Simultaneous measurement of volatile sulfur compounds using ascorbic acid for oxidant removal and gas chromatography-flame photometric detection

The use of Eriochrome Black T in an alkaline, 40% methanol solution was found to be appropriate as post-column reagent for the determination of rare earths by ion chromatography. Detection of individual lanthanides and lanthanum was carried out at 512 nm and 650 nm after separation by dynamic cation exchange chromatography with gradient elution on C18 column and employing a solution containing alpha-hydroxyisobutiric acid/sodium octanesulfonate at pH 3.8 as eluent. The effect of the presence of micelles in the post-column reagent was studied. Sensitivities obtained by the addition of the cationic surfactants cetylpyridinium chloride (CPC) and hexadecyltrimethylammonium bromide (CTAB) were lower than those measured without surfactant addition. In some cases, the signal was totally suppressed. No change in sensitivity was observed with non-ionic (Triton X-100) or anionic (sodium dodecylsulphate, SDS) surfactants but a slight improvement in the baseline noise was observed with the SDS. An evaluation of the influence of chemical and operational variables on the post column reaction (PCR) reagent was carried out either by spectrophotometric tests or by chromatographic experiments. A comparison was performed between three PCR reagents: Eriochrome Black T and xylenol orange in the presence of a cationic surfactant and arsenazo III. Calibration response was linear up to an analyte concentration of 5.0 micrograms ml-1. Absolute detection limits lower than 7 and 17 ng were obtained at the detection wavelengths of 650 nm and 512 nm respectively, for all the natural lanthanides and lanthanum.     

Determination of catecholamines and serotonin by micellar electrokinetic chromatography with laser-induced fluorescence detection

6-Oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein, a new synthesized fluorescent reagent, was established for the first time as a label for the sensitive analysis of catecholamines (CAs) and serotonin (5-HT) by micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence detection. After careful study on the derivatization conditions such as pH value, reagent concentration, temperature and reaction time, the labeling reaction was accomplished as quickly as 7 min with stable yield. The separation parameters for the CAs and 5-HT were also optimized in detail. The derivatives were baseline separated in a running buffer containing 30 mM boric acid and 15 mM sodium dodeculsulfate at pH 9.0. The detection limits ranged from 5 x 10(-10) to 2 x 10(-9) M (signal-to-noise ratio = 3). The rapid and sensitive method was also applied to the determination of the CAs and 5-HT of urine samples.     

方波激发非接触式电导检测器信噪比的影响因素研究

研制了用于毛细管电泳的方波激发非接触式电导检测器(CCD)并详细考查了对检测器信噪比影响较大的几个重要因素,包括激发频率和运算放大器、基线分压电路和A/D转换器、数字低通滤波;以及常见缓冲体系2-(N-吗啡啉)-乙磺酸/组氨酸(MES/His)体系中电解质浓度,添加剂十六烷基三甲基溴化铵(CTAB)对金属离子的检出信号的影响。所得的结果对于该类检测器的设计和优化具有参考价值。     

Critical aspects of the determination of pentafluorobenzyl derivatives of aldehydes by gas chromatography with electron-capture or mass spectrometric detection: Validation of an optimized strategy for the determination of oxygen-related odor-active aldehydes in wine

