Transl Clin Pharmacol.  2018 Sep;26(3):134-140. 10.12793/tcp.2018.26.3.134.

Development of a UPLC-MS/MS method for the therapeutic monitoring of L-asparaginase

• Jeong HC
• Kim T
• Yang DH
• Shin KH

• KMID: 2420296
• DOI: http://doi.org/10.12793/tcp.2018.26.3.134

Abstract

This study aimed to develop a UPLC-MS/MS method for determining plasma levels of L-aspartic acid and L-asparagine and the activity of L-asparaginase. L-aspartic acid, L-asparagine, and L-aspartic acid-2,3,3-d3 were extracted from human plasma by protein precipitation with sulfosalicylic acid (30%, v/v). The plasma samples were analyzed using an Imtakt Intrada amino acid analysis column with 25 mM ammonium formate and 0.5% formic acid in acetonitrile as the mobile phase with step gradient method at a flow rate of 0.5 mL/min. The injection volume was 5 µL, and the total run time was 15 min. Inter- and intra-batch accuracies (%) ranged from 96.62-106.0% for L-aspartic acid and 89.85-104.8%, for L-asparagine, and the coefficient of variation (CV%) did not exceed 7%. The validation results for L-aspartic acid and L-asparagine satisfied the specified criterion, however, the results for L-asparaginase activity assay showed a borderline validity. This study could be a foundation for further development of therapeutic drug monitoring systems using UPLC-MS/MS.

Keyword

L-asparaginase; L-asparagine; L-aspartic acid; UPLC-MS/MS

MeSH Terms

Ammonium Compounds
Asparagine
Aspartic Acid
Drug Monitoring
Humans
Methods*
Plasma
Asparagine
Aspartic Acid

Figure

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  Figure 1 Product ion spectra and fragmentation patterns of (A) L-aspartic acid, (B) L-asparagine, and (C) L-aspartic acid-2,3,3-d3 (IS) by collision-induced dissociation of the corresponding parention.

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  Figure 2 The chromatogram of (A) L-aspartic acid and (B) L-asparagine in lower limit of quantification (LLOQ) sample (LLOQ = 1.95 µM).

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  Figure 3 The standard curve of L-asparaginase activity assay.

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