Simultaneous determination of 23 amino acids and 7 biogenic amines in fermented food samples by liquid chromatography/quadrupole time-of-flight mass spectrometry

A novel liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-Q-TOFMS) method was developed for the simultaneous determination of 23 amino acids and 7 biogenic amines in food samples. These analytes were pre-column derivatized with dansyl chloride and then separated in an Acquity column (1.7 μm; 2.1 mm × 100 mm). The separation of 31 compounds including an internal standard was achieved within 25 min at a flow rate of 0.2 mL/min. The method linearity for each amino acid and biogenic amine had a relatively wide range with r(2)>0.99. The intra- and inter-day precision, expressed as relative standard deviation (RSD), ranged from 1.1 to 4.6% and from 2.0 to 11.2%, respectively. The limit of detection was between 0.005 and 0.4 μg/mL. With a simple dilution, recoveries of around 80-120% were obtained for most of the compounds. No significant matrix effect was observed, and the developed method was successfully applied to the analysis of amino acids and biogenic amines in beer, cheese and sausage samples.     

Development of microwave-assisted derivatization followed by gas chromatography/mass spectrometry for fast determination of amino acids in neonatal blood samples

Analysis of amino acids in blood samples is an important tool for the diagnosis of neonatal amino acid metabolism disorders. In the work, a novel, rapid and sensitive method was developed for the determination of amino acids in neonatal blood samples, which was based on microwave-assisted silylation followed by gas chromatography/mass spectrometry (GC/MS). The amino acids were derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) under microwave irradiation. The controlled reaction was carried out employing BSTFA under conventional heating at 120 degrees C for 30 min. Experimental results show that microwave irradiation can accelerate the derivatization reaction of amino acids with BSFTA, and much shorten analysis time. The method validations (linear range, detection limit, precision and recovery) were studied. Finally, the method was tested by determination of amino acids in neonatal blood by the measurement of their trimethylsilyl derivatives by GC/MS in electron impact (EI) mode. Two biomarkers of L-phenylalanine and L-tyrosine in phenylketonuria (PKU)-positive blood and control blood were quantitatively analyzed by the proposed method. The results demonstrated that microwave-assisted silylation followed by GC/MS is a rapid, simple and sensitive method for amino acid analysis and is also a potential tool for fast screening of neonatal aminoacidurias.     

溶剂转移-气相色谱-质谱法和选择洗脱-气相色谱法测定大蒜中289种农药多残留

以溶剂转移净化为核心步骤,建立了一种适用于大蒜样品中农药多残留分析的前处理方法(方法I),配以一个辅助方法(方法II),构成大蒜中常见289种农药多残留的分析体系(方法I283种,方法II6种)。方法I中,样品用乙腈-水溶液提取,盐析分配,溶剂转移和固相萃取(SPE)净化后进行气相色谱-质谱(GC-MS)分析;方法II中,样品用无水Na2SO4配合乙酸乙酯均质研磨,超声波辅助提取,提取液经Primary Secondary Amine (PSA)粉末分散固相萃取和LC-Si柱选择洗脱净化后进行GC分析。GC-MS采用选择离子监测(SIM)方式,GC采用火焰光度检测器(FPD)检测,外标法定量。方法简便、快速,通过优化前处理和上机条件,在最优条件下进行测试,方法的定量限(S/N≥10)为0.01～0.05 mg/kg。方法I中,在加标水平为0.02、0.20 mg/kg时,回收率为52%～163%,其中回收率在70%～120%之间的占88%,相对标准偏差为2.4%～18%;方法II中,在加标水平为0.01、0.02、0.10、0.20 mg/kg时,回收率为70%～111%,相对标准偏差为3.2%～9.3%。详细描述了实验模型的构建,并对GC-MS灵敏度的提高提出了新的见解。该方法准确、灵敏、快速,可满足大蒜中多种农药残留的检测要求。     

Rapid determination of amino acids in foods by hydrophilic interaction liquid chromatography coupled to high-resolution mass spectrometry

