Identification and quantification of urinary benzo[a]pyrene and its metabolites from asphalt fume exposed mice by microflow LC coupled to hybrid quadrupole time-of-flight mass spectrometry

Prolonged, extensive exposure to asphalt fume has been associated with several adverse health effects. Inhaled polycyclic aromatic hydrocarbons (PAHs) from asphalt fume exposure have been suspected of inducing such effects. In this study, a bioanalytical method was proposed and evaluated to identify and quantify benzo[a]pyrene and its hydroxy-metabolites. This method is based on coupling a microflow liquid chromatography (LC) to a hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometry (Q-TOFMS). In the experiment, thirty-two B6C3FI mice were exposed to asphalt fume in a whole body inhalation chamber for 10 days (4 h day(-1)) and twelve other mice were used as controls. The asphalt fume was generated at 180 degrees C and the concentrations in the animal exposure chamber ranged 175-182 mg m(-3). Benzo[a]pyrene and its metabolites of 3-hydroxybenzo[a]pyrene, benzo[a]pyrene-7,8-dihydrodiol(+/-), benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide(+/-), and benzo[a]pyrene-7,8,9,10-tetrahydrotetrol(+/-) in the urine of asphalt fume exposed mice were identified and found at 3.18 ng 100 mL(-1), 31.36 ng 100 mL(-1), 11.56 ng 100 mL(-1), 54.92 ng 100 mL(-1), and 45.23 ng 100 mL(-1) respectively. The results revealed that the urinary benzo[a]pyrene and its hydroxy-metabolites from exposed mice were at significantly higher levels (p < 0.001) than those from the control groups. Compared with several other technologies such as HPLC-UV and HPLC-fluorescence, the new method is more sensitive and selective, and it can also provide additional useful information on the structures of the metabolites. Hence, this method can be used to perform the assessment and to study the mechanisms of the adverse health effects. The fragmentation patterns established in this study can also be used to identify and quantify PAH metabolites in other biological fluids.     

Highly sensitive routine method for urinary 3-hydroxybenzo[a]pyrene quantitation using liquid chromatography-fluorescence detection and automated off-line solid phase extraction

Many workers and also the general population are exposed to polycyclic aromatic hydrocarbons (PAHs), and benzo[a]pyrene (BaP) was recently classified as carcinogenic for humans (group 1) by the International Agency for Research on Cancer. Biomonitoring of PAHs exposure is usually performed by urinary 1-hydroxypyrene (1-OHP) analysis. 1-OHP is a metabolite of pyrene, a non-carcinogenic PAH. In this work, we developed a very simple but highly sensitive analytical method of quantifying one urinary metabolite of BaP, 3-hydroxybenzo[a]pyrene (3-OHBaP), to evaluate carcinogenic PAHs exposure. After hydrolysis of 10 mL urine for two hours and concentration by automated off-line solid phase extraction, the sample was injected in a column-switching high-performance liquid chromatography fluorescence detection system. The limit of quantification was 0.2 pmol L(-1) (0.05 ng L(-1)) and the limit of detection was estimated at 0.07 pmol L(-1) (0.02 ng L(-1)). Linearity was established for 3-OHBaP concentrations ranging from 0.4 to 74.5 pmol L(-1) (0.1 to 20 ng L(-1)). Relative within-day standard deviation was less than 3% and relative between-day standard deviation was less than 4%. In non-occupationally exposed subjects, median concentrations for smokers compared with non-smokers were 3.5 times higher for 1-OHP (p<0.001) and 2 times higher for 3-OHBaP (p<0.05). The two urinary biomarkers were correlated in smokers (ρ=0.636; p<0.05; n=10) but not in non-smokers (ρ=0.09; p>0.05; n=21).     

Determination of polycyclic aromatic hydrocarbons with molecular weight 302 in water samples by solid-phase nano-extraction and laser excited time-resolved Shpol'skii spectroscopy

