Identification and Determination of Tele-Methylhistamine in Cerebrospinal Fluid by Gas Chromatography-Mass Spectrometry

Abstract: The presence of tele-methylhistamine in human cerebrospinal fluid has been established. The concentration was determined with the use of deuterated tele-methylhistamine. The preparation of the deuterated standard is described. The concentration range in samples from neuropsychiatric patients was 0.1-2.5 ng/ml. The structure of the pentafluoropropionyl derivative used for gas chromatography was studied with the aid of proton nuclear magnetic resonance spectroscopy.     

Determination of 1-Methylimidazole-4-Acetic Acid in Brain Tissue from Rat and Mouse

The concentration of the histamine metabolite 1-methylimidazole-4-acetic acid was determined in brain tissue from rat and mouse with a gas chromatographic-mass spectrometric method. Mouse brain contained 1.7-3.2 nmol/g, depending on the strain. The concentration in cerebrum from Sprague-Dawley rats was 1.2 nmol/g, whereas cerebellum contained 0.24 nmol/g. The concentration of tele-methylhistamine in mouse brain was 1.4-2.2 nmol/g. The concentration of 1-methylimidazole-4-acetic acid in rat brain after death did not change significantly during 2 h at room temperature.     

Determination of N-Acetylaspartic Acid in Human Cerebrospinal Fluid by Gas Chromatography–Mass Spectrometry

N-Acetyl-L-aspartic acid was identified and determined in human cerebrospinal fluid. The concentration in lumbar fluid was about 2 nmol/ml and about 20 nmol/ml in ventricular fluid. There was no difference between healthy subjects and schizophrenic patients.     

Gas Chromatographic-Mass Spectrometric Determination of myo-Inositol in Human Cerebrospinal Fluid

The concentration of free myo-inositol in CSF was determined with a gas chromatographic-mass spectrometric method using deuterated myo-inositol as an internal standard after conversion to the hexa-O-acetyl derivative with acetic anhydride and pyridine. Twenty microliters of CSF is sufficient for the analysis which has a coefficient of variation of 9%. Identical analytical results were obtained on two different mass numbers. Schizophrenic patients were compared with healthy control persons. In addition, patients with rheumatoid arthritis or with neurological illnesses were studied. No consistent differences related to the illness could be found. The mean concentration of myo-inositol was about 25 micrograms/ml. Treatment of schizophrenic patients with chlorpromazine or sulpiride had no significant effect on the concentration of myo-inositol in CSF.     

Presence and Measurement of Imidazoleacetic Acid, a γ-Aminobutyric Acid Agonist, in Rat Brain and Human Cerebrospinal Fluid

Imidazoleacetic acid (IAA) was unequivocally demonstrated in rat brain, human CSF, and human plasma by a gas chromatographic-mass spectrometric method that can reliably quantify as little as 8 pmol, i.e., 1 ng. Owing to tautomerism of the imidazole ring, IAA and [15N, 15N]IAA, the internal standard, each formed two chromatographically distinct isomers after derivatization of the ring nitrogens with either ethyl chloroformate or methyl chloroformate. The isomers of n-butyl(N-ethoxycarbonyl)imidazole acetate and n-butyl(N-methoxycarbonyl)imidazole acetate were identified by analysis with methane chemical ionization and electron impact ionization of molecular and fragment ions. The levels (mean +/- SEM) of free IAA were 140 +/- 14 pmol/g and 2.7 +/- 0.2 pmol/ml in brains of untreated rats and human lumbar CSF, respectively. Mean levels of IAA in brains of anesthetized rats, perfused free of blood, did not differ significantly from mean levels of anesthetized, nonperfused controls or from untreated rats. The source or sources of IAA in brain and CSF are unknown. Because IAA is a potent agonist at gamma-aminobutyrate receptors, it merits examination as a regulator in brain.     

Demonstration of Conjugated Dopamine in Monkey CSF by Gas Chromatography-Mass Spectrometry

Abstract: A method for measuring unconjugated and conjugated dopamine in body tissues and fluids is described. Conjugated dopamine was hydrolyzed in acid to unconjugated dopamine, separated from the sample matrix by alumina chromatography, and assayed by gas chromatography-mass spectrometry. Conjugated dopamine was detected in greater concentrations than unconjugated dopamine in CSF taken from lateral ventricle or thecal sac of the Rhesus monkey. Haloperidol administration did not increase the levels of conjugated dopamine in lumbar CSF.     