This study describes a rapid and sensitive analytical method for the determination of amino acids in foods and drinks. The method entailed dilution or extraction of amino acids from foods using the mixture of acetonitrile and 0.1% aqueous formic acid (50:50, v/v). Chromatographic separation of underivatized amino acids was performed using a hydrophilic interaction liquid chromatography within a runtime of 6 min. Both hydrophobicity and charge of the side chain played important roles on the elution order of amino acids under the chromatographic conditions. High-resolution mass spectrometry allowed qualitative and quantitative detection of amino acids in complex food matrices. Its response was found linear over a concentration range of 0.25-10 μg/ml. The method could be successfully applied to various foods and drinks to profile individual amino acids. Mean percentage recoveries of amino acids from different matrices were 88.5% or higher with residual standard deviation of less than 5.0%.     

Rapid determination and confirmation of biogenic amines in tuna loin by gas chromatography/mass spectrometry using ethylchloroformate derivative

A gas chromatography-mass spectrometry (GC/MS) method is described for the easy rapid determination and simultaneous confirmation of the biogenic amines putrescine (PUT), cadaverine (CAD), histamine (HTA), and spermidine (SPD) in fresh frozen tuna loin. The method can also be used to monitor tyramine (TYR). The method involves homogenization of fish tissue, extraction of biogenic amines into trichloroacetic acid solution, centrifugation, alkalization, and derivatization of supernatant with ethylchloroformate. All seafood species were fortified to contain 2.5, 5.0, 10.0, 12.5, and 25.0 microg/g (ppm) PUT, CAD, and SPD; and 10.0, 20.0, 40.0, 50.0, and 100.0 microg/g (ppm) HTA. Determination was based on standard curves for each analyte using peak areas with matrix standards equivalent to a concentration range bracketing the spike level. A set of 5 matrix controls (unfortified tuna tissue) was also analyzed; only endogenous SPD was found in all samples. The interassay average recoveries ranged from 57 to 79% across analytes and spike levels.     

Simultaneous profiling analysis of alkylphenols and amines by gas chromatography and gas chromatography–mass spectrometry

Two-phase O-ethoxycarbonylation was performed to alkylphenols in acidic solution with ethyl chloroformate present in dichloromethane phase fortified with triethylamine with subsequent N-ethoxycarbonylation of amines after adjusting to alkaline pH. The resulting ethoxycarbonyl derivatives were subjected to pentafluoropropionylation, clean-up and concentration for analysis by gas chromatography and gas chromatography–mass spectrometry. The present method was linear (r ≥ 0.9959) in the range of 0.5–10.0 μg ml^−l with good precision (≤9.5%) and accuracy (−8.9 to 9.5%) for 20 phenols and 27 amines examined, allowing simultaneous screening for a total of one alkylphenol and four amines from wine and beer.     

Determination of novel aromatic amines in environmental samples by gas chromatography/mass spectrometry

A number of compounds belonging to five chemical classes of aromatic amines were identified by electron impact mass spectrometry, both in the vapor phase and in suspended particulate matter collected in an area subjected to heavy traffic. This paper presents the mass spectra of new aromatic amines, proposed mechanisms for formation of the major ions obtained by electron impact, concentrations of each class in 13 environmental samples, and the distribution of the individual compounds of the major chemical classes (greater than 80% by weight).     

Automated capillary liquid chromatography for simultaneous determination of neuroactive amines and amino acids

A method for the separation and quantitative determination of neuroactive amino acids (aspartate, glutamate, citrulline, arginine, glycine, taurine, gamma-aminobutyric acid) and neuroactive amines (noradrenaline, dopamine and serotonin) in a single chromatographic analysis is presented. The method is based on pre-column derivatization with o-phthalaldehyde and tert.-butyl thiol, on-column preconcentration and separation using 50 microns I.D. packed capillary columns, and detection by amperometry. Mass limits of detection are 80-900 amol for all neurotransmitters with RSDs of 0.71 and 4.6% or better for retention time and peak area, respectively. The method was demonstrated by application to the determination of neurotransmitters in microdialysis samples collected from striatum of live rats and tissue samples extracted from butterfly brains.     

