Extraction and measurement of pamidronate from bone samples using automated pre-column derivatization, high-performance liquid chromatography and fluorescence detection.
A method for the determination of 3-amino-1-hydroxylpropylidene-1, 1-bisphosphonic acid (pamidronate) in bone samples is described. This method combines and modifies parts of previous procedures. Pamidronate is extracted from finely ground bone with dilute hydrochloric acid. Amine-containing contaminants are removed by co-precipitation of pamidronate with calcium. Excess calcium is removed with EDTA and an ion-exchange resin. Pamidronate is automatically derivatized at the primary amine and quantified by high-performance liquid chromatography with fluorescence detection. The method assay was linear in the concentration range 7.5-600 ng/mg bone (20-100 pmol/mg). The imprecision for repeat analyses were 16.5 and 7.8%, at pamidronate levels of 7.5 and 600 ng/mg bone, respectively. The method has been used to analyze bone samples from pharmacokinetic animal studies involving both acute and chronic dosages.
King
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Department of Clinical Biochemistry, University of Toronto, Ont., Canada.
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D000818:Animals; D001322:Autoanalysis; D001842:Bone and Bones / Q000737:chemistry*; D002118:Calcium; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004164:Diphosphonates / Q000032:analysis* / Q000493:pharmacokinetics; D004285:Dogs; D004492:Edetic Acid; D006851:Hydrochloric Acid; D007202:Indicators and Reagents; D000077268:Pamidronate; D012680:Sensitivity and Specificity
D004164:Diphosphonates; D007202:Indicators and Reagents; D004492:Edetic Acid; D000077268:Pamidronate; D006851:Hydrochloric Acid; D002118:Calcium