Development of a modified beta-cyclodextrin based fluorosensor for dipyridamole

2,6-O-diethyl-β-cyclodextrins were prepared and immobilized onto silica gel to develop a fluorosensor for dipyridamole. The analytical performance characteristics of the proposed sensor for analysis of dipyridamole were as follows: the detection limit was 0.8 nmol/L with a relative standard deviation of 1.4% for 10 determinations of 50 nmol/L of dipyridamole. The modification of β-CD improved the sensitivity and selectivity for measuring dipyridamole. The recommended method has been successfully tested for the determination of dipyridamole in clinical samples (urine and pharmaceutical preparations). Received: 23 January 1997 / Revised: 14 April 1997 / Accepted: 21 April 1997     

双嘧达莫在玻碳电极上的阳极伏安法研究

用直流伏安法(DCV),微分脉冲伏安法(DPV)和循环伏安法(CV)在玻碳电极(GCE)上研究了双嘧达莫(DPM)在不同介质中的阳极伏安行为,发现在0.01mol/LCHl溶液中于0.62V(vs.Ag/AgCl)右左生产一个阳极氧化峰,其峰高与浓度的线性方程分别为r=0.0004 0.17C(0.05~1.0mg/L)和y=0.052 0.22C(0.5~10mg/L). 在此体中大多数金属离子和20多种有机生代物质不干扰测定,采用该方法可不经分离,直接测定药物制剂和尿样吕DPM的含量,同时对DPM的电极反反应机理进行了探讨.     

Gas chromatography/negative-ion chemical ionisation mass spectrometry for the quantitative analysis of morphine in human plasma using pentafluorobenzyl carbonate derivatives

A sensitive and specific method for the quantitative determination of morphine in human plasma is presented. Morphine was extracted from plasma by solid phase extraction on C18 and converted to its pentafluorobenzyl carbonate trimethylsilyl derivative. The derivatives were analysed without further purification. Using gas chromatography/negative ion chemical ionisation mass spectrometry, a useful diagnostic fragment ion at m/z 356 is obtained at high relative abundance. Deuterated morphine was used as internal standard. Calibration graphs were linear within the range 1.25 to 320 nmol/L. Intra-day precision was 3.82% (15 nmol/L), 2.85% (75 nmol/L) and 4.13% (225 nmol/L), inter-day variability was found to be 1.77% (15 nmol/L), 4.95% (75 nmol/L) and 9.88% (225 nmol/L). Inter-day accuracy showed deviations of 2.18% (15 nmol/L), -0.72% (75 nmol/L) and -0.13% (225 nmol/L). The method is rugged and robust and has been applied to the batch analysis of morphine during pharmacokinetic profiling of the drug.     

An inter-laboratory comparison for determination of cortisol in saliva

An inter-laboratory comparison study for cortisol in saliva is reported in the present study. Nine laboratories representing four different methods participated in the study. Each laboratory measured five blind samples prepared from natural saliva spiked with a pure certified reference material in the range 0–70 nmol/L. The average observed values were established as means of eight laboratories, because one laboratory was excluded as an outlier. The natural content of cortisol in the pooled saliva was 6.8 nmol/L. The recovery of the certified reference material was 83%–131% (164% for the outlying laboratory). For five laboratories the 95%-confidence intervals of the recoveries did not cover 100%. The standard deviations for a concentration of 10 nmol/L were estimated for each laboratory. They were 0.64–1.67 nmol/L (6.26 nmol/L for the outlying laboratory). The present study emphasizes the need for an external proficiency-testing scheme and a certified reference material for cortisol in saliva. Received: 8 May 2002 Accepted: 3 September 2002     