Characterization of sulfonated azo dyes and aromatic amines by pyrolysis gas chromatography/mass spectrometry

Sixteen sulfonated and unsulfonated azo dyes as well as eleven sulfonated and unsulfonated aromatic amines were analyzed and qualitatively characterized by means of pyrolysis gas chromatography/mass spectrometry at different temperatures. Aniline and aminonaphthalene were found to be the dominant pyrolysis products of sulfonated aromatic amines and dyes. Azo dye and dye class specific key compounds such as benzidine, vinyl-p-base and 4-aminoazobenzene could be identified by pyrolysis gas chromatography/mass spectrometry of commercial acid, cationic, direct, reactive and solvent dyes. 500 degrees C was the optimal pyrolysis temperature for most of the pyrolyzed compounds. The method was applied to a dried sample of a textile wastewater concentrate from a dyeing process. Reactive azo dyes of the group of Remazol dyes and anthraquinone dyes could be identified as the major compounds of the sample. The finding of caprolactam (a printing additive) suggests that the wastewater contained effluent from a process of heat-activated printing with reactive dyes. p-Chloraniline, a banned aromatic amine, was identified. Chemical reduction of the wastewater sample prior to pyrolysis resulted in the release of volatile aromatic amines and aided the classification of several products of pyrolysis.     

Overview of methods for the determination of trans fatty acids by gas chromatography, silver-ion thin-layer chromatography, silver-ion liquid chromatography, and gas chromatography/mass spectrometry

trans Isomers of naturally occurring cis-unsaturated fatty acids are produced when liquid vegetable oils or marine oils are partially hydrogenated to produce margarine, shortenings, and other hardened-fat products. Isomeric trans fatty acids are also formed in the intestinal tract of ruminants, and they appear in small amounts in dairy products and ruminant meat. Currently, satisfactory analyses for the fatty acid profiles of fats containing trans fatty acids are obtained by gas chromatography (GC) using capillary columns coated with highly polar cyanosilicone stationary phases. In capillary GC methods, the key limitation has been the incomplete separation of trans-monoenoic acid isomers from their cis isomers; however, recent reports have demonstrated that improvements in separation are attainable with the use of 100 m columns. In these columns, there is very little overlap of cis and trans isomers. More accurate trans fatty acid analyses can be obtained by coupling GC with either silver-nitrate thin-layer chromatography or silver-nitrate liquid chromatography.     

Triazolopyridine,a new derivative for the structural determination of fatty acids by gas chromatography/mass spectrometry

Triazolopyridine is proposed as a new derivative for the location of branches and double bonds in fatty acids. It is compared with previously used derivatives and found to be clearly superior to the pyrrolidide and slightly superior to the β-picolinyl ester with regard to the structure-specific fragmentation pattern. It is prepared by reacting the activated acid with 2-hydrazinopyridine followed by cyclization.     

Rapid determination of amino acids in neonatal blood samples based on derivatization with isobutyl chloroformate followed by solid-phase microextraction and gas chromatography/mass spectrometry

The purpose of this study was to develop a simple, rapid and sensitive analytical method for determination of amino acids in neonatal blood samples. The developed method involves the employment of derivatization and a solid-phase microextraction (SPME) technique together with gas chromatography/mass spectrometry (GC/MS). Amino acids in blood samples were derivatized by a mixture of isobutyl chloroformate, methanol and pyridine, and the N(O,S)-alkoxycarbonyl alkyl esters thus formed were headspace extracted by a SPME fiber. Finally, the extracted analytes on the fiber were desorbed and detected by GC/MS in electron impact (EI) mode. L-Valine, L-leucine, L-isoleucine, L-phenylanaline and L-tyrosine in blood samples were quantitatively analyzed by measurement of the corresponding N(O,S)-alkoxycarbonyl alkyl esters using an external standard method. SPME conditions were optimized, and the method was validated. The method was applied to diagnosis of neonatal phenylkenuria (PKU) and maple syrup urine disease (MSUD) by the analyses of five amino acids in blood samples. The results showed that the proposed method is a potentially powerful tool for simultaneous screening for neonatal PKU and MSUD.     

