Nateglinide quantification in rabbit plasma by HPLC: optimization and application to pharmacokinetic study

Area of Science:

• Analytical Chemistry
• Pharmacokinetics
• Biomedical Science

Background:

• Nateglinide (NTG) is an antidiabetic drug requiring precise pharmacokinetic analysis.
• Existing methods for NTG quantification in biological matrices may lack speed, simplicity, or sensitivity.
• Validated analytical methods are essential for drug development and therapeutic drug monitoring.

Purpose of the Study:

• To develop and validate a rapid, simple, and sensitive High-Performance Liquid Chromatography (HPLC) method for nateglinide (NTG) determination in rabbit plasma.
• To investigate the method's performance characteristics, including linearity, sensitivity, precision, and accuracy.
• To apply the validated method for pharmacokinetic studies of NTG in rabbits.

Main Methods:

• Development of an HPLC method using a C18 column with UV detection at 203 nm.
• Optimization of mobile phase composition (acetonitrile:phosphate buffer, pH 3.0) and flow rate (1.0 ml/min).
• Plasma sample preparation via deproteinization with methanol/acetonitrile mixture, followed by quantification using drug/internal standard peak area ratios.

Main Results:

• The method achieved a run time of under 8 minutes with high extraction efficiency (>97.86%).
• Linearity was demonstrated over a wide concentration range (10-2500 ng/ml) with a high correlation coefficient (0.9984).
• Excellent sensitivity (LoD: 2.91 ng/ml, LoQ: 9.70 ng/ml) and precision (R.S.D. <1.32%) were achieved.

Conclusions:

• The developed HPLC method is rapid, simple, sensitive, and selective for NTG determination in rabbit plasma.
• The method eliminates the need for tedious sample clean-up procedures.
• The validated assay is suitable for pharmacokinetic profiling of nateglinide in preclinical studies.