Determination of amino acids by gas chromatography using a wide bore capillary column

A procedure is described in which a wide bore capillary column is used as an alternative to the more traditional packed column for the quantitative analysis of amino acids as their N-heptafluorobutyryl isobutyl ester (HBB) derivatives. The column, installed in a gas chromatograph previously configured for use with a packed column, is shown to give good reproducibility by repeated determination of amino acid response factors (RSD values for all amino acids are below 3%). A number of problems, encountered during the use of this column, are discussed and suitable techniques to overcome them are reported.     

Fundamentals and applications of ion-ion plasmas

Ion-ion plasmas can form in the late afterglow of pulsed discharges or downstream of continuous wave discharges in electronegative gases. In ion-ion plasmas, negative ions replace electrons as the negative charge carriers. In the absence of electrons, ion-ion plasmas behave quite differently compared to conventional electron-ion plasmas. Application of a radio frequency bias to a substrate immersed in an ion-ion plasma can be used to extract alternately positive and negative ions, thereby minimizing charging on device features during micro-device fabrication. Ion-ion plasmas are also important in negative ion sources, dusty plasmas, and the D-layer of the earth's atmosphere.     

A review of quantitative ion exchange,high performance liquid and gas chromatographic analysis of amino acids in physiological fluids

Methodologies for quantitative analysis of amino acids in physiological fluids based on classical ion exchange, high performance liquid chromatography, and gas chromatography are analyzed. As judged by the relative number of reports on these techniques, it appears that classical ion exchange continues to be the main method of choice for amino acid determinations and only limited advantage has been taken of the benefits (e.g. lower capital outlay and running costs and shorter analysis times) offered by other techniques. More importantly, however, there appears to be insufficient quantitative evaluation of the methodologies used. As a result, the validity of many reports based on data from amino acid analysis of physiological samples may be questioned.     

Determination of amino acids in the hepatic and brain tissue of the rat by gas chromatography

A procedure is described for the quantitative determination of amino acids in hepatic and brain tissue samples from the rat. Because the presence of certain matrix components in the tissue material led to interference with chromatographic analysis they were removed by a prechromatographic “clean-up” step. Quantitative analysis of amino acids, as their N-heptafluorobutyryl iso-butyl ester derivatives, was achieved by high resolution gas chromatography on an apolar fused silica open tubular column. Reproducibility data from the complete procedure are presented; coefficients of variation for arginine and histidine in hepatic tissue varied between 7.1 and 10.1% whereas those for most other amino acids were better than 5%, with a mean recovery of 90%.     

Gas chromatography of plasma amino acids using a wide bore column: Comparison with packed and capillary columns

A procedure is described in which a wide bore glass capillary column is used as an alternative to the more traditional packed column in the analysis of amino acid levels in plasma. The coefficients of variation for all amino acids (with the exception of aspartic acid) were better than 11% with recoveries ranging from 81% to 122%. The data are compared with the corresponding results obtained using a packed column and show significant differences (p < 0.005) between values for glycine, serine, isoleucine, proline, methionine, aspartic acid, phenylalanine, and lysine. A similar comparison between results from the wide bore and the fused-silica open tubular (FSOT) column shows better agreement. Adjustment of chromatographic conditions for the wide bore analysis yields results in good agreement with those from FSOT analysis but which still differe significantly from the backed column data.     

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).     

Determination of amino acids in human cerebrospinal fluid by gas chromatography

In the present study, the use of gas chromatography (GC) for the determination of amino acids in human cerebrospinal fluid (CSF) is described. Although some amino acids may be determined using a packed column following the removal of glucose, the major interfering component, the inadequate resolution of other amino acids from remaining unidentified components results in poor quantitation. The use of wide bore columns improves reproducibility considerably, but still it does not provide sufficient resolution to enable quantitative determination of all amino acids in human CSF. Good reproducibility data, with CV values for all amino acids of 7% or less and recoveries generally between 80% and 100%, can only be obtained using the fused silica open tubular (FSOT) column. Normal amino acid levels are presented for 10 samples of human CSF, which compare well with data previously reported in the literature.     

Determination of pentane in human breath by gas chromatography

A new procedure, requiring expired sample volumes of approximately 2L, has been developed for the quantitative determination of n-pentane in human breath. The methodology capitalizes on the superior resolving power and separating efficiency of the open tubular capillary column and incorporates an effective prechromatographic cryofocusing technique. Molecular sieve is used to simultaneously remove moisture and carbon dioxide from breath samples which are “spiked” with an internal standard to optimize the validity of analysis. Reproducible data for both internal standard spiking (CV = 5.8%) and sample n-pentane level (CV = 3.2%) have been obtained, together with a mean recovery of 104%. Preliminary data show n-pentane levels in normal female subjects (n = 10) to range from 0.62 to 3.16 nmole/L (mean 1.76 nmole/L) and in males (n = 10) from 3.20 to 8.76 nmole/L (mean 5.66 nmole/L).