Ultramicro GC Determination of Amino Acids Using Glass Open Tubular Columns and a Nitrogen-Selective Detector

Abstract

A glass open tubular (capillary) column together with a nitrogen- selective detector were evaluated for the analysis of amino acids by gas chromatography. Test samples included normal and abnormal human sera and urine and several hydrolysates of pure proteins. Amino acids in the biological samples were isolated by ion exchange pretreatment and derivatized to the n-propyl, N-acetyl derivatives. Protein hydrolysates were derivatized without pretreatment. Compared with a flame ionization detector, the nitrogenselective detector gave a sensitivity enhancement of 80x – 180x for the amino acid derivatives, depending on the particular amino acid. Long-term stability of the detector was excellent. Reproducibility studies gave a long-term precision of about 6% for serum or urine samples while a precision of 5% was obtainable on a day-to-day basis when analyzing protein hydrolyzates. The experiments showed that amino acids at the picomole level could be reliably analyzed with the system. The open tubular column coated with a mixture of Carbowax 20M and Silar 5 CP and having a very thin liquid phase film gave greatly improved thermal stability compared with a previously used packed column.