缬草油化学成份GC/MS分析研究

利用GC、GC/MS对缬草油的化学成分进行分析 ,共鉴定出 6 5种化合物 ,其中有 2 6种物质为相关文献中首次报道。所介绍的分析方法可用于生产中的质量监控和常规分析 ,分析结果可为配方、产品开发和调香等提供指导     

The Significance of Homovanillic Acid and 3,4-Dihydroxyphenylacetic Acid Concentrations in Human Lumbar Cerebrospinal Fluid

The concentrations of the acidic dopamine (DA) catabolites homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) measured in human CSF are supposed to reflect the "turnover" of DA in the brain. The notion of "turnover" is, however, not synonymous with impulse nerve activity in the dopaminergic systems. Significant amounts of DOPAC and HVA could, indeed, be demonstrated in brain structures wherein dopaminergic innervation has not been documented. It must also be noted that DA is not only a neurotransmitter itself, but also a precursor of norepinephrine and epinephrine. Furthermore, in lumbar CSF, levels of biogenic amine catabolites partially reflect metabolism in the spinal cord and may have limited relevance to neurotransmission in the brain. To elucidate these points further, we determined the concentrations of DOPAC and HVA in 22 areas of six human brains and eight levels of six human spinal cords. The data were correlated with the concentration of DA. Quantitative determinations were done using HPLC with electrochemical detection, after solvent and ion-pair extraction. In this study, significant amounts of both DOPAC and HVA were demonstrated in brain structures not previously associated with dopaminergic innervation. The relatively lower DA concentration in these structures suggests that in these regions, the DOPAC and HVA concentrations are unrelated to dopaminergic neurotransmission. The possible role of capillary walls and glial cells in the catabolism of DA must be further evaluated. The demonstration of DOPAC and HVA in the spinal cord is another argument against the hypothesis that CSF levels of HVA and DOPAC reflect closely the activity of the dopaminergic systems in the brain.     

气相色谱质谱法测定食品包装材料中24种邻苯二甲酸酯(英文)

目的:建立同时检测食品包装材料中24种邻苯二甲酸酯类化合物的气相色谱质谱法(GC-MS)分析方法。方法:用正己烷提取包装材料,GC-MS选择离子监测模式(SIM)测定,运用气质联用仪测定24种邻苯二甲酸酯类物质。结果:24种邻苯二甲酸酯类物质的线性范围为0.05 mg/L~10 mg/L,除了邻苯二甲酸二异壬酯(DINP)和邻苯二甲酸二异癸酯(DIDP)为0.5 mg/L~10 mg/L,相关系数(r2)除DINP、DIDP外均大于0.99,方法的检出限(信噪比为3)为0.002 mg/kg~0.05 mg/kg,在食品包装材料基质中3个加标水平的平均回收率为85.2%~108%,相对标准偏差(RSD,n=6)为5.9%~10.2%。结论:该方法快速、灵敏、准确可靠,适用于食品包装材料中邻苯二甲酸酯类化合物的分析检测。     

Effect of Freezing on γ-Aminobutyric Acid Levels in Human Cerebrospinal Fluid

Abstract Using a radioreceptor assay, the concentration of γ -aminobutyric acid (GABA) in human cerebrospinal fluid (CSF) was found to be elevated significantly following a single deep-freeze to –70°C and thaw. Mean CSF GABA (± SD) in unfrozen CSF was 173 ± 73 pmol/ml ( n = 24). After a single deep-freeze, the mean level was 243 ± 106 pmol/ml ( p < 0.02). Subsequent freeze-thaw cycles resulted in further irregular and unpredictable elevations in CSF GABA. Mean level after two freezes was 379 ± 125 pmol/ml and after three freezes 654 ± 411 pmol/ml. These changes could result in the incorrect interpretation of results in patients suffering from neurological diseases.     

Distribution of Putrescine in Rat Brain Measured by Gas Chromatography-Mass Spectrometry

We measured putrescine levels in minute sites of single rat brains using a sensitive, specific assay involving gas chromatography-mass spectrometry. The putrescine level was measured in 20 sites of single rat brains: three sites in the cerebral cortex, six sites in the hypothalamus, three sites in the basal ganglia, three sites in the thalamus, three sites in the limbic system, and two sites in the cerebellum. The level of putrescine was very high in the hypothalamus, high in the basal ganglia and limbic system, and low in the thalamus, cerebellum, and two of the three sites in the cerebral cortex. The highest levels were in the anterior hypothalamic area and the lateral hypothalamic area, and the lowest levels were in the vermis and the lobe of the cerebellum.     

