Determination of urinary 6-mercaptopurine and three of its metabolites by HPLC-UV coupled with the iodine-azide reaction.

Abstract

BACKGROUND:

The presented method is able to determine 6-mercaptopurine (6-MP), 6-thioguanine, 6-mercaptopurine riboside and 6-thioguanine riboside in urine, and is thereby dedicated to control of thiopurine therapy of children with acute lymphoblastic leukemia. Good separation of the mentioned compounds was achieved on a C18 stationary phase with a sodium azide and sodium heptane sulfonate solution, acetonitrile and water at ratio of 50:1:49 (v/v/v).

RESULTS:

Coefficient of regression is >0.99 for all linearity ranges. LOD and LOQ are 0.3, 0.4, 0.3, 0.8 and 0.4, 0.6, 0.5 and 0.9 nmol/ml of urine for 6-MP, 6-thioguanine, 6-mercaptopurine riboside and 6-thioguanine riboside, respectively. Intra- and inter-day recovery and RSD are close to 100% and less than 10%, respectively, for all investigated thiopurines.

CONCLUSION:

The elaborated method was successfully applied for detection and quantitation of 6-MP and its selected metabolites in patients' urine samples.

• Authors: Robert Zakrzewski, Kamila Borowczyk, Adam Łuczak, Wojciech Młynarski, Joanna Trelińska
• Journal: Bioanalysis (Bioanalysis) Vol. 5 Issue 8 Pg. 869-77 (Apr 2013) ISSN: 1757-6199 [Electronic] England
• PMID: 23590470 (Publication Type: Journal Article)

Chemical References

• Azides
• Iodine
• Mercaptopurine

Topics

• Azides (chemistry)
• Child
• Chromatography, High Pressure Liquid (methods)
• Humans
• Iodine (chemistry)
• Limit of Detection
• Mercaptopurine (metabolism, urine)
• Spectrophotometry, Ultraviolet (methods)
• Time Factors
• Urinalysis (methods)