LC-MS/MS method development, validation, and application for levonorgestrel assay from contraceptive implants in Bangladeshi women
- 1Department of Pharmaceutical Technology, University of Dhaka
- 2BCSIR Dhaka Laboratories, Bangladesh Council of Scientific and Industrial Research (BCSIR)
- 3Department of Pharmaceutical Technology, Jadavpur University
Abstract
Levonorgestrel (LNG) used in contraceptive methods, necessitates accurate pharmacokinetic (PK) profiling to ensure efficacy and safety. The objective of this study was to develop and validate a highly sensitive LC-MS/MS method for the quantification of LNG in human serum and its application in a pharmacokinetic study. 500 µL sample was processed using a liquid-liquid extraction (LLE) protocol optimized for LNG recovery that yielded consistent analyte recovery while minimizing matrix interferences. Chromatographic separation was performed on Waters ACQUITY UPLC BEH C18 reverse-phase analytical column (2.1 × 100 mm, 1.7 μm) using mobile phase consisting of acetonitrile and water with 0.1% formic acid (70:30, v/v), delivered under an isocratic elution at 300 µL/min flow rate. These conditions provided sharp peak resolution and minimized run times. Mass spectrometric analysis was performed with Waters XEVO TQ ABSOLUTE triple quadrupole instrument operating in positive electrospray ionization mode and monitored LNG using multiple reaction monitoring with precursor ion at m/z 312.98 > 81.468, 90.921, 108.963 amu. Optimized fragmentation parameters yielded high signal-to-noise ratios, enabling trace-level detection in biological matrices. The method demonstrated linearity over calibration ranges of 32.5–2000 pg/mL, with co-efficient of determination (R²) exceeding 0.997. Precision and accuracy met bioanalytical method validation guidelines, confirming the robustness of the assay for PKs. The validated method was successfully employed to measure serum LNG level in a PK study among women implanted in Bangladesh. The method’s sensitivity facilitated LNG detection supporting the assessment of drug-release performance in clinical settings. The method proved highly reliable, sensitive, and specific for quantifying LNG in human serum, supporting its use in both clinical monitoring and research applications.
Related
- Development, Validation, and Application of a Highly Sensitive RP-HPLC Method for the Determination and Release Profile Comparison of Levonorgestrel from Silicone-Based Contraceptive Implants
- Validation of a MALDI Intact Cell Mass Spectrometry Workflow for Routine Application in Monitoring Industrial E. coli Fermentations
- 本科生综合实验：基于卤键有机框架材料的分散固相萃取方法构建及在溴代阻燃剂分析中的应用 A comprehensive undergraduate experiment： development of a dispersive solid-phase extraction method based on halogen-bonded organic frameworks and its application to the analysis of brominated flame retardants
- Development of a method for the preparation of zirconium-89 radiolabelled chitosan nanoparticles as an application for leukocyte trafficking with positron emission tomography
- HPTLC Method Development and Validation for Simultaneous Estimation of Rifaximin and Metronidazole Benzoate in Combined Tablet Dosage Form
BibTeX
@article{kabir2026lcmsms,
title = {{LC-MS/MS method development, validation, and application for levonorgestrel assay from contraceptive implants in Bangladeshi women}},
author = {Abul Kalam Lutful Kabir and Md. Najem Uddin and Satyajit Roy Rony and Sitesh Chandra Bachar and Emdad Hossain},
journal = {Scientific Reports},
year = {2026},
month = aug,
doi = {10.1038/s41598-026-63325-1},
url = {https://doi.org/10.1038/s41598-026-63325-1},
note = {CC BY},
}
RIS
TY - JOUR
TI - LC-MS/MS method development, validation, and application for levonorgestrel assay from contraceptive implants in Bangladeshi women
AU - Abul Kalam Lutful Kabir
AU - Md. Najem Uddin
AU - Satyajit Roy Rony
AU - Sitesh Chandra Bachar
AU - Emdad Hossain
PY - 2026
DA - 2026/08/01
JO - Scientific Reports
DO - 10.1038/s41598-026-63325-1
UR - https://doi.org/10.1038/s41598-026-63325-1
AB - Levonorgestrel (LNG) used in contraceptive methods, necessitates accurate pharmacokinetic (PK) profiling to ensure efficacy and safety. The objective of this study was to develop and validate a highly sensitive LC-MS/MS method for the quantification of LNG in human serum and its application in a pharmacokinetic study. 500 µL sample was processed using a liquid-liquid extraction (LLE) protocol optimized for LNG recovery that yielded consistent analyte recovery while minimizing matrix interferences. Chromatographic separation was performed on Waters ACQUITY UPLC BEH C18 reverse-phase analytical column (2.1 × 100 mm, 1.7 μm) using mobile phase consisting of acetonitrile and water with 0.1% formic acid (70:30, v/v), delivered under an isocratic elution at 300 µL/min flow rate. These conditions provided sharp peak resolution and minimized run times. Mass spectrometric analysis was performed with Waters XEVO TQ ABSOLUTE triple quadrupole instrument operating in positive electrospray ionization mode and monitored LNG using multiple reaction monitoring with precursor ion at m/z 312.98 > 81.468, 90.921, 108.963 amu. Optimized fragmentation parameters yielded high signal-to-noise ratios, enabling trace-level detection in biological matrices. The method demonstrated linearity over calibration ranges of 32.5–2000 pg/mL, with co-efficient of determination (R²) exceeding 0.997. Precision and accuracy met bioanalytical method validation guidelines, confirming the robustness of the assay for PKs. The validated method was successfully employed to measure serum LNG level in a PK study among women implanted in Bangladesh. The method’s sensitivity facilitated LNG detection supporting the assessment of drug-release performance in clinical settings. The method proved highly reliable, sensitive, and specific for quantifying LNG in human serum, supporting its use in both clinical monitoring and research applications.
SN - 2045-2322
ER -