Semi-automatic liquid chromatographic analysis of pamidronate in serum and citrate plasma after derivatization with 1-naphthylisothiocyanate.
The semi-automatic method for the determination of the bisphosphonate pamidronate in serum and citrate plasma involves a manual protein precipitation with trichloroacetic acid and a manual coprecipitation of the bisphosphonate with calcium phosphate, followed by an automated solid-phase extraction on anion-exchange columns. After off-line evaporation of the extract under nitrogen and reconstitution in water, the automatic procedure is continued by automatic derivatization with 1-naphthylisothiocyanate, ion-pair liquid-liquid extraction and a treatment with hydrogen peroxide, prior to analysis by ion-pair HPLC and fluorescence detection at 285/390 nm. The intra- and inter-day precisions are 1.3 and 7%, respectively, for a standard of 100 ng ml(-1) pamidronate in serum; the average accuracy for this standard is 107%. The lower limit of quantification is 20 ng ml(-1) pamidronate in 1 ml of human serum.
Sparidans
R W
RW
Department of Clinical Pharmacy and Toxicology, Leiden University Medical Centre, The Netherlands.
den Hartigh
J
J
Beijnen
J H
JH
Vermeij
P
P
D015058:1-Naphthylisothiocyanate / Q000032:analysis*; D000368:Aged; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D019343:Citric Acid / Q000097:blood*; D004164:Diphosphonates / Q000097:blood*; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood; D000077268:Pamidronate; D012016:Reference Values; D015203:Reproducibility of Results; D012371:Robotics; D012680:Sensitivity and Specificity; D013455:Sulfur / Q000378:metabolism
D004164:Diphosphonates; D019343:Citric Acid; D015058:1-Naphthylisothiocyanate; D013455:Sulfur; D006861:Hydrogen Peroxide; D000077268:Pamidronate