Computer analysis of amino acid chromatograms.

A procedure is described for the automatic off-line analysis of amino acid chromatograms of protein hydrolysates, using a small computer. The data requirements are basic, and, unlike previous programs, the present system allows the separation and identification of bands, as well as the quantitative determination of composition. With minor modification, the program could be extended for use with most types of chromatographic data. The validity of the application of the program to experimental data is discussed.     

Fluorescent oxidation products derived from DBD-thiocarbamoyl amino acid in the modified Edman sequencing analysis

The fluorescent product obtained by the oxidation of 7-N, N-dimethylaminosulfonyl-4-(2,1,3-benzoxadiazolyl) (DBD)-thiocarbamoyl (TC)-proline with NaNO(2)/H(+) in the modified Edman sequencing procedure was identified as the corresponding thiazolyl compound, N-[(8-dimethylaminosulfonyl)thiazolo[5,4, e]benzo[2,1,3]oxadiazol-5-yl]-L-proline, formed by the attack of the sulfur atom of the thiocarbamoyl group on the benzofurazan skeleton. The reaction mechanism for the formation of the fluorescent compound from DBD-TC primary and secondary amines is also discussed.     

Applications of automated amino acid analysis using 9-fluorenylmethyl chloroformate.

A rapid and sensitive fully automated method for the determination of primary and secondary amino acids in different matrices is described. Amino acids are derivatized with 9-fluorenylmethyl chloroformate using an automated precolumn derivatization technique. Data are presented to show that the technique is both reproducible and highly sensitive. Applications of the technique are presented, including the analysis of peptide and protein hydrolysates and the profiling of free amino acids in physiological fluids.     

Computer analysis of amino acid chromatograms and other linear elution spectra

A computer-aided method of evaluation of amino acid column chromatograms is proposed and compared with the usual synchronous peak-integration method. Spectra and their backgrounds are digitized separately by a curve digitizer; this allows a better estimate of the background time courses. A computer program then fits a set of superimposed Gaussian distributions to each corrected spectrum, thus circumventing the problems arising from incompletely separated peaks. Samples of known composition, run intermittently through the analyzer between the unknown spectra, allow a determination of the time dependence of the ratios of “peak areas over amounts of amino acids”; hence, the amounts of amino acids can automatically be corrected for aging effects of the analyzing system. A modified version of the computer program allows resolution of any spectrum to a sum of Gaussians, with least squares fitting of their amplitudes, widths and locations.     

Multivariate 3-way data analysis of amino acid patterns of lakes

Time dependent patterns of amino acid concentrations have been studied by HPLC for different lakes of the Berlin area. Data analysis has been performed by conventional principal component analysis as well as by its more recent N-way extension. It turns out that lakes mainly differ by their general amino acid production as a function of time and season. Apart from this, in a single case there occurs a specific pattern which might be related to an exterior influence. This pattern, although clearly detected, has been not stable over time. Measurements are reproducible with respect to time (comparison of two succeeding years) and to position (comparison of isolated parts of a lake).Dedicated to Professor Dr. K. Doerffel at his 70th birthday with respect to his fundamental contributions to chemometrics     

Peptide separation and isolation with an automated amino acid analyzer

A technique is presented for separation and collection of amino acids and peptides using a microcolumn amino acid analyzer. By use of the program and the column selection valve of the amino acid analyzer, and without any modification of the instrumentation, the stream of the eluate is diverted into the reaction coil and absorbance is recorded at regular intervals. The rest of the eluate is collected in a fraction collector for further characterization of the separated peptides. Splitting ratios can be varied by simple alteration of the program. Small amounts of material (1 to 2 nmol) are needed for monitoring the separation and collection of the eluate.     

Computer simulation of amino acid sorption on carbon nanotubes

A computer simulation of complexes of (6,6) open carbon nanotubes (CNTs) with neutral molecules, zwitterions and glycine, alanine, and phenylalanine amino acid anions is performed. In starting structures amino acids are arranged in three types: on the external side face, the open end, and inside CNT. The structure is optimized within the density functional theory with regard to the GD3 dispersion correction with and without taking into account solvation effects. It is found that the greatest CNT–amino acid interaction occurs in the neutral aqueous medium at dissociative chemisorption of the zwitterion (adsorption energy 80-90 kcal/mol) and in the basic medium at anion chemisorption (energy ~48-50 kcal/mol) on the open CNT end.     

An automated activation analysis data acquisition system

An automated neutron activation analysis data acquisition system has been assembled from commercially available equipment. The modifications of the components needed to make this into a working system are described in the text. The main components of the data acquisition system are a sample changer, a Ge(Li) detector, a magnetic tape deck and a minicomputer based multichannel analyzer. The sample changer has a 200-sample capacity and can handle both solid and liquid samples. Software for controlling the data acquisition system is flexible, yet simple to use. The system has operated reliably for a year and has sharply reduced the effort needed for data acquisition.     

Microfluidics in amino acid analysis

Microfluidic devices have been widely used to derivatize, separate, and detect amino acids employing many different strategies. Virtually zero-dead volume interconnections and fast mass transfer in small volume microchannels enable dramatic increases in on-chip derivatization reaction speed, while only minute amounts of sample and reagent are needed. Due to short channel path, fast subsecond separations can be carried out. With sophisticated miniaturized detectors, the whole analytical process can be integrated on one platform. This article reviews developments of lab-on-chip technology in amino acid analysis, it shows important design features such as sample preconcentration, precolumn and postcolumn amino acid derivatization, and unlabeled and labeled amino acid detection with focus on advanced designs. The review also describes important biomedical and space exploration applications of amino acid analysis on microfluidic devices.     

Advances in amino acid analysis

Amino acids are important targets for metabolic profiling. For decades, amino acid analysis has been accomplished by either cation-exchange or reversed-phase liquid chromatography coupled to UV absorbance or fluorescence detection of pre-column or post-column-derivatized amino acids. Recent years have seen great progress in the development of direct-infusion or hyphenated mass spectrometry in the analysis of free amino acids in physiological fluids, because mass spectrometry not only matches optical detection in sensitivity, but also offers superior selectivity. The advent of cryo-probes has also brought NMR spectroscopy within the detection limits required for the analysis of free amino acids. But there is still room for further improvement, including expansion of the analyte spectrum, reduction of sample preparation and analysis time, automation, and synthesis of affordable isotope standards. Figure Fully automated gas chromatography-mass spectrometry analysis of amino acids.