Development and application of a high-throughput liquid chromatography–tandem mass spectrometric method for the simultaneous determination of thymosin α1 and its recombinant human form in plasma and urine

Publication date: 5 June 2019Source: Journal of Pharmaceutical and Biomedical Analysis, Volume 170Author(s): Tingting Wang, Lei Yin, Hao Wang, John Paul Fawcett, Jingkai GuAbstractThymosin α1 (Thymalfasin, Tα1) is a naturally occurring polypeptide widely used as an immune system enhancer for the treatment of HIV/AIDS, hepatitis B and C, and cancer. Recombinant human Tα1 (rh-Tα1) lacking N-terminal acetylation (NTA) displays similar biological activity to Tα1 and has completed phase III clinical trials in China. To compare the pharmacokinetics of rh-Tα1 and Tα1 and establish whether they undergo mutual transmutation in vivo, we developed a novel bioassay based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) for the simultaneous determination of the two peptides in human plasma and urine. Sample preparation by protein precipitation using a mixture of methanol and perchloric acid was followed by HPLC on a Zorbax 300SB-C18 column (150 × 4.6 mm, 5 μm) maintained at 40 °C. Detection was by multiple reaction monitoring (MRM) of the precursor-to-product ion transitions at m/z 778.0→316.0 for Tα1, m/z 767.3→955.0 for rh-Tα1 and m/z 832.3→159.2 for the internal standard, eptifibatide. The method was linear in the range 2–100 ng/mL for both analytes with good accuracy and precision. High sample throughput was facilitated by inclusion of a parallel two-column chromatographic system. The method was successfully applied to a comparative phar...
Source: Journal of Pharmaceutical and Biomedical Analysis - Category: Drugs & Pharmacology Source Type: research