Development and validation of a liquid chromatography-tandem mass spectrometry method for quantifying delamanid and its metabolite in small hair samples

J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Apr 15:1169:122467. doi: 10.1016/j.jchromb.2020.122467. Epub 2020 Dec 10.

Abstract

New all-oral regimens for rifampin-resistant tuberculosis (RR-TB) are being scaled up globally. Measurement of drug concentrations in hair assesses long-term drug exposure. Delamanid (DLM) is likely to be a key component of future RR-TB treatment regimens, but a method to describe its quantification in hair via liquid chromatography-tandem mass spectrometry (LC-MS/MS) has not previously been described. We developed and validated a simple, fast, sensitive, and accurate LC-MS/MS method for quantifying DLM and its metabolite DM-6705 in small hair samples. We pulverized and extracted two milligrams of hair in methanol at 37 °C for two hours, and diluted 1:1 with water. A gradient elution method eluted DLM, DM-6705, and the internal standard OPC 14714 within 3 min, bringing overall analysis time to 5.5 min. The method has limits of detection (LOD) of 0.0003 ng/mg for DLM and 0.003 ng/mg for DM-6705. The established linear dynamic ranges are 0.003-2.1 ng/mg and 0.03-21 ng/mg for DLM and DM-6705, respectively. Eleven of 12 participant hair samples had concentrations within DLM's linear dynamic range, while all 12 samples had concentrations within the quantifiable range for DM-6705. The ranges of concentrations observed in these clinical samples for DLM and DM-6705 were 0.004-0.264 ng/mg hair and 0.412-12.041 ng/mg hair respectively. We demonstrate that while DLM was detected in hair at very low levels, its primary metabolite DM-6705 had levels approximately 100 times higher. Measuring DM-6705 in hair may accurately reflect long-term adherence to DLM-containing regimens for drug-resistant TB.

Keywords: DM-6705; Delamanid; Drug-resistant tuberculosis; Hair analysis; LC-MS/MS; Objective adherence metrics.

MeSH terms

  • Chromatography, Liquid / methods*
  • Hair / chemistry*
  • Humans
  • Limit of Detection
  • Linear Models
  • Nitroimidazoles / analysis*
  • Nitroimidazoles / therapeutic use
  • Oxazoles / analysis*
  • Oxazoles / therapeutic use
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*
  • Tuberculosis
  • Tuberculosis, Multidrug-Resistant / drug therapy

Substances

  • Nitroimidazoles
  • OPC-67683
  • Oxazoles