Separation and determination of seleno amino acids using gas chromatography hyphenated with inductively coupled plasma mass spectrometry after hollow fiber liquid phase microextraction

A new derivatization–extraction method for preconcentration of seleno amino acids using hollow fiber liquid phase microextraction (HF‐LPME) was developed for the separation and determination of seleno amino acids in biological samples by gas chromatography–inductively coupled plasma mass spectrometry (GC–ICP‐MS). Derivatization was performed with ethyl chloroformate (ECF) to improve the volatility of seleno amino acids. Parameters influencing microextraction, including extraction solvent, pH of sample solution, extraction time, stirring speed, and inorganic salt concentration have been investigated. Under the optimal conditions, the limits of detection (LODs) obtained for Se‐methyl‐selenocysteine (SeMeCys), selenomethionine (SeMet), and selenoethionine (SeEth) were 23, 15, and 11 ng Se l⁻¹, respectively. The relative standard deviations (RSDs) were 14.6%, 16.4%, and 19.4% for SeMeCys, SeMet, and SeEth (c = 1.0 ng ml⁻¹, n = 7), respectively, and the RSDs for SeMeCys, SeMet could be improved obviously if SeEth was utilized as the internal standard. The proposed method was applied for the determination of seleno amino acids in extracts of garlic, cabbage, and mushroom samples, and the recoveries for the spiked samples were in the range of 96.8–108% and 93.4–115% with and without the use of SeEth as internal standard. The developed method was also applied to the analysis of SeMet in a certified reference material of SELM‐1 yeast and the determined value is in good agreement with the certified value. Copyright © 2008 John Wiley & Sons, Ltd.     

Direct determination of enantiomeric purity of FMOC amino acids with high performance liquid chromatography

Summary Analytical methods are described which allow a direct determination of enantiomeric purity of seventeen FMOC amino acids commonly used in peptide synthesis. The corresponding ester derivatives can be separated directly on cellulose tris(3,5-dimethylphenylcarbamate) (Chiralcel-OD). The methods are suitable for primary as well as secondary FMOC amino acids. The presence of a highly sensitive fluorescence moiety within the molecule, in combination with large separation factors (-values between 1.5–2.2) allowed for a general detection limit below 0.05%. In several cases the antipode has been determined in the ppm-range. An interesting result has been observed with respect to the elution order of the FMOC amino acid esters. The elution order of the Trp enantiomers is opposite to that obtained with the other amino acids. This is contrary to the generally held belief that elution order is identical within a homologous series of racemates when chromatographed under identical conditions on the same chiral stationary phase. In addition, the inversion of elution of the Pro enantiomers depending on the estertype indicates a competition of different separation mechanisms.     

Determination of plasma amino acids by gas chromatography

A complete methodology, incorporating a novel clean-up technique, for quantitative determination of amino acids in plasma by gas chromatography is described. Glucose, a component causing major analytical interference, is removed by an enzymic reaction included in the pre-chromatographic clean-up. The procedure for derivatisation of amino acid standards is shown to be reproducible down to a level of 2.5 micrograms for each amino acid, relative standard deviations for all amino acids except arginine and histidine being 3% or lower. For the entire procedure applied to plasma, relative standard deviations for most amino acids are below 5% with recoveries ranging from 90 to 120%. Normal values, obtained using eighteen plasma samples, are in reasonable agreement with published data. Plasma amino acid values were determined simultaneously by gas chromatographic and ion-exchange chromatographic techniques. Statistical evaluation shows there to be no significant difference between corresponding values for eleven amino acids. Values for tyrosine, histidine and particularly phenylalanine show significant differences (p less than 0.001).