Monitoring of high-molecular weight polycyclic aromatic hydrocarbons (HMW-PAH) via simple and cost effective methods still remains a challenge. In this article, we combine solid-phase nano-extraction (SPNE) and 4.2 K laser-excited time resolved Shpol'skii spectroscopy (LETRSS) into a valuable alternative for the water analysis of dibenzo[a,l]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene and naphtho[2,3-a]pyrene. In comparison to the original SPNE procedure, the present method improves PAH recoveries and reduces extraction time from 30 to 20 min per sample. Quantitative release of HMW-PAH into the Shpol'skii matrix (n-octane) is best accomplished with a mixture of 48 μL of methanol and 2 μL of 1-pentanethiol. Their migration into the 50 μL layer of n-octane provides highly resolved spectra with distinct fluorescence lifetimes for unambiguous isomer determination. Complete analysis takes less than 30 min per sample and consumes only 100 micro-liters of organic solvents. 500 μL of water are sufficient to obtain limits of detection ranging from 16 ng L(-1) (dibenzo[a,l]pyrene) to 55 ng L(-1) (dibenzo[a,i]pyrene), relative standard deviations better than 3% and analytical recoveries above 90%. Although a straightforward comparison to chromatographic methods is not possible because of the lack of analytical figures of merit on HMW-PAH, the excellent precision of measurements, limits of detection and overall recoveries makes SPNE-LETRSS an attractive approach to water analysis of HMW-PAH.     

三聚氰胺单试剂荧光偏振免疫分析法

用三聚氰胺半抗原(MEL)与异硫氰酸荧光素乙二胺(EDF)反应合成了荧光示踪物MEL-EDF, 示踪物与三聚氰胺抗体反应得到稳定的单试剂免疫复合物, 考察了其结合动力学和置换动力学过程, 建立了一种三聚氰胺单试剂荧光偏振免疫分析方法(SR-FPIA). 该方法的检出限(LOD)为3.2 ng/mL, 半抑制浓度(IC₅₀)为157.1 ng/mL, 检测范围(IC₂₀~IC₈₀)为12.2~1012.4 ng/mL, 能够满足国家标准和国际食品法典委员会(CAC)对于食品中三聚氰胺最高残留限量的检测要求. 整个检测过程在15 min内完成, 并且单试剂免疫复合物在4℃下至少可稳定保存30 d以上. 本研究对开发三聚氰胺新型免疫检测方法具有重要意义.     

高效液相色谱法测定生活饮用水中的苯并[a]芘

建立高效液相色谱二极管阵列检测器检测生活饮用水苯并[a]芘的测定方法。采用C18反相色谱柱(150mm×4.6 mm,5μm),在流动相为甲醇–水(体积比为90∶10)、流量1.0 mL/min、检测波长295 nm、柱温35℃、进样体积20μL的条件下测定生活饮用水中苯并[a]芘。该方法检出限为6 ng/L,线性范围0～100 ng/mL,加标回收率为88.1%～93.4%,测定结果的相对标准偏差为1.06%(n=9)。该法样品预处理简单,分离度高,分析时间短,适用于生活饮用水中苯并[a]芘的准确定性定量测定。     

Single-laboratory validation of a gas chromatography-mass spectrometry method for quantitation of 15 European priority polycyclic aromatic hydrocarbons in spiked smoke flavourings

An existing method was adapted to the purpose and validated in-house according to the IUPAC harmonised guideline for the determination of 15 EU priority polycyclic aromatic hydrocarbons (PAHs) in primary smoke condensates (PSCs) that are used to produce smoke flavourings for human consumption. Limits of detection (LOD) varied between 0.1 and 1.3 microg/kg, limits of quantitation (LOQ) between 0.5 and 4 microg/kg for the various PAHs in PSCs. The coefficient of variance of the repeatability was between 0.7% (benzo[a]pyrene) and 30% (dibenzo[a,h]pyrene) relative standard deviation, depending on the analyte. The recoveries varied between 100 and 102% (dibenzo[a,l]pyrene) and 69-83% (dibenzo[a,h]pyrene) over the analytical range of 5-35 microg/kg.     

A sensitive column-switching HPLC method for aripiprazole and dehydroaripiprazole and its application to human pharmacokinetic studies

A simple and sensitive column‐switching HPLC‐UV method was developed for the simultaneous determination of aripiprazole, a novel atypical antipsychotic drug, and its active metabolite, dehydroaripiprazole in human plasma. Aripiprazole, its active metabolite and 7‐[5‐[4‐(3‐chloro‐2‐methylphenyl)‐1‐piperazinyl]pentyloxy]‐3,4‐dihydro‐2(1H)‐quinolinone (OPC‐14558) as an internal standard were extracted from 1 mL of plasma using a mixture of chloroform/n‐heptane (3:7, v/v), and the extract was injected into a column I (TSK BSA‐ODS/S precolumn, 5 μm) for cleanup and column II (C₁₈ STR ODS‐II analytical column, 5 μm) for separation. Peaks were detected with an UV detector set at a wavelength of 254 nm, and the total time for chromatographic separation was ～20 min. Mean absolute recoveries were 74.0 and 74.7% for aripiprazole and dehydroaripiprazole, respectively. Intra‐ and inter‐day CVs were less than 7.5 and 7.1% for aripiprazole concentrations ranging from 2 to 600 ng/mL, and 9.2 and 4.5% for dehydroaripiprazole concentrations ranging from 2 to 160 ng/mL. The validated concentration ranges for this method were 1–500 ng/mL and the limits of detection were 0.5 ng/mL for both aripiprazole and dehydroaripiprazole. This method was applied to pharmacokinetic study in human volunteers and patients taking aripiprazole.     

