A simple method to detect contaminating peptides and amino acids in large-scale ganglioside preparations.
A reverse-phase liquid chromatography method was developed to analyse the presence of contaminating peptides and amino acids in large-scale monosialoganglioside preparations. Samples were hydrolysed under controlled conditions and derivatized with phenylisothiocyanate. PTC-amino acids were then separated and identified by HPLC. The sensitivity of the method allowed detection of a least 50 pmoles of amino acid per mg of ganglioside with excellent reproducibility and little or no interference of by-products derived from hydrolysis of the glycosphingolipid. The response was consistently linear for each amino acid within the examined analytical range. The ease and speed of performance make this method a useful tool for rapidly monitoring large-scale extraction and purification processes of biologicals obtained from natural sources.
Corona
G
G
Advanced Technology Division, Fidia S.p.A., Abano Terme, Italy.
Mattioli
S
S
Di Martino
A
A
Callegaro
L
L
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004340:Drug Contamination; D005677:G(M1) Ganglioside / Q000302:isolation & purification*; D017879:Isothiocyanates; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013861:Thiocyanates
D000596:Amino Acids; D017879:Isothiocyanates; D009842:Oligopeptides; D013861:Thiocyanates; C005441:phenylisothiocyanate; D005677:G(M1) Ganglioside