Application of an LC–MS/MS Method for the Simultaneous Quantification of Homovanillic Acid and Vanillylmandelic Acid for the Diagnosis and Follow-Up of Neuroblastoma in 357 Patients

Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are end-stage metabolites of catecholamine and are clinical biomarkers for the diagnosis of neuroblastoma. For the first time in Korea, we implemented and validated a liquid chromatography tandem mass spectrometry (LC–MS/MS) assay to measure urinary concentrations of HVA and VMA according to Clinical and Laboratory Standards Institute guidelines. Our LC–MS/MS assay with minimal sample preparation was validated for linearity, lower limit of detection (LOD), lower limit of quantification (LLOQ), precision, accuracy, extraction recovery, carryover, matrix effect, and method comparison. A total of 1209 measurements was performed to measure HVA and VMA in spot urine between October 2019 and September 2020. The relationship between the two urinary markers, HVA and VMA, was analyzed and exhibited high agreement (89.1% agreement, kappa’s k = 0.6) and a strong correlation (Pearson’s r = 0.73). To our knowledge, this is the first study to utilize LC–MS/MS for simultaneous quantitation of spot urinary HVA and VMA and analyze the clinical application of both markers on a large scale for neuroblastoma patients.     

Monitoring Homovanillic Acid and Vanillylmandelic Acid in Human Urine by Capillary Electrophoresis with Electrochemical Detection

A simple,rapid and low-cost method of separation and determination of homovanillic acid and vanillylmandelic acid in human urine was developed based on capillary zone electrophoresis/amperometric detection with high sensitivity and good resolution.     

高效液相色谱－电化学检测法测定人尿液中的5－羟基吲哚乙酸和高香草酸

报道用反相高效液相色谱－电化学检测法同时测定人尿液中的５－羟基吲哚乙酸（５－ＨＩＡＡ）和高香草酸（ＨＶＡ），方法简便、快速、灵敏，尿样酸化后高心经稀释直接用于高效液相色谱（ＨＰＬＣ）分析。尿液中５－ＨＩＡＡ和ＨＶＡ的浓度在０．１２５～２０只μｇ／ｍＬ之间，回收率线性关系良好。测定了正常人和接触锰人群组尿液中的５－ＨＩＡＡ和ＨＶＡ。     

等速电泳法测定人齿菌斑培养液中乳酸含量

应用等速电泳法分离并测定了人齿菌斑培养液中的乳酸，测定回收率在９４．２％－１０２．２％之间，相对标准偏差小于４．５％，比较了抗龋者与易感龋者的菌斑在相同培养条件下的乳酸产量，并对不同饥饿时间下菌斑所产的乳酸进行了测定与比较。     

油田水中短链有机酸的等速电泳法研究

本文采用水相蒸发除去大量的氯离子,用等速电泳法对油田水中短链有机酸进行了定性定量分析。结果表明,油田地层水中有机酸成分主要有甲酸、乙酸、丙酸、丁酸、苯甲酸和乳酸等,本法回收率和相对标准偏差分别在96％～105％和2.4％～7.6％之间。     

自由溶液和筛分介质条件下芯片DNA瞬间等速电泳预浓缩的比较

利用芯片电泳方法考察瞬间等速电泳-筛分电泳偶联分析的结果,比较了自由溶液和筛分介质中DNA瞬间等速电泳的预浓缩效果.结果显示,相比较于筛分介质条件,自由溶液瞬间等速电泳有利于改善预浓缩和后续筛分电泳分离效果.对此结果的解释是:自由溶液条件下DNA迁移速度的提高可以延长瞬间等速电泳持续时间,有利于提高预浓缩效率.此外,样品压缩区带在自由溶液-筛分介质界面的二次富集也是预浓缩效果得到改善的原因之一.     

