Determination of alendronate in low volumes of plasma by column-switching high-performance liquid chromatography method and its application to pharmacokinetic studies in human plasma

• Published in: Archives of pharmacal research Vol. 34; no. 12; pp. 2079 - 2086
• Main Authors: Ban, Eunmi, Park, Ji-Youn, Kim, Hyung Tae, Kim, Chong-Kook
• Format: Journal Article
• Language: English
• Published: Heidelberg Pharmaceutical Society of Korea 01.12.2011; 대한약학회
• Subjects: Alendronate - blood; Alendronate - pharmacokinetics; Chemistry, Pharmaceutical - methods; Chemistry, Pharmaceutical - standards; Chromatography, High Pressure Liquid - methods; Chromatography, High Pressure Liquid - standards; Humans; Medicine; Pharmacology/Toxicology; Pharmacy; Reproducibility of Results; 약학; High-performance liquid chromatography (HPLC); Column switching; Pharmacokinetic study; Alendronate
• ISSN: 0253-6269; 1976-3786
• DOI: 10.1007/s12272-011-1211-z

A sensitive and simple column-switching high-performance liquid chromatographic (HPLC) method with fluorescence detection was developed for the determination of alendronate in human plasma. Alendronate and pamidronate (internal standard, IS) in plasma including Na 2 EDTA were precipitated with sodium hydroxide and calcium chloride after deproteinization using 10% trichloroacetic acid solution. The precipitated alendronate and IS were reconstituted by sodium citrate and citric acid and then derivatized with 9-fluorenylmethyl chloroformate. The resulting solution was injected onto an HPLC system consisting of a pretreatment column and an analytical column, which were connected with a six-port switching valve. The assay was linear in the concentration range of 2–100 ng/mL in 1 mL of plasma with high precision and accuracy, and the limit of detection was 0.5 ng/mL. It was successfully applied to evaluate the pharmacokinetic parameters of alendronate in human volunteers following single oral administration. The mean value of maximum alendronate plasma concentration (C max ) was 37.69 ng/mL, and the mean time to reach the C max (T max ) was 1.08 h. The area under the plasma concentration-time curve (AUC) and elimination half-life (T 1/2 ) were 106.48 ng/mL/h and 1.66 h, respectively.