SENSITIVE AND SPECIFIC LC-MS/MS METHOD FOR THE SIMULTANEOUS DETERMINATION OF CHLORPROGUANIL, DAPSONE, AND THEIR METABOLITES IN HUMAN PLASMA

Abstract

A sensitive, specific, and rapid liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for the determination of dapsone, chlorproguanil, and their metabolites was developed and validated over a concentration range of 2–2000 ng/mL using 200 µL of plasma. After a simple solvent precipitation procedure, the supernatant was analyzed directly by HPLC-MS/MS method using an XTerra RP18 (2.1 mm × 100 mm, 5.0 µm) column with mobile phase consisting of acetonitrile-0.1% formic acid in water (75:25, v/v, pH 3.0) in an isocratic mode at a flow rate of 0.3 mL/min. Mass detection was performed using a triple quadrupole mass spectrometer operating in positive electrospray ionization mode. The elution of chlorproguanil (CPG, 288 ->204), chlorcycloguanil (CCG, 286 ->229), dapsone (DDS, 249 ->156), monoacetyldapsone (MADDS, 291 ->156), and trimipramine-D₃ (TMP-D₃, 298 ->103) was monitored using multiple reaction monitoring. The lower limit of detection for CPG, CCG, DDS, and MADDS was 0.5 ng mL⁻¹ while the limit of quantification was 2 ng mL⁻¹ in human plasma. The extraction recovery of CPG, CCG, DDS, MADDS, and TMP-D₃ from human plasma was higher than 87%. The method may find its application in therapeutic monitoring of these compounds in biological matrix.

Keywords:

• chlorcycloguanil
• chlorproguanil
• dapsone
• HPLC-MS/MS
• monoacetyl dapsone
• therapeutic monitoring

ACKNOWLEDGMENT

The authors sincerely acknowledge the financial support from Uttarakhand State Council for Science and Technology, Uttarakhand to perform this study.

Notes

*Precision [RSD (%)] = (SD/mean) × 100.

**Accuracy (%) = (mean measured concentration/nominal concentration) × 100.

IS* = Internal Standard.

Note. All QC samples were analyzed in triplicate. Mean values are reported.

Obs. = observed; Dev. = deviation.