基于核壳金@铂纳米粒子的Ag~+比色检测方法

金核铂壳纳米粒子(Au@Pt NPs)具有出色的类过氧化物酶活性,而Ag~+对其催化活性表现出强烈的抑制;基于此,构建了高灵敏的Ag~+比色检测方法。在最佳反应条件下,Au@Pt NPs比色检测Ag~+的线性范围为0.1~10 nmol/L,检出限可达0.05 nmol/L。该方法对汞离子(Hg2~+)也表现出高灵敏的响应,比色检测Hg2~+的线性范围为10~200 nmol/L。将其应用于实际水样中Ag~+的检测,在添加浓度为10,50,100 nmol/L时,回收率为83.8%~97.7%,相对标准偏差为3.0%~9.6%,该方法具有操作简单、灵敏度高、成本低等优点。     

在线衍生-高效液相色谱法同时测定血浆中的色氨酸及其代谢物

建立了醋酸锌在线衍生高效液相色谱法同时测定血浆中色氨酸(Trp)、犬尿氨酸(Kyn)、5-羟吲哚乙酸(5-Hiaa)和犬尿喹啉酸(Kyna)的方法。以3-硝基酪氨酸为内标(IS),采用Hypersil C-18柱(250 mm×4.0 mm, 5 μ m),以250 mmol/L醋酸锌溶液(pH 5.5)-乙腈(95:5, v/v)为流动相,流速为0.8 mL/min,柱温30℃。荧光检测波长设定:5-Hiaa为278 nm(λ_ex)/343 nm(λ_em), Kyna为244 nm(λ_ex)/400 nm(λ_em);紫外检测波长设定:Kyn和IS为360 nm, Trp为302 nm。4种物质的回收率在91.62%~114.17%之间;线性范围分别为2.50~320.00 μ mol/L(Trp), 0.32~15.36 μ mol/L(Kyn), 3.27~104.60 nmol/L(5-Hiaa), 14.00~464.80 nmol/L(Kyna);检出限分别为0.078 μ mol/L(Trp), 0.056 μ mol/L(Kyn), 0.690 nmol/L(5-Hiaa), 1.290 nmol/L(Kyna)。利用该方法对30例正常孕妇和28例女性健康志愿者的血浆进行测定,结果表明两组间Trp, Kyn和Kyna含量有显著性差异。该方法操作简便,重复性好,灵敏度高,适合于临床检测。     

高效液相色谱-荧光检测法同时测定血清中的犬尿氨酸和犬尿喹啉酸

建立了在线衍生、双波长高效液相色谱-荧光检测器同时检测血清中犬尿氨酸(kynurenine, Kyn)和犬尿喹啉酸(kynurenic acid, KYNA)含量的方法。血清标本经5%高氯酸溶液去除蛋白质后,上清液直接进样分析测定。采用的色谱柱为Hypersil C8柱;流动相为0.25 mol/L醋酸锌-50 mmol/L醋酸溶液(含3%乙腈),流速为1.5 mL/min。在0～10 min时间段,在激发波长和发射波长分别为365 nm和480 nm时检测Kyn;10 min后,在激发波长和发射波长分别变换为344 nm和404 nm时检测KYNA。Kyn的保留时间约为8.1 min,线性范围为98～19600 nmol/L,最低检出浓度为50 nmol/L,平均回收率为94.88%,日内、日间测定值的相对标准偏差(RSD)均低于4%。KYNA的保留时间约为13.0 min,线性范围为2.62～1047 nmol/L,最低检出浓度为0.11 nmol/L,平均回收率为102.72%,日内、日间测定的RSD均低于4%。苯丙氨酸、酪氨酸、色氨酸和5-羟色胺等物质对目标物的检测无干扰。71例健康成人血清中,Kyn和KYNA含量分别为(1.40±0.34) μmol/L和(24.22±8.67) nmol/L。该方法简便、快速、灵敏、特异,适于临床和科研应用。     

