Analysis of L-Asparaginase which is used in the treatment of acute lemfoblastic leukemia with chromatographic methods

L-asparaginase (L-ASPZ) is chemotherapeutic used since 1961 in treatment of acute lymphoblastic leukemia (ALL). It is an enzyme used as medicine in treatment of ALL. It performs its effect in ALL by reducing level of L-Asparagine (L-Asn) in plasma. In thesis, aspargine(Asn), glutamine (Gln), aspartic acid (Asp), glutamic acid (Glu) levels as well as the activity of L-asparaginase in treatment were studied by chromatographic methods. Asparaginase degrades asparagine to aspartic acid and ammonia, depriving tumor cells of asparagine. Tumor cell loaded with L-Asparaginase lacks nutrients and can't proliferate. TDM becomes important for us to understand at what dose, type, and who is beneficial or harmful to L-asparaginase. Thus, both effective dose will be found and the shortening of treatment period will contribute. Reliability and sensitivity problems are encountered due to cross reactions in immunoassay methods and interventions in spectrophotometric methods. Determination amino acids in plasma for activity of L-ASPZ was developed by HPLC-MS/MS method. Method was developed with HILIC column and as a mobile phase gradient acetonitrile and ammonium acetate solution used at 0.4 mL/min flow. Linear range of amino acids was found to be 500-5000 ng/mL for Asn, Asp and Glu, and 5-50 μg/mL for Gln, respectively. LOD, LOQ were found as 100-500 ng/mL for Asn, Asp and Glu, and 1-5 μg/mL for Gln, respectively. Method has been validated and successfully applied to plasma analyzes. Method is selective, reproducible and can be used safely in plasma analysis.