This work presents a thorough study of some aspects critical to the quantitative performance of methods for the determination of volatile aldehydes previously derivatized to pentafluorobenzyl hydroxylamine oximes. The conclusions of the study are further applied to the validation of an optimized procedure for the determination of oxidation-related aldehydes from wine. Aspects studied include the chromatographic injection, the analytical performance of electron-capture detection (ECD) or MS detection, and the way in which the derivatization is carried out. Different injection techniques have been optimized and compared (classical splitless-hot injection; cold splitless; and large volume solvent split injection). All of them were carried out in a programmed temperature vaporization (PTV) injector, with a 350 microL internal volume liner (3.4 mm internal diameter). Classical splitless injection of hexane extracts is troublesome and requires large carrier gas flows (>10 mL min(-1)). Cold splitless injection was clearly superior. Large volume solvent split injection has been also optimized. ECD has been found to lack the necessary selectivity for the determination of unsaturated aldehydes in wine, although the quantitation of several minor aldehydes is possible. MS detection has proven to be superior for the determination of these compounds in wine. The way in which the reagent is applied is also critical and for the case of wine is important to ensure that the reagent is applied after some of the major wine aldehydes have been eliminated. The finally proposed method is extremely sensitive. Method detection limits ranged from 0.002 microg L(-1) (for unsaturated aldehydes) to 0.73 microg L(-1) (for phenylacetaldehyde) and precision (measured as relative standard deviation) is < or =6% in all cases. The method makes it possible to determine quantitatively in a single run the wine aldehydes with sensory significance (isobutyraldehyde, 2-methylbutanal, isovaleraldehyde, (E)-2-hexenal, (E)-2-heptenal, (E)-2-octenal, (E)-2-nonenal, methional and phenylacetaldehyde).     

Estimation of239Pu in urine,influence of Sulkowich reagent

Plutonium is known to be co-precipitated with Sulkowich reagent as calcium ammonium oxalate. In adopting this technique for bio-assay of plutonium, its accuracy depends on the self-absorption of the resulting precipitate in each urine sample. Pu recovery experiments were carried out with varying concentration of Ca and Mg, using different volumes of Sulkowich reagent. When the sample volume is 500 ml, Pu in urine can be estimated with an accuracy and precision of 74.38%±7.4%; with a detection limit of 0.06 Bq (1.6 pCi) per dm³. An attempt was made to improve the detection limit, accuracy and precision values. The results obtained are presented in this paper.     

Indirect enantioseparation of alpha-amino acids by reversed-phase liquid chromatography using new chiral derivatizing reagents synthesized from s-triazine chloride

Ten chiral dichloro- and monochloro-s-triazines were prepared by the nucleophilic displacement of chlorine atom(s) in s-triazine chloride and its 6-methoxy derivative with different amino acid amides. Dichloro-s-triazines (DCTs) were used as CDRs for derivatization of alpha-amino acids under basic conditions at room temperature (30 degrees C) while derivatization with monochloro-s-triazine (MCT) reagents was carried out at 80 degrees C. The resultant diastereomers were separated on a reversed-phase C(18) column using mixtures of acetonitrile and aqueous-trifluoroacetic acid (TFA). The separation results for the two were compared. One DCT reagent was optimized for derivatization kinetics with respect to the effects of pH, reagent excess, temperature and reaction time on derivatization yield. In most of the cases, DCT reagents provided better separation of diastereomers in comparison to MCT reagents. One DCT reagent was also validated for limit of detection, linearity, recovery and robustness. Effects of structural modifications in reagents on chromatographic properties were investigated. Separation mechanism of diastereomers was proposed in light of both MCT and DCT reagents.     

Synthesis of succinimidyl-(S)-naproxen ester and its application for indirect enantioresolution of penicillamine by reversed-phase high-performance liquid chromatography

Synthesis of N-succinimidyl-(S)-2-(6-methoxynaphth-2-yl) propionate was carried out by the reaction of (S)-naproxen with N-hydroxysuccinimide in the presence of dicyclohexyl carbodiimide. It was characterized and was used as a chiral derivatizing reagent, under mild conditions, to form diastereomers of dl-penicillamine which were resolved by reversed-phase high-performance liquid chromatography using triethyl ammonium phosphate buffer (pH 4.0, 5mM)-acetonitrile (linear gradient (30min) of acetonitrile from 30 to 70%). Excellent separation was achieved with gradient mobile phase. The detection limit was at pmol level.     