A New and Concise Synthesis of 3-Hydroxybenzo[c]phenanthrene and 12-Hydroxybenzo[g]chrysene, Useful Intermediates for the Synthesis of Fjord-Region Diol Epoxides of Benzo[c]phenanthrene and Benzo[g]chrysene

A new strategy which involves a palladium-catalyzed cross-coupling reaction has been developed for the rapid synthesis of 3-hydroxybenzo[c]phenanthrene (5) and 12-hydroxybenzo[g]chrysene (6). These phenolic compounds are the key intermediates for the synthesis of highly carcinogenic fjord-region diol epoxide metabolites 3 and 4 of benzo[c]phenanthrene (1) and benzo[g]chrysene (2). The cross-coupling reaction of 2-bromo-5-methoxybenzaldehyde (9) with naphthalene-1-boronic acid (7) and phenanthrene-9-boronic acid (8) produced 2-(1-naphthyl)-5-methoxybenzaldehyde (10) and 2-(9-phenanthryl)-5-methoxybenzaldehyde (11), respectively, in quantitative yields. After reaction of these aldehydes with trimethylsulfonium iodide under phase-transfer conditions or with the Wittig reagent obtained from (methoxymethyl)triphenylphosphonium bromide and phenyllithium to generate an oxiranyl or methoxyethene side chain, the acid-catalyzed cyclization with methanesulfonic acid (or boron trifluoride) produced 3-methoxybenzo[c]phenanthrene (16) and 12-methoxybenzo[g]chrysene (17) in 61-64% yields. Finally, demethylation of these methoxy derivatives 16 and 17 with boron tribromide resulted in the formation of the hydroxy analogues 5 and 6, respectively. The availability of this short and high-yielding regiospecific method for the synthesis of phenols 5 and 6 should allow the preparative-scale synthesis of the fjord-region diol epoxides 3 and 4. These diol epoxides are required as starting compounds for the synthesis of site-specifically modified oligonucleotides which are critically needed to elucidate the mechanism of carcinogenesis at the molecular level.     

Quantification of atrazine and its metabolites in urine by on-line solid-phase extraction–high-performance liquid chromatography–tandem mass spectrometry

We have developed a method using on-line solid-phase extraction–high-performance liquid chromatography–tandem mass spectrometry (SPE-HPLC-MS/MS) and isotope dilution quantification to measure atrazine and seven atrazine metabolites in urine. The metabolites measured were hydroxyatrazine, diaminochloroatrazine, desisopropylatrazine, desethylatrazine, desethylatrazine mercapturate, atrazine mercaturate and atrazine itself. Our method has good precision (relative standard deviations ranging from 4 to 20% at 5, 10 and 50 ng/mL), extraction efficiencies of 67 to 102% at 5 and 25 ng/mL, relative recoveries of 87 to 112% at 5, 25, 50 and 100 ng/mL limits of detection (LOD) ranging from 0.03 to 2.80 ng/mL. The linear range of our method spans from the analyte LOD to 100 ng/mL (40 ng/mL for atrazine and atrazine mercapturate) with R ² values of greater than 0.999 and errors about the slope of less than 3%. Our method is rapid, cost-effective and suitable for large-scale sample analyses and is easily adaptable to other biological matrices. More importantly, this method will allow us to better assess human exposure to atrazine-related chemicals. Figure A schematic representation showing the elution of the analytes from the solid-phase extraction cartridge onto the analytical column for chromatographic separation prior to MS/MS analysis     