Measurement of Orotic Acid in Urine by Supercritical Fluid Chromatography

Orotic acid is an intermediate in the pathway of pyrimidine synthesis. It is an important substance but difficult to analyze in biological samples because it is present in trace amount in normal human urine, but greatly increased in inherited metabolic diseases and drug treatment. The methods used to determine orotic acid in urine were mainly high performance liquid chromatography (HPLC)1 and capillary zone electrophoresis (CZE)2. To date, the analysis of orotic acid in urine with SFC h…     

Determination of three Tumor Biomarkers (Homovanillic Acid,Vanillylmandelic Acid,and 5‐Hydroxyindole‐3‐Acetic Acid) Using Flow Injection Analysis with Amperometric Detection

Flow injection analysis with amperometric detection (FIA‐AD) at screen‐printed carbon electrodes (SPCEs) in optimum medium of Britton‐Robinson buffer (0.04 mol ? L^?1, pH 2.0) was used for the determination of three tumor biomarkers (homovanillic acid (HVA), vanillylmandelic acid (VMA), and 5‐hydroxyindole‐3‐acetic acid (5‐HIAA)). Dependences of the peak current on the concentration of biomarkers were linear in the whole tested concentration range from 0.05 to 100 μmol ? L^?1, with limits of detection (LODs) of 0.065 μmol ? L^?1 for HVA, 0.053 μmol ? L^?1 for VMA, and 0.033 μmol ? L^?1 for 5‐HIAA (calculated from peak heights), and 0.024 μmol ? L^?1 for HVA, 0.020 μmol ? L^?1 for VMA, and 0.012 μmol ? L^?1 for 5‐HIAA (calculated from peak areas), respectively.     

Determination of Phytic Acid in Urine by ICP Atomic Emission Spectrometry.

Abstract

A method to determine phytic acid within urine in the range 0.15–2 mg/l is reported. The method is based on the ICP atomic emission spectrometric determination of phosphorus, after previous separation and concentration of phytic acid using the anionic resin AG1-X8. The method has been applied successfully to determine phytic acid in synthetic urine and human urine.     

High Pressure Liquid Chromatographic Assay for Salicylic Acid and its Metabolites in Rat Urine

Abstract

An HPLC method for the determination of salicylic acid (SA), gentisic acid (GA), salicyluric acid (SU), and salicyl acyl glucuronide (SAG) in rat urine was developed. The method consisted of extracting SA, GA, and SU from acidified urine into 50:50 mixture of ethyl acetate and butyl chloride. Salicyl acyl glucuronide was extracted from neutral urine after conversion to salicyl hydroxamic acid with hydroxylamine. Salicyl phenolic glucuronide was estimated indirectly as the difference between total salicylate and sum of the four constituents mentioned above. Chromatographic separation was done on a C₁₈ column with U.V. detection at 310 nm using a mobile phase consisting of 5–10% acetonitrile in 3% glacial acetic acid. The extraction recovery of these compounds from spiked urine ranged from 90–108%. The detection limits were 10 μg/ml for GA, SU and SA, and 2.5 μg/ml for SHA. The method was applied to the study of salicylic acid metabolism in the rat.     

Micelle-Enhanced Spectrofluorimetric Method for the Determination of Antibacterial Trovafloxacin in Human Urine and Serum

A spectrofluorimetric method for the determination of trace amounts of the antibacterial trovafloxacin has been developed based on its native fluorescence in a micellar solution of sodium dodecyl sulfate (SDS). The wavelengths of excitation and emission were 270 nm and 410 nm, respectively. The optimised method allows the determination of 3.0–40.0 ng mL⁻¹ of trovafloxacin in 8 mM SDS solution and 0.1 M acetic acid-sodium acetate buffer solution (pH 5.5), with a relative standard deviation of 1.5% (for a level of 12.0 ng mL⁻¹) and a detection limit of 0.8 ng mL⁻¹. The method was applied to the determination of trovafloxacin in human urine and serum samples. It was validated using HPLC as a reference method. Recovery levels of the method reached 100% in all cases.     