Metal Analyses in Environmental and Pharmaceutical Samples by Capillary Electrophoresis with Methyl 3‐Amino‐3‐(pyridin‐3‐yl)propanoate Dihydrochloride as a New Ion‐Pairing Reagent

Separation and determination of some common metal ions was achieved with methyl 3‐amino‐3‐(pyridin‐3‐yl)propanoate dihydrochloride (MAPP) as an ion‐pairing reagent and pyridine as a detectable counter‐ion for indirect UV detection at 254 nm. The effects of the complexing reagent and chromophore concentrations, applied voltage, and organic solvent content on the separation were investigated. The optimized separation was carried out in a running electrolyte containing 16 mM MAPP and 20 mM pyridine at pH 4.0 and was successfully applied to the qualitative and quantitative analysis of Li⁺, Na⁺, Mg²⁺, Ca²⁺, Ba²⁺, Ni²⁺, and Zn²⁺ in pharmaceutical vitamin preparations and various water samples.     

Fluorimetric sequential injection determination of magnesium using 8-hydroxiquinoline-5-sulfonic acid in a micellar medium

A fluorimetric sequential injection method for the determination of magnesium is proposed. The system is based on the complex formation between Mg(II) and 8-hydroxyquinoline-5-sulfonic acid (HQS). The reaction was carried out in the presence of ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) as a masking agent. Cetyltrimethylammonium chloride (HTAC) was employed as a fluorescence enhancer. The influence of several variables, such as reagent concentration, volumes and pH has been investigated. The reagent was prepared in a 0.1 M Tris-HCl buffer solution (pH 9). A detection limit of 12 mug l(-1) magnesium was obtained. The proposed method was applied to the determination of magnesium in natural waters.     

Liquid Chromatographic Analysis of Tryptophan and Serotonin Metabolites. Comparison of UV,Electrochemical and Spectrofluorimetric Detection

Abstract

Tryptophan and five of its indolic metabolites have been separated by reversed-phase high performance liquid chromatography. The isocratic chromatographic system consisted of a column (20 × 0.46 cm i.d.) packed with RSil C18 HL5 and a methanol/NaClO₄ O.2 M-HClO₄ (pH 1.4) mobile phase.

Three detection methods were tested : UV detection, fluorometric detection and electrochemical detection.

The limits of detection were found to be 4 picomoles (serotonin, 5 HTrp) and 20 pmoles (Trp, 5 HIAA, IAA, N-acetyl Trp) in the case of UV detection; 1 pmole (serotonin), 4 pmoles (5H-Trp) and 10 pmoles (5 HIAA, Trp, IAA, N-acetyl Trp) for fluorometric detection; and 1 pmole for electrochemical detection.

Analysis of human plasma were carried out using the above three detection methods to compare their relative specificity.     

Determination of trace levels of ethylenethiourea by HPLC following derivatisation with phenacyl halides

Summary The HPLC determination of ethylenethiourea (ETU, imidazolidine-2-thione) has been carried out by derivatisation with phenacyl halides to give imidazole-[2,1-b]thiazoles, which were separated on a polystyrenedivinylbenzene (PS-DVB) column. A number of different phenacyl halides including p-phenyl-, and naphthyl, were studied and the p-nitro reagent was selected as the most suitable for residue analysis. The analysis was more sensitive than direct HPLC analysis and had a limit of detection of 0.04 ng ETU. Presented at the 15th International Symposium on Chromatography, Nürnberg, October 1984     

Rapid and sensitive determination of tryptophan, serotonin and psychoactive tryptamines by thin-layer chromatography/fluorescence detection

A rapid, sensitive and selective method for the determination of tryptophan (Trp), serotonin (5-HT) and psychoactive tryptamines (PATs) by thin-layer chromatography (TLC) with fluorescence detection is proposed. These compounds form fluorophores on the developing plate by heating after spraying with sodium hypochlorite, hydrogen peroxide or potassium hexacyanoferrate(III)-sodium hydroxide reagent. Fluorescent spots (vivid blue) were observed by irradiation with ultraviolet light (365 nm). The detection limits of Trp, 5-HT and PATs were in the range from 0.01 microg to 0.06 microg. This method was effectively applied to the detection of confiscated PAT powder and PAT in abusers' urine samples.     

