Quantifying plasma dacarbazine levels in advanced melanoma patients: a liquid chromatography-tandem mass spectrometry performance analysis

Abstract

The aim of this study was to develop a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying dacarbazine levels in the plasma of advanced melanoma patients, followed by an assessment of its analytical capabilities. The research encompassed the design of a high-performance liquid chromatography (HPLC) system, with the quantitative analysis performed using the multiple reaction monitoring (MRM) techniques and specific ion transition: 181.0 > 152.5 for dacarbazine and 187.1 > 158.6 for the internal standard (IS), dacarbazine-D6. The validation of the method involved an evaluation of parameters including linearity, detection limit, precision, and accuracy. Notably, the linear range extended from 10 to 1,000 µg/L for dacarbazine, and the method exhibited a detection limit of 10 µg/L. The method’s precision, indicated by within-run and between-run coefficients of variation (CV), both being ≤4.2% and ≤8.3%, respectively. Furthermore, the accuracy of measurements, ranging from 86.1% to 99.4%, underscored the method’s reliability. In clinical application, the dacarbazine levels of healthy control (n = 20) were 0.6 ± 0.02 μg/L; 770.9 ± 203.2 μg/mL in early-stage-melanoma patients (n = 22), and 588.7 ± 153.2 μg/mL in advanced melanoma patients (n = 25). The results serve as clinical evidence showing that long-term dacarbazine treatment affects the metabolism of dacarbazine.

Keywords:

• LC-MS/ms
• mRM
• dacarbazine
• plasma level of dacarbazine and melanoma

Acknowledgments

Funding from Hematology Oncology Cancer Center GWV, 1000 E Mountain Blvd, Wilkes-Barre, PA 18711.

Author contributions

All the data generation, collection, and analysis were done by Wenhan Cheng

All the urine samples were collected from Geisinger Medical Center - Laboratory Medicine, Pennsylvania. The sample collection was done by Dr. Jackson (Geisinger Medical Center).

Ethical approval and IRB review statement

This clinical research and patient recruitment is following the ethics and regulations, which include: the Nuremberg Code (1947), Declaration of Helsinki (2000), Belmont Report (1979), CIOMS (2002), and U.S. Common Rule (1991). There are seven main principles that have been applied to this clinical research and patient recruitment.

Reviewed by Institutional Review Board

NAME: Geisinger Institutional Review Board

The study protocol was reviewed and approved by the Geisinger Clinic Human Research Protection Office (IRB REGISTRATION #: 00008345). All participants provided informed consent prior to specimen and data collection).

INSTITUTION: Geisinger Clinic

FWA ASSURANCE: FWA00000063

ACCREDITATION: Geisinger Health System is accredited by the Association for the Accreditation of Human Protection Programs, Inc. (AAHRPP)

ADDRESS: Geisinger Institutional Review Board, 100 North Academy Avenue. Danville, PA 17822-3069.

Informed consent

Informed consent was obtained from all individuals included in this study.

Disclosure statement

No potential conflict of interest was reported by the authors.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.