High-performance liquid chromatographic determination of 6-aminopenicillanic acid by postcolumn alkaline degradation.
A high-performance liquid chromatographic method has been developed for the determination of 6-aminopenicillanic acid in amino acid mixtures and human serum. The separation of 6-aminopenicillanic acid was carried out on a C18 column using sodium heptylsulfonate or tetrabutylammonium bromide as an ion-pairing agent and methanol as an organic mobile phase modifier. Detection was based on a postcolumn reaction with sodium hydroxide, mercury(II) chloride, and ethylenediaminetetraacetic acid disodium salt followed by measurement of ultraviolet absorbance (at 300 nm) of the reaction product(s). The method is quantitative for 6-aminopenicillanic acid concentrations down to 0.1 microgram/ml in human serum samples with a 20-microliter injection. At a concentration of 2 micrograms/ml, the within- and between-run precisions (relative standard deviation) were 1.29-3.91% and 2.30%, respectively.
D000596:Amino Acids / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D010397:Penicillanic Acid / Q000032:analysis* / Q000097:blood / Q000031:analogs & derivatives; D012972:Sodium Hydroxide; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D012972:Sodium Hydroxide; D010397:Penicillanic Acid; C004408:aminopenicillanic acid