Detection of chlordanes by positive ion chemical ionization in an ion trap: a comparative [correction of comparitive] study of the non-conventional reagents acetonitrile, acrylonitrile and dichloromethane

The detection of some chlordane compounds (heptachlor, cis-/trans-chlordane and cis-/trans-nonachlor) by positive ion chemical ionization (PICI) in an ion trap was studied using acetonitrile, acrylonitrile and dichloromethane as non-conventional reagent gases. These reagent gases initiated specific fragmentation reactions and resulted in different response factors. All reagent gases enabled detection limits in the low-pg range for heptachlor, whereas the detection limits of cis-/trans-chlordane and cis-/trans-nonachlor were in the mid-pg range. Additionally, the acetonitrile and dichloromethane PICI mass spectra of the cis- and trans-stereoisomers of chlordane and nonachlor were different.     

Ion chromatographic determination of trace level phosphorus in purified quartz

Trace levels of phosphorus in purified quartz are determined by ion chromatography. In situ reagent purification, matrix digestion and oxidation of phosphorus to orthophosphate ion are carried out simultaneously in a vapour phase digestion (VPD) assembly using a mixture of HF, HNO3 and H2O2. A drastic reduction (475 times) in phosphate blank from reagents (HF/H2O2) was achieved in the VPD through in situ purification of the reagent. The residues remaining after volatilisation (solvent/matrix), mostly consisting of insoluble phosphate/fluoride salts of divalent and trivalent cations, were solubilised by ion-exchange dissolution. Phosphate was analysed on the IonPac AS17 column with suppressed conductivity detection. The results of the ion chromatography (IC) method were compared with a spectrophotometric method. Accuracy was evaluated by analysing a certified reference material (silicon, NIST 57a). The method detection limit was 0.05 microg g(-1).     

Simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue by high-performance liquid chromatography with electrochemical detection

A simple method for the simultaneous determination of acetylcholine, choline, noradrenaline, dopamine and serotonin in brain tissue was developed by using high-performance liquid chromatography with electrochemical detection. These compounds are analysed in a single chromatographic run within 30 min with a simple sample clean-up procedure. The detection system consists of two electrochemical detector cells aligned in series: a glassy-carbon electrode for catecholamines and serotonin, and a platinum electrode for acetylcholine and choline. For the detection of the latter compounds, they were converted enzymatically into hydrogen peroxide through a column reactor with immobilized acetylcholinesterase and choline oxidase. A column of boronic acid gel was placed just ahead of the immobilized enzyme column to remove catecholamines, which caused interfering responses on the platinum electrode. Two equivalent analytical columns and a column switching were employed to speed up the serotonin assay. Simultaneous determination of these major neurotransmitters in rat brain regions was successfully carried out with the system described.     

微波辅助萃取-衍生气相色谱-电子捕获检测器法测定电子电气产品中的四溴双酚A

以N,O-双(三甲基硅烷基)三氟乙酰胺(BSTFA)作衍生化试剂,采用微波辅助萃取-衍生气相色谱-电子捕获检测器法(GC-ECD)测定电子电气产品中的四溴双酚A。使用V(甲苯)∶V(甲醇)=10∶1微波辅助萃取电子电气样品,用正己烷沉淀萃取液中的高聚物,净化后,将萃取液进行衍生化反应,采用ECD检测器进行定量测定。对衍生化时间、衍生化温度、衍生化试剂用量、沉淀试剂用量等前处理条件进行了优化,并进行了线性、回收率、精密度等试验。结果表明,方法线性范围为0.005~5 mg/L,相关系数为0.9985,方法的检测下限为0.02 mg/kg。采用样品加标的方式进行四溴双酚A的精密度及回收率实验,回收率在81.7%~110%之间,相对标准偏差RSD(n=7)小于7.2%。所建方法能很好的应用于电子电气产品中四溴双酚A的检测。     

HPLC determination of serotonin and its metabolites from human platelet-rich plasma; shift to 5-hydroxytryptophol formation following alcohol consumption