普鲁士蓝增敏压电均相免疫分析法测定免疫球蛋白G

建立了普鲁士蓝增敏的均相压电免疫分析法,用于人尿液中免疫球蛋白G(IgG)的检测.本方法以蛋白质为种子,形成蛋白质-普鲁士蓝粒子,该粒子不断“滚雪球”增大,使得压电检测信号明显放大.在pH 4.0、盐浓度0.1 mol/L,5 mmol/L FeC13,2.5 mmol/L K4Fe(CN)6(K4Fe(CN)6与蛋白浓度比≥5000),FeCl3加样速度为0.5 μL/s的优化条件下,IgG的检测范围为0～625 nmol/L;检出限为2.4 nmol/L.α1-微球蛋白,β2-微球蛋白、尿微量白蛋白均无明显干扰.本方法用于临床样本的IgG测定,并与免疫比浊法进行对比,两者结果一致,表明本方法可行.     

Quantitative determination of dipyridamole in human plasma by high‐performance liquid chromatography–tandem mass spectrometry and its application to a pharmacokinetic study

Dipyridamole is a classic platelet inhibitor which has been a key medicine in clinical therapy of thrombosis and cerebrovascular disease. A rapid, selective and convenient method using high‐performance liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS) was developed for determination of dipyridamole in human plasma. After protein precipitation of 200 μL plasma with methanol, dipyridamole and diazepam (internal standard) were chromatographed on an Ultimate? XB‐C₁₈ (50 × 2.1 mm i.d, 3 μ) column with the mobile phase consisting of methanol–ammonium acetate (5 mM ; 80 : 20, v/v) at a flow rate of 0.25 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring mode via positive eletrospray ionization source (ESI⁺). The retention times of dipyridamole and diazepam were 1.4 and 1.2 min, respectively. The method was validated over a concentration range of 0.0180–4.50 μg/mL (r² ≥ 0.99) with a lower limit of quantitation (LLOQ) of 0.0180 μg/mL for dipyridamole. The intra‐ and inter‐day precisions (RSD) of the assay at all three QC levels were 1.6–12.7% with an accuracy (RE) of ?4.3–1.9%, which meets the requirements of the FDA guidance. The HPLC‐MS/MS method herein described was proved to be suitable for pharmacokinetic study of sustained‐release dipyridamole tablet in volunteers after oral administration. Copyright © 2009 John Wiley & Sons, Ltd.     

Determination of higher carboxylic acids in snow samples using solid-phase extraction and LC/MS-TOF

The objective of this work was to develop a method to determine the concentrations of higher organic acids in snow samples. The target species are the homologous aliphatic alpha,omega-dicarboxylic acids from C(5) to C(13), pinonic acid, pinic acid and phthalic acid. A preconcentration procedure utilizing solid phase extraction was developed and optimized using solutions of authentic standards. The influences of different parameters such as flow rate during extraction and the concentration of the eluent on the efficiency of the extraction procedure were investigated. The compounds of interest were separated by HPLC and detected by a quadrupole time-of-flight mass spectrometer (qTOF-MS). The recovery rate (extraction efficiency) of the extraction procedure was found to vary between 41% for tridecanedioic acid and 102% for adipic acid. The limits of detection were determined for all compounds and were between 0.9 nmol/L (dodecanedioic acid) and 29.5 nmol/L (pinonic acid). An exception is pinic acid, for which a considerably higher detection limit of 103.9 nmol/L was calculated. Snow samples were collected in December 2006 and January 2007 at the Fee glacier (Switzerland) from locations at heights from 3056 to 3580 m asl and from different depths within the snow layer. In total, the analysis of 61 single snow samples was performed, and the following compounds could be quantified: homologous aliphatic alpha,omega-dicarboxylic acids with 5-12 carbon atoms and phthalic acid. Tridecanedioic acid, pinonic and pinic acid were identified in the samples but were not quantified due to their low concentrations. The three most abundant acids found in the molten snow samples were glutaric acid (C(5)-di; 3.90 nmol/L), adipic acid (C(6)-di; 3.35 nmol/L) and phthalic acid (Ph; 3.04 nmol/L).     