Differential-Pulse Polarographic Determination of Some N-Substituted Phenothiazine Derivatives in Dosage Forms and Urine Through Treatment with Nitrous Acid

 A highly sensitive differential-pulse (DPP) polarographic method is described for the determination of three N-substituted phenothiazine derivatives, chlorpromazine (CZ), promazine (PZ) and promethazine (PMZ). The method involves the use of nitrous acid as an oxidant. Polarographically-active sulphoxides with diffusion-current constants (Id) of 2.53, 3.05 and 3.37 were obtained for CZ, PZ and PMZ, respectively. The polarographic waves were characterized as being diffusion-controlled, irreversible and partly affected by adsorption phenomena. All parameters affecting the oxidation process and polarographic behaviour were optimized and incorporated into the procedure. The limiting current-concentration plots in the DPP mode were rectilinear over the range: 0.006–0.1 mM, 0.005–0.08 mM and 0.008–0.1 mM for CZ, PZ and PMZ, respectively, with minimum detectability (S/N=3) of 3 × 10⁻⁷ M for CZ and PZ, and 4 × 10⁻⁷ M for PMZ, respectively. The kinetic parameters of the electrode reaction, including rate constant, free energy of activation ΔG and effect of temperature on both parameters were studied. The proposed method was successfully applied to the determination of phenothiazines in dosage forms; the results obtained were in agreement with those given with the official methods. The method was further applied to the determination of promazine in spiked human urine. The percentage recovery was 96.86 ± 0.30. The advantages of the proposed method over other reported methods were discussed. A proposal of the electrode reaction was made. Received June 1, 1999. Revision March 10, 2000.     

毛细管区带电泳法测定尿中甲酸含量

采用高效毛细管区带电泳法测定人体尿中甲酸含量,尿样经过滤后直接进样,方法简单、快速,测定结果令人满意。电泳电解质体系采用５ｍｍｏｌ／Ｌ邻苯二甲酸氢钾,０．５ｍｍｏｌ／Ｌ十六烷基三甲基溴化铵,ｐＨ６,５０ｃｍ×５０μｍｉ．ｄ．熔融石英毛细管（有效长度４８．５ｃｍ）,检测波长２１０ｎｍ,负电源,分离电压３０ｋＶ,压力进样,恒温２５℃,每次电泳前用０．１ｍｏｌ／ＬＮａＯＨ及缓冲溶液对毛细管各冲洗５ｍｉｎ。同时,采用检测波长与参比波长对调的方法使负峰转变为正峰。     

抗坏血酸存在下离子色谱法直接测定尿液中草酸含量的研究

建立了抗坏血酸存在下离子色谱法电导检测尿液中草酸的方法。样品经稀释后用0.22μm滤膜过滤,直接进样,以15 mmol·L-1Na HCO3(p H 8.3)为淋洗液,用Ion Pac AS12A阴离子分离柱(200 mm×4mm)分离,电导检测器进行检测。实验结果表明,草酸与其他常见阴离子在16 min内能够很好地分离,在此条件下抗坏血酸能够在80 min内保持稳定,不会降解为草酸而影响测量结果。测试条件下,草酸在0.2~40mg·L-1浓度范围内呈良好的线性关系,相关系数(r)为0.999 9,检出限达0.02 mg·L-1,加标回收率为96.9%~103.3%,可用于实际样品的测试。     

吸附溶出伏安法测定血浆和尿液中的尿酸

在Ｂｒｉｔｔｏｎ－Ｒｏｂｉｎｓｏｎ缓冲体系中，研究了尿酸的伏安行为，实验表明，尿酸在悬汞电极上具有吸附行为，并于－０．０８Ｖ产生灵敏的峰电流。该法可用于尿液和血浆中尿酸的测定，回收率为９５－１０５％。     

Simultaneous Determination of Cefepime and the Quinolones Garenoxacin, Moxifloxacin and Levofloxacin in Human Urine by HPLC-UV