A sensitive, simple, and reliable high-performance liquid chromatographic method with electrochemical detection is developed for the measurement of four natural products, the serotonin-related indols from human platelet-rich plasma (PRP) using N-methylserotonin as internal standard. Separation of serotonin (5HT), 5-hydroxytryptophan (5HTP), 5-hydroxytryptophol (5HTOL), and 5-hydroxyindole-acetic acid (5HIAA) is carried out on Supelcosil LC-18DB stationary phase. A mixture of 48mM citric acid, 28mM sodium phosphate dibasic, 0.027mM Na(2)EDTA, and 3% methanol (pH 3.18) serves as the mobile phase. Measurements are carried out at 25 degrees C at E(ox) = 0.65 V. The calibration curves are linear through the range of 10-200 pg/mL. Method validation is performed according to internationally accepted criteria. Blood is collected from healthy controls and schizophrenic subjects. Significantly higher PRP serotonin is measured in schizophrenics; patients with recent alcohol consumption could be characterized with significantly elevated 5HTOL/5HIAA ratio.     

Free and total amino acid composition in blue-green algae

A simple, accurate and reproducible analytical method is described for the extraction and the simultaneous determination of 18 amino acids in different for geographical origin Spirulina alga samples using phenylisothiocianate as derivatizating agent in natural feed. The best experimental hydrolysis conditions have been studied varying the temperature, the time and the hydrolyzing reagent. The separation and the quantitative analysis of the by-products have been carried out by HPLC analysis and UV detection. An amino acid pattern is compared with that proposed by the Food Agriculture Organization (FAO) for an ideal protein and with those of some traditional feed.     

Injection port silylation of γ-hydroxybutyrate and trans-hydroxycrotonic acid: conditions optimisation and characterisation of the di-tert-butyldimethylsilyl derivatives by GC-MS

Silylation is usually carried out on γ-hydroxybutyrate (GHB) for its analysis by Gas Chromatography/Mass Spectrometry (GCMS) and requires potentially long incubation times before injection during which the derivatisation reagent and derivatives (such as trimethyl-silyl compounds) can hydrolyse. Moreover, alternative internal standards (IS) are often useful depending on sample matrices, extraction/purification procedures, commercial availability and price. This study evaluated the possibility of silylating GHB with an injection port derivatisation procedure using N-methyl-N-[tert-butyldimethyl-silyl]trifluoroacetimide (MTBSTFA) with 1% tert-butyldimethylchlorosilane (TBCS) as the derivatisation reagent, producing di-tert-butyldimethyl-silyl derivatives as a novel means of analyzing GHB. In parallel, trans-hydroxycrotonic acid (t-HCA) was investigated as a potential IS for GHB quantification. Analyses were carried out with a temperature programmable injector and the GHB(t-BDMS)(2) and t-HCA(t-BDMS)(2) derivatives were successfully produced, characterised and derivatisation conditions optimised. t-HCA behaved very similarly to GHB through the derivatisation processes and was used as the IS for the determination of urinary endogenous GHB concentrations in human subjects where the method showed a limit of detection of 0.049 μg mL(-1), a limit of quantification of 0.162 μg mL(-1), and a limit of confirmation of 1.33 μg mL(-1), suitable for toxicological GHB concentration determination.     

Electrophoretic determination of biogenic amines in biological fluids

The capabilities of the electrophoretic separation of biogenic amines (adrenalin, noradrenalin, dopamine, serotonin, metanephrine, and normetanephrine) under conditions of capillary zone electrophoresis and micellar electrokinetic chromatography with the introduction of complex-forming agents (18-crown-6, 4,13-diaza-18-crown-6, and sodium dodecyl sulfate as an ion-pair reagent) and acetonitrile as the constituents of a working electrolyte were demonstrated. A technique for the sampling of biological fluids (urine, blood plasma, and serum) with the use of solid-phase extraction on aluminum oxide and a C18 reversed-phase sorbent was developed. The capabilities of various versions of the preconcentration of biogenic amines were determined, which allowed us to decrease the limits of detection by a factor of hundreds.