A thorough LC-MS/MS strategy to quantify abaloparatide in rat plasma: method development, validation and application

Area of Science:

• Analytical Chemistry
• Pharmacology
• Biotechnology

Background:

• Abaloparatide is a parathyroid hormone-related peptide analog for osteoporosis treatment in postmenopausal women.
• High fracture risk necessitates effective osteoporosis management strategies.
• Accurate quantification of abaloparatide in biological samples is crucial for its development and clinical application.

Purpose of the Study:

• To develop a novel, rapid, sensitive, and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for abaloparatide quantification in plasma.
• To validate the developed LC-MS/MS method for accuracy, precision, and reliability.
• To assess the stability of abaloparatide in plasma under various storage conditions.

Main Methods:

• Plasma samples were prepared using protein precipitation.
• Abaloparatide was separated using a positively charged surface hybrid column.
• Quantification was performed using multiple-reaction mass spectrometry detection with selected ion transitions.
• Method validation included accuracy, precision, recovery, matrix effect, and stability assessments.

Main Results:

• The LC-MS/MS method demonstrated high sensitivity, quantifying abaloparatide in the range of 0.02 to 8 ng/mL.
• The method exhibited excellent accuracy and precision, with acceptable recoveries for abaloparatide and the internal standard.
• Abaloparatide stability was confirmed under various storage conditions, including 4°C, -80°C, and freeze-thaw cycles.
• The method was successfully applied to a pharmacokinetic study in rats.

Conclusions:

• This is the first mass spectrometry-based method for quantifying abaloparatide in plasma.
• The developed LC-MS/MS method provides a reliable and sensitive tool for abaloparatide quantification.
• This method may facilitate abaloparatide development and advance the application of LC-MS/MS in peptide bioanalysis.