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11.
A review of sample preparation methods for organic acids in biological fluids, in particular serum and urine, is presented. It covers techniques on organic acid determination without sample preparation, release of organic acids from binding locations, removal of proteins by protein precipitation and ultrafiltration, isolation of the organic acids by liquid-liquid and liquid-solid extraction, purification of the extract, derivatization and pre-fractionation. The various alternative sample preparation steps are compared and critically discussed. Examples of applications including profile analysis of organic acids by gas chromatography (GC), determination of particular organic acids by GC or liquid chromatography and determination of fatty acids as a distinct chemical class of acids demonstrate that the kind of sample preparation chosen depends strongly on the analytical aims. 相似文献
12.
A multipurpose sampler (Gerstel MPS), designed for liquid large volume, gaseous and headspace samples was used for the GC-MS analysis of organic volatiles in human urine. Headspace sampling with a volume-, temperature- and speed-controlled gas-tight syringe was combined with a temperature-controlled cold injection system (CIS) for cold trapping, enrichment and focusing of analytes. Regular 2-ml GC vials filled with 1 ml acidified urine were used as headspace sampling vials. A 100-vial autosampler tray was equipped with an additional temperature and heating time controlled "preheating station" for five vials. Profiles of organic volatiles in human urine were determined and 34 components identified. Trimethylamine (TMA) and 4-heptanone as two metabolites of medical interest were quantified. Calibration curves and intra assay imprecision for 4-heptanone concentrations in the range of 40 to 800 ng/ml showed a correlation coefficient of r = 0.9980 and a relative standard deviation (RSD) between 3.0 and 3.4%. Calibration curves and intra-assay imprecision for TMA concentrations in the range of medical interest from 0.5 to 20 micrograms/ml showed a correlation coefficient of r = 0.9968 and a RSD between 4.1 and 6.8%. The high practicability of the multipurpose sampler for both gaseous and liquid samples together with the here shown good reproducibility and sensitivity make this single CIS-GC-MS system very attractive for routine clinical use in metabolic profiling of organic volatiles (headspace) and non-volatiles (liquid). 相似文献
13.
Zhu L Ostash B Rix U Nur-E-Alam M Mayers A Luzhetskyy A Mendez C Salas JA Bechthold A Fedorenko V Rohr J 《The Journal of organic chemistry》2005,70(2):631-638
The angucycline antibiotic family of the landomycins displays potent antitumor activity. To elucidate early post polyketide synthase (PKS) tailoring steps of the landomycin E biosynthetic pathway in Streptomyces globisporus 1912, the mutant S. globisporus M12 was prepared through gene replacement experiment of lndM2. It encodes an enzyme with putative oxygenase and reductase domains, according to sequencing of the gene and its counterpart lanM2 from S. cyanogenus S136 landomycin A biosynthetic gene cluster. The isolation of the novel shunt products 11-hydroxytetrangomycin and 4-hydroxytetrangomycin along with the well-known angucyclines tetrangomycin and tetrangulol from the culture of S. globisporus M12 provides evidence for the involvement of lndM2 in the early biosynthetic pathway of the landomycins, in particular in the formation of the alicyclic 6-hydroxy function of the landomycin aglycon. We therefore propose LndM2 to be responsible for both hydroxylation of the 6-position and its subsequent reduction. These reactions are necessary before the glycosylation reactions can occur. The results are in agreement with the originally published structure of landomycin but do not support the recently suggested revised structure. 相似文献
14.
H. M. Liebich G. Xu C. Di Stefano R. Lehmann H. U. Häring P. Lu Y. Zhang 《Chromatographia》1997,45(1):396-401
Summary Modified nucleosides excreted in urine have been studied as potential diagnostic markers for cancer and AIDS, and as indicators
for the whole-body turnover of RNA. Until now, reversed-phase (RP) HPLC and, to some extent, immunoassays are the preferred
analytical methods for urinary nucleosides. A new capillary electrophoretic method for the analysis of normal and modified
nucleosides in urine has been developed and optimized in our laboratory. The separation of nucleosides extracted from normal
human urine on phenyl boronic acid affinity chromatography columns was performed in uncoated 565 mm (500 mm to detection window)
× 50 μm i.d. capillary tubing using a 300 mM SDS—25 mM borate—50 mM phosphate buffer (pH 6.7), a 45-s load, a voltage of 7.5
kV (41 μA) and UV detection at 260 and 210 nm. The average recovery of the nucleosides was 91 %. The calibration curves were
linear over all physiological and pathophysiological concentration ranges and the limits of detection were at micromolar levels.