高灵敏甲胎蛋白夹心免疫传感器的制备

构建了新型甲胎蛋白(AFP)夹心免疫传感器.采用金纳米粒子-氧化石墨烯-普鲁士蓝纳米立方体(AuNP-GO-PBNCs)纳米复合材料标记甲胎蛋白(AFP)二抗,将制备的金-聚多巴胺-四氧化三铁(Au-PDA-Fe3O4)磁性纳米复合物固定在自制的磁性电极表面,通过吸附作用固定AFP一抗,用牛血清白蛋白(BSA)封闭电极上的非特异性吸附位点.在37℃下与AFP抗原溶液孵育50 min,最后将电极放入AuNP-GO-PBNCs纳米复合材料标记的二抗溶液中孵育,基于此建立了采用普鲁士蓝(PB)标记的的夹心免疫传感器检测AFP的方法.在最佳实验条件下,PB催化H2O2氧化的响应电流与AFP的浓度表现出两段线性关系,线性范围分别为0.005~1.000 ng/mL和1~20 ng/mL, 检出限(LOD, S/N=3)为1.0 pg/mL.本方法具有灵敏度高、选择性好的特点.     

固相萃取-液相色谱-串联质谱法同时测定尿液中9种多环芳烃代谢物

建立了固相萃取-液相色谱-串联质谱同时测定尿中2-羟基萘、1-羟基萘、2-羟基芴、3-羟基菲、1-羟基芘等9种多环芳烃代谢物的液相色谱-串联质谱测定方法。尿样中结合态的多环芳烃代谢物在β-葡萄糖苷酸酶-芳基硫酸酯酶缓冲液(pH 5.0)作用下,于37℃水浴中避光水解4 h后,以C18固相萃取小柱富集、净化,以甲醇洗脱,采用Waters Symmetry C18色谱柱,流动相为乙腈-0.2%氨水(72∶27,V/V)等度淋洗分离后进入质谱测定。在喷雾电压4 kV,毛细管温度300℃下,以3-羟基菲13C为内标,采用SRM模式负离子扫描方式测定,内标法定量。9种多环芳烃代谢物在尿中的线性范围为0.90～100μg/L;相关系数为0.9970～0.9990;回收率为79.0%～119.8%;相对标准偏差为4.3%～12.4%;检出限为0.04～0.90μg/L;结果表明,本方法可用于尿中9种多环芳烃代谢物的测定。     

Monitoring of biogenic amines and drugs of various therapeutic groups in urine samples with use of HPLC

A high‐performance liquid chromatography method for simultaneous separation and determination of biogenic amines [dopamine, epinephrine, serotonin and its six metabolites (normetanephrine, metanephrine, 3,4‐dihydroxyphenylacetic acid, 4‐hydroxy‐3‐methoxyphenylglycol, homovanilic acid and 5‐hydroxyindoloacetic acid)] with drugs from different therapeutically groups [analgesics (paracetamol, metamizol), diuretics (furosemide) and antibiotics (cefazolin, fluconazole)] was developed. A chromatographic column with pre‐column with octadecylsilane phase (C_18e) and two detectors – diode array serial connected and fluorescence – was used. Gradient elution of mixture of acetate buffer (pH 4.66) and methanol as a mobile phase was applied. The limit of detection (LOD) of 8–10 ng/mL and limit of quantitation (LOQ) of 24–30 ng/mL for biogenic amines, as well as the LOD of 50–100 ng/mL and the LOQ of 150–300 ng/mL for drugs, were determined. The applied sample preparation method allowed recoveries of 93% for the biogenic amines and 92% for the drugs to be achieved. The developed procedure has been applied to simultaneous determination of the examined compounds in urine samples and could be used in clinical analysis. Copyright © 2015 John Wiley & Sons, Ltd.     

中性解吸电喷雾萃取电离质谱法直接检测蜂蜜中的敌敌畏

采用中性解吸电喷雾萃取电离质谱( ND-EESI-MS)技术,在无需样品预处理的条件下,建立了对蜂蜜中敌敌畏直接快速检测的方法。在正离子模式下,敌敌畏质子化离子峰位于 m/z 223,二级特征离子为m/z 109和127。在优化的条件下,以m/z 127的信号强度为定量指标,建立了蜂蜜中敌敌畏残留的定量检测方法。结果表明,在蜂蜜基质中,敌敌畏在5~1000 ng/mL浓度范围内与m/z 127的信号强度线性关系良好,相关系数为0.998,检出限为1.0 ng/mL(S/N=3);蜂蜜中3个加标水平(10,30和400 ng/mL)的敌敌畏的回收率为93.0％~103.0％,精密度(RSDs)小于4.4％。同时采用气相色谱(火焰光度检测器)方法作为对照方法,检测敌敌畏加标蜂蜜样品,结果表明,加标蜂蜜在5~1000 ng/mL浓度范围内与峰面积线性关系良好,相关系数为0.999,检出限为1.6 ng/mL;10,30和400 ng/mL 3个水平加标蜂蜜的回收率为94.9％~110.3％,精密度小于7.6％。     

