Lamotrigine Extraction and Quantification by UPLC-DAD in Plasma from Patients with Bipolar Disorder

Abstract

A sensitive and efficient analytical process for detecting lamotrigine in acidic solution based in ultra-high-performance liquid chromatography-diode array detector (UPLC-DAD) was developed; the stationary phase used was a C8, 150 × 4.6 mm, 2.6 µm. The mobile phase consisted of acetonitrile/acidified water (0.01% H3PO4 and 0.005% triethylamine, pH 2.4) (25 : 75 v/v). Limits of detection and quantification were 0.02 µg/mL and 0.05 µg/mL, respectively. The working interval for the evaluation of the method ranged from 0.05 to 12 µg/mL, and the linear fit of the experimental data has a value of r2≥0.98. Before quantifying lamotrigine in plasma of patients with bipolar disorder, lamotrigine was released from plasma proteins with a 0.2 M sodium hydroxide solution, and then proteins were removed by precipitation with acetonitrile. Afterward, the lamotrigine base was dissolved in ethyl acetate. This extract was reconstituted in potassium phosphate solution (pH 2.4) to obtain more than 98% of lamotrigine protonated in N2, which was detected and quantified as indicated above. The absolute percentage of lamotrigine recovery is ≥80% for all tested concentration levels. The accuracy and precision of the method have %CV values <4% for the lamotrigine levels of 3, 6, and 9 µg/mL. The correlation coefficient for the used concentration range is 0.99. The analytical method is precise and sensitive to measure lamotrigine levels expected in plasma of BD patients and these levels were in the therapeutic dose range.