Reproducibility of migration times were better than 1 % (coefficient of variation,CV), and the reproducibilities of the determined concentrations were better than 5 % for standards and 6–15 % for extracted
urine. The developed method was used to quantify 15 normal and modified nucleosides in 25 normal urines to establish reference
ranges. The analysis time was less than 45 min.
Dedicated to Professor E. Bayer on the occasion of his 70th birthday.
Presented at the 21st ISC held in Stuttgart, Germany, 15th–20th September, 1996. 相似文献
15.
Principal component analysis and cluster analysis have been used in AES investigations of fibre-matrix interactions in alumina fibre reinforced MgLi-alloys prepared by high-pressure infiltration. The samples have been fractured in ultra high vacuum to expose surfaces and interfaces without contamination. All main components exhibit Auger valence band transitions which change their shape with the chemical state. Chemometric methods have been utilized to identify characteristic peak shapes and to classify the investigated areas by composition. Fibre fracture surfaces are characterized by Al, magnesia and Li oxide formed by a redox reaction of alumina with Mg and Li penetrating along grain boundaries. For samples with high silica content a thin interfacial oxide layer on matrix grain boundaries and matrix-fibre interfaces has been found. 相似文献
16.
17.
Summary Modified nucleosides derived predominantly from transfer ribonucleic acid (tRNA) have been studied as possible tumor markers.
In this paper a reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been applied to study 15 normal
and modified nucleosides in serum. The nucleoside levels in normal human serum were established, and the concentrations of
15 nucleosides in serum from 19 cancer patients were determined. It was found that the concentrations of modified nucleosides
were significantly higher in patient sera. Based on 15 nucleoside concentrations in serum, factor analysis could classify
correctly 90% of cancer patients from the normal humans. Further work showed that urine was slightly better than serum when
determining nucleosides as biological marker candidates. More work is ongoing to determine the clinical usefulness of modified
nucleosides as tumor markers. 相似文献
18.
19.
细胞中鸟苷二磷酸糖的毛细管电泳研究 总被引:1,自引:0,他引:1
葡萄糖是细胞生长和代谢的一种重要的调节剂 .鸟苷二磷酸糖 (UDP Sug ars)作为其代谢的中间产物 ,在碳水化合物、糖类酯等的合成中起着重要作用 .研究其在细胞中的代谢在临床生物化学中具有重要意义 .建立了一个毛细管电泳方法 ,采用 70cm× 5 0 μm的未涂毛细管 ,2 0mmol/L硼酸盐 (pH9)作缓冲液 ,在 2 2kV下 ,可在15min左右全分离 4个关键UDP Sugars,精度和检测限满足细胞分析要求 .经实际分析淋巴细胞、成纤维细胞和肾小球细胞 ,表明此方法不仅比HPLC具有明显优点 ,而且还可用于测量细胞能荷 相似文献
20.
Hydroxycarboxylic and oxocarboxylic acids in urine: products from branched-chain amino acid degradation and from ketogenesis 总被引:2,自引:0,他引:2
Hydroxy- and oxomonocarboxylic acids in urine of healthy individuals and of patients with diabetic ketoacidosis are analysed as methyl esters and methyl esters/O-methyloximes, respectively, by gas chromatography and gas chromatography-mass spectrometry. The derivatives are pre-fractionated by thin-layer chromatography. The acids originate mainly from ketogenesis and from the metabolism of valine, leucine and isoleucine. The amino acid metabolites fall into three groups: the 2-oxocarboxylic acids (2-oxoisovaleric acid, 2-oxoisocaproic acid and 2-oxo-3-methylvaleric acid); the 2-hydroxycarboxylic acids (2-hydroxyisovaleric acid, 2-hydroxyisocaproic acid and 2-hydroxy-3-methylvaleric acid); and the 3-hydroxycarboxylic acids (3-hydroxyisobutyric acid, 3-hydroxyisovaleric acid, 3-hydroxy-2-ethylpropionic acid, threo-3-hydroxy-2-methylbutyric acid and erythro-3-hydroxy-2-methylbutyric acid). The threo form of 3-hydroxy-2-methylbutyric acid is the major constituent within the diastereomeric pair. Of the three groups of amino acid metabolites, the 3-hydroxycarboxylic acids in particular are elevated during ketoacidosis. The characteristic general features of the mass spectrometric fragmentation of the derivatives of the identified components are systematically described. The discussion of the fragmentation includes constituents of low concentrations, such as 3-oxocaproic acid, 4-oxobutyric acid and 5-oxocaproic acid, which can be detected only when the pre-fractionation technique is applied. 相似文献