A luminescence immunoassay test method for determining benzo[a]pyrene in natural water

A test method is developed for determining benzo[a]pyrene in natural water, based on the use of a polyethylene filter (frit) with adsorbed specific antibodies, placed within a transparent column. In passing a test solution, the analyte is adsorbed on the frit similarly to the process implemented in immunoaffinity preconcentration. The added conjugate of a labeled analyte takes the remained vacant binding sites of antibodies. Luminescent semiconductor nanoparticles (quantum dots) CdSe/ZnS, used as labels, enable visual determination under irradiation with UV light. The limit of detection for benzo[a]pyrene in water is ~0.5 ng/mL.     

Application of stir bar sorptive extraction to the determination of polycyclic aromatic hydrocarbons in aqueous samples

The technique of stir bar sorptive extraction is used for the determination of polycyclic aromatic hydrocarbons (PAH) in aqueous samples. The PAHs are extracted with 10-mm stir bars (Gerstel Twister) coated with 0.5 mm polydimethylsiloxane and analyzed with a gas chromatography-mass spectrometry system. The influence of methanol and hyamine addition to the samples for preventing wall effects is investigated at 100 ng/l. The results indicate improved sensitivity using hyamine addition to the samples. The optimal extraction time was found to be between 3 and 4 h. The reproducibility of the method, as determined by nine replicate measurements, is between 5 and 15% at 10 ng/l and between 3 and 9% at 50 ng/l. Carry-over, which was evaluated at 500 ng/l by desorbing the same Twister three times, seems to be negligible for most of the compounds. In worst cases, carry-over of up to 7% was found for indeno[1,2,3]pyrene, dibenz[a,h]anthracene, and benzo[g,h,i]perylene. The technique shows excellent linearities for 5 point calibrations. Detection limits are between 0.1 and 2 ng/l.     

A novel fast method for aqueous derivatization of THC,OH‐THC and THC‐COOH in human whole blood and urine samples for routine forensic analyses

A novel aqueous in situ derivatization procedure with propyl chloroformate (PCF) for the simultaneous, quantitative analysis of Δ⁹‐tetrahydrocannabinol (THC), 11‐hydroxy‐Δ⁹‐tetrahydrocannabinol (OH‐THC) and 11‐nor‐Δ⁹‐tetrahydrocannabinol‐carboxylic acid (THC‐COOH) in human blood and urine is proposed. Unlike current methods based on the silylating agent [N,O‐bis(trimethylsilyl)trifluoroacetamide] added in an anhydrous environment, this new proposed method allows the addition of the derivatizing agent (propyl chloroformate, PCF) directly to the deproteinized blood and recovery of the derivatives by liquid–liquid extraction. This novel method can be also used for hydrolyzed urine samples. It is faster than the traditional method involving a derivatization with trimethyloxonium tetrafluoroborate. The analytes are separated, detected and quantified by gas chromatography–mass spectrometry in selected ion monitoring mode (SIM). The method was validated in terms of selectivity, capacity of identification, limits of detection (LOD) and quantification (LOQ), carryover, linearity, intra‐assay precision, inter‐assay precision and accuracy. The LOD and LOQ in hydrolyzed urine were 0.5 and 1.3 ng/mL for THC and 1.2 and 2.6 ng/mL for THC‐COOH, respectively. In blood, the LOD and LOQ were 0.2 and 0.5 ng/mL for THC, 0.2 and 0.6 ng/mL for OH‐THC, and 0.9 and 2.4 ng/mL for THC‐COOH, respectively. This method was applied to 35 urine samples and 50 blood samples resulting to be equivalent to the previously used ones with the advantage of a simpler method and faster sample processing time. We believe that this method will be a more convenient option for the routine analysis of cannabinoids in toxicological and forensic laboratories.     

