Sensitive determination of a novel bisphosphonate, YM529, in plasma, urine and bone by high-performance liquid chromatography with fluorescence detection.
A high-performance liquid chromatographic method for the sensitive determination of 1-hydroxy-2-(imidazo[1,2-a]pyridin-3-yl)ethane-1, 1-bisphosphonic acid monohydrate (YM529) in plasma, urine and bone is described. Plasma obtained in high-dose animal studies is treated by method A, a simple method using 1 ml of plasma, which is based on deproteinization of plasma followed by coprecipitation of the drug with calcium phosphate and dissolution of the precipitate in EDTA. Plasma obtained in low-dose clinical studies is treated by method B, a more sensitive method using 4 ml of plasma, which is based on direct precipitation of the drug prior to the deproteinization in method A. Urine and bone samples are prepared by solid-phase extraction using a Sep-Pak C18 cartridge coupled with method A. The drug is separated with a reversed-phase column using a mobile phase at pH 7, and detected with a fluorescence detector following postcolumn alkalization of the mobile phase to enhance fluorescence intensity. The limit of determination is 0.2 ng/ml for method A and 0.05 ng/ml for method B in plasma, 0.05 ng/ml in urine, and 5 ng/g in bone.
Usui
T
T
Drug Metabolism Department, Yamanouchi Pharmaceutical Co., Tokyo, Japan.
Kawakami
R
R
Watanabe
T
T
Higuchi
S
S
D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D002130:Calcium Phosphates; D002851:Chromatography, High Pressure Liquid; D004164:Diphosphonates / Q000032:analysis* / Q000097:blood / Q000652:urine; D004285:Dogs; D004492:Edetic Acid; D006801:Humans; D007093:Imidazoles / Q000032:analysis* / Q000097:blood / Q000652:urine; D007202:Indicators and Reagents; D051381:Rats; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D002130:Calcium Phosphates; D004164:Diphosphonates; D007093:Imidazoles; D007202:Indicators and Reagents; C087958:YM 529; D004492:Edetic Acid