The sensor is reversible and reusable. It has a great improvement in sensitivity and selectivity for CL analysis. As a result, the sensor has been successfully applied to determination of isoniazid in human urine. At the same time, the binding characteristic of the polymer to isoniazid was evaluated by batch method and the dynamic method, respectively.     

Flow Injection Chemiluminescence Determination of Isoniazid Using On-Line Electrogenerated Manganese(III) as Oxidant

A novel flow injection chemiluminescence (CL) system for the determination of isoniazid has been proposed. It is based on the direct CL reaction of isoniazid and Mn(III) in sulfuric acid medium. The unstable Mn(III) was on-line electrogenerated by constant current electrolysis. The CL emission intensity was linear with isoniazid concentration in the range 0.1–10 μg/mL; the detection limit was 3.2 × 10⁻² μg/mL. The whole process could be completed in 1 min with a relative standard deviation of less than 5%. The proposed method is suitable for automatic and continuous analysis and has been applied successfully to the analysis of isoniazid in pharmaceutical preparation.     

磺胺嘧啶钠探针分析生物样品中蛋白质含量的研究

在pH 7.4的Tris-HCl缓冲溶液中,荧光猝灭光谱和三维荧光光谱显示磺胺嘧啶钠(SDS)可与人血清白蛋白(HSA)相互作用,使人血清白蛋白疏水微环境极性以及构象发生变化。考察了Δλ值、反应介质、离子强度等因素对该体系同步荧光光谱特征及强度的影响。在选定的最佳实验条件下,体系的同步荧光强度(ISF)与人血清白蛋白在1.38～276μg/mL的浓度范围内呈现良好的线性关系,线性相关系数为0.9992,从而建立了以磺胺嘧啶钠为分子探针,用固定波长同步荧光光谱分析测定蛋白质的新方法,检测限可达0.48μg/mL。用此方法对生物样品人血清、唾液和尿液中蛋白质含量进行了测定并进行加标回收实验,回收率在95.7%～103.7%之间。本文还用经典的考马斯亮蓝法对样品进行了测定,所得结果和本方法一致。同时还考察了一些常见的共存物质对蛋白质测定的影响。     

Simultaneous determination of first-line anti-tuberculosis drugs and their major metabolic ratios by liquid chromatography/tandem mass spectrometry

Monitoring of anti-tuberculosis drug concentrations and dose adjustment can be helpful in cases that show poor response to treatment. Here, we describe a method that can rapidly and simultaneously measure the blood concentrations of four anti-tuberculosis drugs (isoniazid, rifampicin, pyrazinamide, and ethambutol) and two major metabolic ratios (acetylisoniazid/isoniazid and 25-desacetylrifampicin/rifampicin) using high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS). A C18 reversed-phase column and gradients of methanol in 0.3% formic acid and water were used for HPLC separation. The drug concentrations were determined by multiple reaction monitoring in positive ion mode and the assay performance was evaluated. We determined peak concentration ranges for each drug and acetylisoniazid/isoniazid and 25-desacetylrifampicin/rifampicin ratios by analyzing 2-h post-dose samples in patients treated with standard dosing as a first-line treatment. The preparation of 20 samples including two steps of deproteinization with 50% and 100% methanol was performed within 20 min and chromatographic separation was achieved within 4 min/sample. Interassay calibration variability data obtained over concentrations of 0-8 microg/mL for isoniazid and ethambutol and 0-80 microg/mL for rifampicin and pyrazinamide showed a linear and reproducible curve. Within-run and between-run imprecision (CVs) were 1.9-5.5% and 3.5-10.5% and the lower limits of detection and quantification were 0.01-0.5 microg/mL and 0.05-1.0 microg/mL, respectively. The isoniazid concentration was found to be inversely correlated to the acetylisoniazid/isoniazid ratio (R=-0.739, P<0.001). The devised method allows for the simple, rapid, sensitive and reproducible quantification of isoniazid, rifampicin, pyrazinamide, ethambutol and their two metabolic ratios and should be helpful for therapeutic drug monitoring in tuberculosis patients.     