Determination of rapamycin: quantification of the sodiated species by an ion trap mass spectrometer as an alternative to the ammoniated complex analysis by triple quadrupole

Rapamycin is a potent immunosuppressive drug capable of significantly reducing acute graft rejection in kidney, liver and heart transplant patients. Its immunosuppressive activity and adverse effects have been related to rapamycin concentration, and therapeutic drug monitoring of the drug is deemed appropriate. This work was aimed at developing a new quantification method based on the isolation of the [M+Na]+ ion as precursor and its further fragmentation through an ion trap mass spectrometer equipped with an electrospray ionization source. A limit of detection (LOD) of 0.7 ng/mL was obtained, while the lower limit of quantification (LLOQ) was 2.4 ng/mL. The accuracy and reproducibility of the responses were evaluated and compared with results obtained when the [M+NH4]+ ion was chosen as the precursor in a triple quadrupole mass spectrometer. In this case the LOD was 0.5 ng/mL and the LLOQ 1.7 ng/mL. Data showed that it would be possible to use the quantification of the sodiated species for the routine determination of rapamycin, as an alternative to the commonly adopted method based on the ammoniated complex.     

RSR13, a potential athletic performance enhancement agent: detection in urine by gas chromatography/mass spectrometry.

RSR13 (2-[4-[[(3,5-dimethylanilino)carbonyl]methyl]phenoxyl]-2-methylpropionic acid) is a synthetic allosteric modifier of hemoglobin that is currently in a phase III clinical trial as a radio-enhancing agent. RSR13 has been shown to increase maximum oxygen uptake (VO(2max)) in a canine skeletal model, which makes it a potential performance-enhancing agent for endurance athletes, since VO(2max) is an index of aerobic capacity. In this study we present a method for the detection of RSR13-bis-TMS in human urine by gas chromatography/electron impact ionization mass spectrometry (GC/EI-MS) suitable for doping control laboratories. The presence of RSR13 is detected by monitoring the ions m/z 485 ([M](+.)) and 470 ([M - CH3](+)). The limit of detection (LOD) is less than 2 ng/mL in urine. Urine samples collected from clinical trial subjects immediately prior to receiving an infusion of RSR13 showed no evidence of RSR13, whereas post-infusion urine samples contained up to 1181 microg/mL. A urine sample collected 36 h after administration of a small dose (10 mg/kg) and diluted 100-fold showed a signal 80 times higher than the LOD. Urine samples obtained from 100 randomly selected athletes in our routine testing program did not show any traces of RSR13. Sport authorities may wish to add RSR13 to the list of prohibited substances.     

Multivariate calibration applied to synchronous fluorescence spectrometry. Simultaneous determination of polycyclic aromatic hydrocarbons in water samples

Synchronous fluorescence spectra of mixtures containing ten polycyclic aromatic hydrocarbons (anthracene, benz[a]anthracene, benzo[a]pyrene, chrysene, fluoranthene, fluorene, naphthalene, perylene, phenanthrene and pyrene) have been used for the determination of these compounds by Partial Least Squares Regression (PLSR), using both PLS-1 and PLS-2. Different procedures have been used for the pretreatment of the data in order to obtain better models, and the size of the calibration matrix has also been studied. The best models have been used for the determination of the above mentioned PAHs in spiked natural water samples at concentration levels between 4 and 20 ng ml⁻¹. Recoveries ranged from 80 to 120% in most cases, although fluorene gave significantly lower results.     

Determination of levodopa and biogenic amines in urine samples using high-performance liquid chromatography

A chromatographic system is developed for the separation and determination of levodopa, biogenic amines, and their metabolites from the catecholamines group: dopamine (DA), epinephrine (E), normetanephrine (NMN), metanephrine (MN), 3,4-dihydroxyphenylacetic acid (DOMA), 3-metoxy-4-hydroxyphenyl-glycol (MHPG), and homovanillic acid (HVA); and indoloamines group: serotonin (5HT) and 5-hydroxyindole-3-acetic acid (5HIAA) in urine. The limit of detection (LOD) and limit of quantitation (LOQ) are determined for all compounds with signal-to-noise ratio (S/N) of 3 and 10, respectively. LOD 10 (ng/mL) and LOQ 30 (ng/mL) are determined for L-DOPA, DOMA, E, NMN, DA, MN, and MHPG, as well as LOD 8 (ng/mL) and LOQ 24 (ng/mL) for HVA, 5HT, and 5HIAA. A fluorescence detector is used. Gradient elution with acetate buffer (pH=4.66) with methanol is applied. In urine samples from patients treated with levodopa, the following concentrations (microg/mL) of analytes are determined: L-DOPA 3.73-46.80, DOMA 1.43-28.43, E 0.83-13.57, NMN 2.58-8.81, DA 24.07-62.11, MN 0.89-66.20, MHPG 6.72-63.64, 5HT 22.96-95.27, 5HIAA 1.45-14.77, and HVA 0.21-15.07.