Quantitation of Amino Acids by Gas-Liquid Chromatography

摘要: Summary The GC analysis of amino acids such as the N-trifluoroacetyl (N-TFA) n-butyl esters – established method developed principally in our laboratories provides an effective and reliable means acid determination that is applicable to a very wide range analytical needs. My research group, graduate students, colleagues during period from 1960 1975, fundamentals quantitative derivatization, conditions chromatographic separation, defined interactions derivatives with stationary support phases. Our studies continued refinements since 1974 have resulted precise accurate, reliable, straightforward for measurement [ [1] , [2] [3] [4] ]. We conducted extensive array applications on sample matrices, pine needle extracts erythrocytes. Experimental section (Volume 1, Chapter 1 ) 2 ] thorough description procedures, including preparation ethylene glycol adipate (EGA) silicone-mixed phase columns. EGA column which used separate quantitate all protein acids, except histidine, arginine, cystine composed 0.65 w/w% stabilized grade 80/100 mesh acid-washed Chromosorb ® W, 1.5 m × 4 mm ID glass. For quantitation cystine, 1.0 OV-7 0.75 SP-2401 100/120 Gas-Chrom® Q (1.5 I.D. glass) performs extremely well. also describe use ion-exchange resins cleanup, complete derivatization N-TFA ≪-butyl esters. are esterified by reaction rc-butanol • 3 N HC1 15 min at 100°C excess ≪-butanol’ removed under vacuum 60°C any remaining moisture azeotropically dichloromethane; then trifluoroacylated trifluoroacetic anhydride (TFAA) 150° C 5 presence dichloromethane solvent. Immediately following valuable comments various parts ], provide guidance entire technique, chromatography quantitation. Of particular value comparison GLC IEC results hydrolysates diverse matrices. This types showed values obtained two techniques were generally close agreement. N-'IFA ≫-butyl technique offers much scientists concerned acids. excellent precision, accuracy, selectivity, economical complementary elegant Stein-Moore method. both detailed account historical perspective development solution problems encountered methods evolved w-butyl ester trimethylsilyl (TMS) discussed, conditions, separations, mass spectrometric (MS) identification classes derivatives, liquid materials, application acylation arginine posed problem early studies; successful this paved way high-temperature procedure, now widely numerous acylating reagents [5] [6] Likewise, esterification was investigated detail, resulting direct procedure quickly reproducibly converts approach has been form iodine- sulfur-containing TMS described, finding bis (trimethylsilyl)trifluoroacetamide (BSTFA), silylating reagent we invented patented, silylation reagent, BSTFA, led conversion volatile single step. Although certain tend multiple contain varying numbers groups, high-temperature, long time permits derivatives. 6 10% OV-11 Supelcoport® separation system VV-TFA came about realization not eluted columns polyester phases, although type other siloxane mixed specifically these three final being 17 summary points out, foundation elements: (a) reproducible suitable (b) elution column. A literature review presented 1984 2005 topics ranging N-acyl O-esters to: Enantiomeric composition, racemization, gas enantiomer formation cyclosporine metabolites, 1- 3-methyl histidine biological fluids, rapid analyses, resolution chiral columns, simultaneous functional groups one-step ethyl chloroformate supercritical CO2, diasteromeric a-alkyl- a-amino capillary plus subjects (see titles authors follows).