高效液相色谱法测定水杨酸及其羟基化产物

 利用高效液相色谱法和荧光检测器建立了水杨酸及其主要的羟基化产物 2 ,3 二羟基苯甲酸和 2 ,5 二羟基苯甲酸的测定方法 ,并对水杨酸和H2 O2 光解反应体系进行了研究。     

高效液相色谱法测定环境水中超痕量双酚A

合成单分散双酚A(BPA)分子印迹微球并以其为高效液相色谱柱填充材料,建立了一种大体积直接进样HPLC法测定环境水中超痕量BPA的方法。方法的检出限(LOD)和测定限(LOQ)分别为0.03nmol/L和0.1nmol/L;在0.1~100nmol/L范围内具有良好的线性(r2=0.9983)。本方法用于表层湖水中BPA的检测,实际样品的加标回收率在99%~101.4%间;相对标准偏差(RSD%)低于8.1%(n=5)。     

Electrochemical determination of lead using bismuth modified glassy carbon electrode

Electrochemical application of bismuth film modified glassy carbon electrode was studied with the objective of lead detection. Bismuth film on glassy carbon substrate was formed in a plating solution of 2 mmol/L Bi(NO₃)₃, in 1 mol/L HCl at ?1.1 V (vs. Ag/AgCl) for 300 s. Lead was detected by differential pulse anodic voltammetry in acetate buffer of pH 5.0 in the concentration range of 7.5 nmol/L to 12.5 μmol/L. Factors influencing the anodic stripping performance, including deposition time, solution pH, Bi(III) concentration, potential, pulse amplitude, pulse width, have been optimized. Three linear calibration plots in the range 7.5 nmol/L to 0.1 μmol/L, 0.25 to 1 μmol/L, 2.5 to 12.5 μmol/L with regression coefficients of 0.991, 0.986 and 0.978 respectively were obtained. The theoretical detection limit equivalent to three times standard deviation for 7.5 nmol/L lead (n = 5) was calculated to be 5.25 nmol/L utilizing a 5 min deposition time and sensitivity 83.97 A L/mol. The sensitivity and detection limit of the method was compared with reported voltammetric methods for detection of lead and the result obtained was found to be promising for determination of lead.     

Amperometric biosensor based on D-aminoacid oxidase for the R-perindopril assay

A new amperometric biosensor based on D-aminoacid oxidase is described for the assay of R-perindopril. R-perindopril can be determined in the 400-20 nmol/L concentration range; the detection limit is 10 nmol/L. The selectivity was checked with S-perindopril, D- and L-proline, and polyvinylpyrrolidone. The main interfering species was D-proline. An automated system for the assay of R-perindopril based on the concept of flow injection with an amperometric biosensor (based on D-aminoacid oxidase) as detector is also described. The system is suitable for the on-line monitoring of R-perindopril at a sampling rate of 72 samples/h, in the linear range: 100 nmol/L -20 nmol/L with an RSD better than 0.09% (n = 10).     

蛋白质样品的非竞争性毛细管电泳免疫分析激光诱导荧光检测

以牛血清白蛋白与其单克隆抗体为样品,初步研究了蛋白质的非竞争性毛细管电泳免疫分析方法。混合温育可以在２０ｍｉｎ内达到平衡,电泳分离在１２ｍｉｎ内完成。当示踪物浓度为３２０ｎｍｏｌ／Ｌ时,所得到的标准曲线的线性范围为８￣１５０ｎｍｏｌ／Ｌ,检出限为５ｎｍｏｌ／Ｌ,并考察了缓冲溶液的浓度、ｐＨ、分离电压等因素的影响。     

Determination of traces of hemoglobin by square wave stripping voltammetry at a silver microelectrode

An electrochemical method was developed for the determination of traces of hemoglobin (Hb) by adsorption square wave voltammetric stripping at a bare silver microelectrode. Under optimum conditions the proposed method provided a linear response over the hemoglobin concentration range 5 to 100 nmol/L. The detection limit was 3 nmol/L. The relative standard deviation was 4.5% for 6 successive determinations at 50 nmol/L Hb. Other chemicals present in the sample did not interfere.     