A liquid chromatographic method with a C₁₈ column and acetonitrile/0.1 M phosphoric acid/ sodium hydroxide buffer (pH 3.0)/0.01 M n-octylamine (pH 3.0) as mobile phase in gradient mode has been developed and optimised for the simultaneous determination of the cephalosporin cefepime and the quinolones garenoxacin, levofloxacin and moxifloxacin. Identification and quantification was carried out with a diode-array UV detector, with working wavelengths of 256 nm for cefepime, 292 nm for levofloxacin, 294 nm for moxifloxacin and 282 nm for garenoxacin. The mobile flow-rate and sample volume injected were 1 mL min⁻¹ and 20 µL, respectively. The retention times and detection limits for each antibiotic were 4.9 min and 1.9 µg mL⁻¹ for cefepime, 7.5 min and 2.2 µg mL⁻¹ for levofloxacin, 8.9 min and 2.7 µg mL⁻¹ for moxifloxacin and 10.7 min and 1.8 µg mL⁻¹ for garenoxacin, respectively. The method was applied to the determination of the four molecules in spiked samples of human urine.     

羟基吡啶功能化螺吡喃探针检测尿样中柠檬酸的研究

柠檬酸是多种疾病潜在的生物标记物,因此发展对柠檬酸灵敏、专一的快速检测方法势在必行。该文设计合成了一种羟基吡啶功能化水溶性螺吡喃探针,通过在螺吡喃骨架上引入羟基吡啶基团,利用螺吡喃螺碳及羟基的协同作用增强了探针对柠檬酸的结合能力。结果表明,该探针对柠檬酸有选择性响应,并伴随着明显的颜色变化及荧光猝灭。据此建立了尿样中柠檬酸的荧光检测方法,方法线性范围为20～120μmol/L,检出限为1.0μmol/L。人工尿液中柠檬酸的回收率为85.7%～92.2%,相对标准偏差为3.0%～9.2%。该方法具有操作简便、准确及选择性好的优点,在尿样柠檬酸检测中具有良好的应用潜力。     

液相色谱-串联质谱法测定尿液中的泛酸

该文建立了检测尿液中泛酸含量的液相色谱-串联质谱(HPLC-MS/MS)方法,尿液经过离心、稀释后,采用ACPUITY UPLC SS T3(2.1 mm×100 mm,1.8μm)色谱柱进行分离,电喷雾正离子模式电离,多反应监测模式进行检测,方法的线性关系良好(r=0.999 3),方法检出限为0.46 ng/m L,回收率为87.9%~95.3%,相对标准偏差(RSD)为2.5%~13.0%。该方法具有灵敏度高、分析时间短等特点,可用于尿液中泛酸含量的分析。     

Determination of 4-Hydroxyphenyllactic Acid in Human Urine by Magnetic Solid-Phase Extraction and High-Performance Anion-Exchange Chromatography with Fluorescence Detection

A novel, simple, rapid, and accurate method is reported for the determination of 4-hydroxyphenyllactic acid in human urine by high-performance anion-exchange chromatography with fluorescence detection and magnetic solid-phase extraction. The separation and pretreatment conditions for urine were optimized. The isolation of 4-hydroxyphenyllactic acid was performed with isocratic elution with 4?mmol?L^?1 sodium hydroxide at 0.45?mL min^?1. Fluorescence detection was performed at an excitation wavelength of 277?nm and an emission wavelength of 340?nm. Under the optimized conditions, the linear dynamic range and the limit of detection for 4-hydroxyphenyllactic acid were 0.05–10 and 0.020?mg?L^?1, respectively. The recovery for the analyte was from 86.5 to 105.5%, with relative standard derivations less than 4.12%. The method was used for the determination of 4-hydroxyphenyllactic acid in human urine. Statistically significant differences in the 4-hydroxyphenyllactic acid concentration in urine were obtained between healthy control and individuals with breast cancer.     

A Semi-Automated Colorimetric Method for the Analysis of Dihydroxyphenylacetic Acid (DOPAC) in Urine of Parkinsonian Patients Receiving L-Dopa

Abstract

An analytical system is presented which permits the separation and analysis of urinary dihydroxyphenylacetic acid (DOPAC). This method is suitable for analyzing the urine from both normal and Parkinsonian patients receiving L-Dopa.