Mass Spectrometric Identification of 13C‐Labeled Metabolites During Anaerobic Propanoic Acid Oxidation

Biowaste digestion is a possibility to gain biogas as a renewable fuel source. However, the anaerobic food chain may be disrupted by, e.g., substrate overload or by inhibitors, leading to the accumulation of volatile fatty acids (VFAs), predominantly of propanoic acid (PA). VFA Accumulation may cause a rapid pH decrease, less biogas production, or even a total inhibition. To maintain high biogas productivity or to prevent a collapse of methanogenesis, metabolic properties of the degrading microorganisms must be elucidated, e.g., by investigation of the established pathways for degradation of VFAs. A Dani 3950 headspace system (HS), a Varian 431 gas chromatograph (GC), and a Varian 210 mass spectrometer (MS) have been combined to quantify and specifically identify metabolites of PA oxidation. The use of [1‐¹³C]‐labeled PA as a carbon source for microorganisms allows differentiation between the methyl‐malonyl‐CoA or the C(6)‐dismutation pathway, both resulting in AcOH production. Appearance of the ¹³C‐moiety either in the COO or Me group of AcO can easily be detected by MS. The methyl‐malonyl‐CoA pathway was successfully identified as the only pathway of PA degradation by organisms in a lab‐scale anaerobic digester. A similar approach can be applied to any degradation pathway involving VFAs.     

Metabolism of 15NH3 in Organotypic Cerebellar Explants and Cultured Astrocytes: Studies with Gas Chromatography-Mass Spectrometry

Gas chromatography-mass spectrometry was used to study the metabolism of 15NH3 in organotypic cerebellar explants and cultured astrocyte monolayers. A steady-state level of 15NH3 was present by 1 min in both systems. Steady-state labeling in L-[amide-15N] glutamine, L-[15N]alanine, L-[15N]glutamate, and L-[15N]aspartate was attained by 1 min after 15NH3 addition in the organotypic cerebellar explants and by approximately 5 min in the cultured astrocytes. No measurable 15N labeling was noted in either glycine or serine in either system.     

GC/MS分析血浆中丁丙诺啡

目的:建立血浆中丁丙诺啡GC/MS分析方法。方法:血浆中丁丙诺啡,加入内标长春西汀,加pH 7缓冲溶液,用三氯甲烷提取,提取物经BSTFA衍生化后进行GC/MS分析。结果:方法的线性范围为2～100 g·L~(-1),检出限为1g·L~(-1)。结论:该方法灵敏度高,可用于涉毒案件血浆中丁丙诺啡的分析。     

Assay of 2-Hydroxyputrescine in Various Regions of Rat Brain by Gas Chromatography-Mass Spectrometry

2-Hydroxyputrescine in seven regions of single rat brains was measured with a sensitive, specific assay by gas chromatography-mass spectrometry. The regions were the cerebral cortex, cerebellum, medulla oblongata, hypothalamus, striatum, hippocampus, and midbrain. The level of 2-hydroxyputrescine was very high in the cerebral cortex and cerebellum, high in the medulla oblongata, hypothalamus, and hippocampus, and low in the striatum and midbrain. The level of 2-hydroxyputrescine in the cerebellum was significantly higher than in the striatum and midbrain.     

Increased Superoxide Dismutase Activity in Aged Human Cerebrospinal Fluid and Rat Brain Determined by Electron Spin Resonance Spectrometry Using the Spin Trap Method

Superoxide dismutase (SOD) activity in CSF of patients was determined by electron spin resonance spectrometry using the spin trap method. Variation in SOD activity was found among patients. SOD activity in CSF of subjects increased with age and this was identified as Cu,Zn-SOD activity by electrophoresis. In addition, animal experiments showed that SOD activities were higher in mitochondrial and cytosol fractions of aged rats than in those of adult rats. This finding on aged rat brain validates the increase of SOD activity in aged human CSF.     

