HPLC-MS/MS Method for Monitoring of L-Asparaginase Activity by Using Asparagine and Aspartic Acid Plasma Levels


Kaplan C., Daniş İ., Ünal D. Ş.

Journal of Mass Spectrometry, vol.61, no.3, 2026 (SCI-Expanded, Scopus) identifier identifier

  • Publication Type: Article / Article
  • Volume: 61 Issue: 3
  • Publication Date: 2026
  • Doi Number: 10.1002/jms.70036
  • Journal Name: Journal of Mass Spectrometry
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Chemical Abstracts Core, Chimica, Compendex, EMBASE, MEDLINE
  • Keywords: acute lymphoblastic leukemia, amino acid analysis, L-asparaginase activity, mass spectrometry, therapeutic drug monitoring
  • Istanbul University Affiliated: Yes

Abstract

The effectiveness of L-asparaginase and therefore the effectiveness of acute lymphoblastic leukemia treatment will be understood by determining the substrates of the enzyme, L-asparagine and L-glutamine. For this purpose, the high-performance liquid chromatography–tandem mass spectrometry method was developed and validated by analyzing the L-asparaginase substrates asparagine and glutamine and its products aspartic acid and glutamic acid from plasma. Acetonitrile and ammonium acetate were used at 0.4 mL/min in gradient mobile phase flow using a HILIC column for chromatographic separations. The linear amino acid range was found to be 500–5000 ng/mL for asparagine, aspartic acid, and glutamic acid, and 5–50 μg/mL for glutamine, respectively. Detection limit and quantitation limit were found to be 100–500 ng/mL for asparagine, aspartic acid, and glutamic acid, and 1–5 μg/mL for glutamine, respectively. The validated method has been successfully applied to plasma samples. The method was found to be selective and reproducible.