基于与铁氰化钾和Zn~(2+)作用共振光散射法测定异烟肼

在酸性介质中,异烟肼能将铁氰酸根离子还原成亚铁氰酸根离子,后者与硫酸锌反应生成K2Zn3[Fe(CN)6]2粒子引起体系的共振光散射信号显著增强.在345nm处增强的散射信号强度ΔIRLS与异烟肼的浓度在0.01~1.0μg/mL范围内呈线性关系,据此建立了一种检测异烟肼含量的共振光散射(RLS)分析方法.线性回归方程为ΔIRLS=136.1+4239c(c,μg/mL),相关系数(r)为0.9984,检测限(3σ)为3.8ng/mL.该方法已成功用于异烟肼片剂及血清样品的测定.此外,文中还结合吸收光谱,动态光散射,扫描电子显微镜等表征手段对反应机理和RLS信号强度增强的原因进行了探讨.     

红景天苷的荧光光谱分析

建立了测定红景天苷的荧光光谱方法.选择了最佳荧光激发波长和发射波长,考察了乙醇浓度、温度、放置时间、pH值和共存离子对测定结果的影响.红景天苷的荧光光谱具有良好的稳定性,在最佳实验条件下,其荧光强度与浓度在0.04 -18.03 μg/L范围内呈良好的线性关系,相关系数为0.9996,检出限为0.019 μg/mL.红...     

胶束增敏荧光法测定锆

在CTAB胶束介质中配合物组成可发生一系列变化,这是其对吸光度及荧光增敏的决定因素。在合适的CTAB浓度下,可形成Zr(Ⅳ):Morin=1:3的高次配合物,使体系的摩尔吸光系数增加3倍多;但随着CTAB浓度的增大,形成了Zr(Ⅳ):Morin:SO₄^2-=1:1:2的混配配合物,硫酸根起了辅助配体的作用,荧光强度增强数十倍,稳定性也有很大提高,对CTAB胶束改变配合物组成及增敏荧光的原因进行了研究。据此建立了胶束增敏荧光测定锆的新方法。该法的检测下限为0.2ng/mL,线性范围为0.2ng/mL～0.5μg/mL,选择性、稳定性均好。     

卤代荧光素荧光猝灭法测定肾上腺色腙

在pH=6.6 的缓冲介质中, 肾上腺色腙(CBZC)与二氯荧光素(DCF)、二溴荧光素(DBF) 和二碘荧光素(DIF)通过静电引力、芳基堆积作用和范德华力形成摩尔比为2: 1的复合物, 引起上述二卤代荧光素的荧光发生猝灭, 最大猝灭波长分别位于533, 536和560 nm. 其荧光猝灭值(ΔF)在一定范围内与肾上腺色腙浓度成正比, 荧光猝灭反应具有较高的灵敏度, 对CBZC的检出限分别为3.3 ng/mL(CBZC-DCF体系), 5.7 ng/mL(CBZC-DBF体系)和129.6 ng/mL(CBZC-DIF体系). 考察了共存物质的影响、荧光猝灭反应的适宜条件和影响因素, 结果表明, 该方法具有良好的选择性, 可用于CBZC的血药和尿药浓度的快速检测. 从温度的影响、荧光寿命以及Stern-Volmer图判断该反应为静态猝灭反应.     

A highly sensitive fluorimetric method for the determination of fluoride in biological material with Al3+-calcein complex

A highly sensitive fluorimetric method for the determination of fluoride was established. The method was based on quenching of the fluorescence of the Al3+-calcein complex in CCl3COOH-CH3COOK buffer solution by fluoride. The fluorescence emission was measured at lambdaex/lambdaem 480/503 nm, and the experimental variables and interference in this determination were studied. The linear calibration range was 0.8 ng/mL to 150 ng/mL and the detection limit was 0.2 ng/mL. The method was applied to determine fluoride in biological materials. The recovery was in the range of 96.3% to 104.7% and the relative standard deviation was 4.6%. The results obtained from the certified reference material by the present method agreed with the certified values.