A GAS-LIQUID-CHROMATOGRAPHIC PROCEDURE FOR SEPARATING A WIDE RANGE OF METABOLITES OCCURRING IN URINE OR TISSUE EXTRACTS
A GAS-LIQUID-CHROMATOGRAPHIC PROCEDURE FOR SEPARATING A WIDE RANGE OF METABOLITES OCCURRING IN URINE OR TISSUE EXTRACTS
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DOI：
10.1042/bj1010792
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发表时间：
1966-01-01
影响因子：
4.1
通讯作者：
YARGER, K
中科院分区：
文献类型：
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作者：
DALGLIESH, CE;HORNING, EC;YARGER, K
1.描述了一种允许在同一色谱图上分离和鉴定出现在尿液或组织提取物中的各种物质的气-液-色谱程序。该方法使用氢火焰电离，检测有机化合物，无论是自由的还是共轭的，而不需要特定的反应基团。2.对于层析，羧基被定量地转化为甲酯或三甲基硅酯。酚基、醇基和潜在的烯基转化为三甲基硅基醚。分离是在一台6英尺长的飞机上进行的。色谱柱为10%F-60(聚硅氧烷)或1%F-60，程序升温2度/分钟。根据需要在30度-260度的温度范围内使用。羟基化合物的丙酰基衍生物也可以使用，但只能在非定量的基础上使用。当使用自动气相色谱检查大量样品时，已选择了最佳使用范围的衍生物和色谱柱。3.本方法适用于：丁酸以上的脂肪酸；二和三羧酸；羟基酸和酮酸；多羟基和脂环化合物，如甘油、肌醇、奎尼酸、莽草酸、抗坏血酸和糖醇；芳香族羟基和酸性化合物，苯类和吲哚类；倍半萜；类固醇；甘氨酸共轭化合物；硫代尿酸；葡萄糖醛酸类。硫酸盐偶联物、咪唑类化合物或多肽的应用效果不理想。4.亚甲基单元为表征峰位置提供了准确和可重复性的参数。据报道，尿液和组织萃取物中存在的或与之相关的各种物质的亚甲基单位值。5.采用气相色谱-质谱法相结合的方法对化合物的性质进行了确认。气相色谱-质谱联用为代谢模式的评价提供了一种强有力的诊断工具，并讨论了各种用途。
1. A gas-liquid-chromatographic procedure is described which permits separation and identification on the same chromatogram of a wide range of substances occurring in urine or tissue extracts. The method uses hydrogen flame ionization, which detects organic compounds whether free or conjugated with no requirement for specific reactive groups. 2. For chromatography, carboxyl groups are quantitatively converted into methyl esters or trimethylsilyl esters. Phenolic, alcoholic and potential enolic groups are converted into trimethylsilyl ethers. Separations are carried out on a 6ft. column of either 10% F-60 (a polysiloxane) or 1% F-60, temperature programming at 2 degrees /min. being used over such part of the temperature range 30 degrees -260 degrees as is required. Propionyl derivatives of hydroxy compounds can also be used, but only on a non-quantitative basis. Derivatives and columns have been selected for optimum range of usefulness when large numbers of samples are examined by using automated gas chromatography. 3. The method is applicable to: fatty acids above butyric acid; di- and tri-carboxylic acids; hydroxy acids and keto acids; polyhydroxy and alicyclic compounds such as glycerol, inositol, quinic acid, shikimic acid, ascorbic acid and sugar alcohols; aromatic hydroxy and acidic compounds, both benzenoid and indolic; sesquiterpenes; steroids; glycine conjugates; mercapturic acids; glucuronides. It is not satisfactory for sulphate conjugates, iminazoles or polypeptides. 4. Methylene units provide an accurate and reproducible parameter for characterizing peak position. Methylene unit values are reported for a large variety of substances occurring in, or related to those occurring in, urine and tissue extracts. 5. The nature of derivatives was confirmed by combining gas chromatography with mass spectrometry. Combined gas chromatography-mass spectrometry gives a diagnostic tool of great power in the evaluation of metabolic patterns, and various uses are discussed.