The issue of HPLC determination of endogenous lipoic acid in human plasma

Lipoic acid (LA) is used extensively as a therapeutic agent for the treatment of various diseases. Many methods have been reported for the determination of LA plasma levels and its metabolites after its supplementation, but available information concerning endogenous plasma levels is still scarce. Studies which directly focused on determining the endogenous plasma levels provided highly controversial results, <4.9 nmol/L or 143.7–197.0 nmol/L. The main aim of this study was to verify the levels of free LA in the plasma of 40 individuals (17 women, 23 men). This group was nonsupplemented with LA and met the conditions for incorporation into the blood donors register. We measured the levels of LA using an HPLC method with very sensitive coulometric detection after previous sample preparation including deproteination and solid‐phase extraction with a Phenyl cartridge. Our limit of detection was 1.85 nmol/L and was better than the values reported in studies that directly focused on determining the endogenous plasma levels of LA: 2.4 and 4.9 nmol/L respectively. However, the levels of free LA in the plasma of nonsupplemented voluntary blood donors were not detectable in all cases. The presented results of our study show that endogenous concentrations of LA are <1.85 nmol/L.     

Selection of a DNA aptamer for the development of fluorescent aptasensor for carbaryl detection

An improved ssDNA library immobilized systematic evolution of ligands by enrichment(SELEX) was applied to select aptamers against carbaryl.After nine selection rounds,a highly enriched ssDNA pool was obtained.The Apta3 was demonstrated as the optimal aptame r.In order to facilitate the modification of aptamer,the Apta3 was further truncated with the dissociation constant(K_d) of 0.3 64 ± 0.055 μmol/L and a fluorescent aptasensor was developed.The linear range for carbaryl was from 100 nmol/L to1500 nmol/L,with the limit of detection was as low as 15.23 nmol/L.Besides,the biosensor was validated for the carbaryl spiked real samples,and the recoveries were between 97.7% and 107.3%.     

Quantitative determination of terbutaline in human plasma after administration of bambuterol using coupled columns and electrochemical detection

Summary A method is presented for quantitative determination of terbutaline after administration of its prodrug Bambuterol. Terbutaline is extracted from plasma by liquid-solid extraction on small C₁₈-cartridges. The extract is then analysed by coupled column liquid chromatography with amperometric detection. To inhibit the esterase catalyzed hydrolysis of bambuterol to terbutaline an analogue of bambuterol is added to the plasma sampling tubes. The within-day variation on spiked samples were 2.1% and 2.4% at 8.0 and 40.0 nmol/L respectively. The between-day variation on spiked samples (8.0 nmol/L) was 3.6% and on authentic samples 5.9% at the 11 nmol/L level. The absolute recovery was in the range 83–94% for terbutaline and the internal standard. The limit of quantitation was set at 4.0 nmol/L (C_v=3.4%, n=46). Subsidiary of AB Astra.     

Amperometric biosensor based on D-aminoacid oxidase for the R-perindopril assay

A new amperometric biosensor based on D-aminoacid oxidase is described for the assay of R-perindopril. R-perindopril can be determined in the 400–¶20 nmol/L concentration range; the detection limit is ¶10 nmol/L. The selectivity was checked with S-perindopril, D- and L-proline, and polyvinylpyrrolidone. The main interfering species was D-proline. An automated system for the assay of R-perindopril based on the concept of flow injection with an amperometric biosensor (based on D-aminoacid oxidase) as detector is also described. The system is suitable for the on-line monitoring of R-perindopril at a sampling rate of 72 samples/h, in the linear range: 100 nmol/L –20 nmol/L with an RSD better than 0.09% (n = 10).