Nucleic Acids in Protein Samples Interfere with Phosphopeptide Identification by Immobilized-Metal-Ion Affinity Chromatography and Mass Spectrometry

Immobilized-metal-ion affinity chromatography (IMAC) is used extensively for phosphopeptide enrichment in phosphoproteomics. However, the effect of nucleic acids in protein samples on phosphopeptide enrichment by IMAC has not yet been well clarified. In this study, we demonstrate that IMAC beads possess a strong adsorption of nucleic acids, especially single-stranded or single-stranded-region-containing nucleic acids, leading to approximately 50% loss of phosphopeptides during the process of IMAC enrichment. Therefore, nucleic acids must be removed from protein samples prior to IMAC. Acetonitrile (ACN) precipitation, a simple and efficient procedure, was established to remove nucleic acids from the protein samples. We showed that ACN precipitation approximately doubled the phosphopeptide number identified by IMAC and mass spectrometry, indicating that nucleic acid removal significantly improves the identification of phosphopeptides. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

2-Pyrrolidinone in Human Cerebrospinal Fluid: A Major Constituent of Total γ-Aminobutyric Acid

2-Pyrrolidinone, the lactam of gamma-aminobutyric acid (GABA), is identified as the major constituent of total GABA in human CSF. Structural elucidation was done by mass spectrometry. In lumbar CSF of four patients, 2-pyrrolidinone represented about 54% of GABA found after acid hydrolysis, thus accounting for essentially all of the hitherto unknown GABA fraction in CSF.     

N-Acetyl Aspartic Acid (NAA) and N-Acetyl Aspartylglutamic Acid (NAAG) in Human Ventricular,Subarachnoid, and Lumbar Cerebrospinal Fluid

N-Acetylaspartic and N-acetylaspartylglutamic acid concentrations in human ventricular, subarachnoid and lumbar cerebrospinal fluid were measured by combined gas chromatography-mass spectrometry using selected ion monitoring with deuterated internal standards. N-Acetylaspartate concentrations were in the range 55, 9, and 1 M, respectively; N-acetylaspartylglutamate concentrations in the same fluids were in the range 8, 3 and 4 M, respectively. There did not appear to be any difference in lumbar fluid concentrations of either compound between control subjects, schizophrenic patients, Alzheimer's disease patients and a pooled group of patients with neurological degeneration. Ventricular concentrations of both compounds were greatly increased in deceased patients suggesting that maintenance of their intracellular concentrations is probably energy dependent. The concentrations of these compounds in lumbar cerebrospinal fluid from living, and ventricular cerebrospinal fluid from deceased subjects were weakly correlated with one another. In lumbar fluid neither compound appeared to be correlated with age. Analysis of serially collected lumbar samples from two subjects showed a weak concentration gradient for both compounds. Neither antipsychotic medication nor the acid transport inhibitor probenecid had any effect on lumbar concentrations of either compound. Attempts to use anion exchange high pressure liquid chromatography with UV detection for measurement of the low concentrations of N-acetylaspartate found in cerebrospinal fluid from living subjects were unsuccessful.     

Kinetics of Homovanillie Acid and Determination of Its Production Rate in Humans

Abstract: Homovanillie acid (HVA) labeled with either two or five deuterium atoms (d₂-HVA or d₅-HVA) was used to label the peripheral body pool of endogenous HVA (d₀-HVA) in control humans and in neurological patients. Either d₂- or d₅-HVA was rapidly injected intravenously and concentrations of d₂- or d₅- and d₀-HVA in sequential samples of blood plasma were determined by gas chromatography-mass spectrometry (GC-MS). Parameters describing the distribution and elimination of HVA, as well as its pool size, turnover, and synthesis rate were then calculated. Results indicate that the decline of plasma d₂- or d₅-HVA with time was multiexponential in five out of six subjects. Basal levels of endogenous HVA averaged 14.4 ± 1.3 ng/ml in the control patients and 8.69 ± 2.4 ng/ml in the neurological patients. The biological half-life averaged 71.3 ± 8.0 min in the control subjects and 91.3 ± 15 min in the neurological patients. The apparent volume of distribution of HVA in the body was 31–100 liters. Plasma clearance was 243–679 ml/min. The size of the peripheral body pool, calculated from plasma kinetic parameters, was 392–673 μg. The HVA rate of production, calculated for the whole body, was 166–323 μg/h. This technique can be used to determine accurately the rate of HVA production by the whole body over short time intervals. Since sample size was very limited ( n = 3 per group) and effects of variables such as age and sex on these data have not been excluded, more thorough investigations are needed to document any differences between normal controls and neurological patients.