Development and Validation of an HPLC Analytical Method to Determine 6-Merpactopurine Concentration in Oral Suspension

Title:Development and Validation of an HPLC Analytical Method to Determine 6-Merpactopurine Concentration in Oral Suspension

Volume: 9

Author(s): Kerley Cristiane Victorino Romão, Fernando Luiz Affonso Fonseca, Fernanda Schindler, Marina Cristina Peres, Glaucia Luciano da Veiga, Edimar Cristiano Pereira and Beatriz da Costa Aguiar Alves*

Affiliation:

• Laboratório de Análises, Clínicas do Centro Universitário, Faculdade de Medicina do ABC, Av. Príncipe de Gales, 821, 09060-650, Santo André, SP, Brazil

Keywords: 6-Mercaptopurine, oral suspension, HPLC, quantitative analysis, pediatric oncology, oral drug.

Abstract:

Background: The 6-mercaptopurine is an active ingredient used to treat certain types of leukemia. This drug is an immunosuppressive and antineoplastic agent that belongs to the thiopurine class. In Brazil, 6-MP is currently available only in the form of 50 mg tablets, sold as Purinethol® and manufactured by Glaxo Smith Kline. The lack of the liquid formulation’s production impedes treatment, assuming that one of its advantages is through its applicability in pediatric patients, who show the highest incidence among others.

Objective: The purpose of this work was to evaluate the development and application of a reversed phase high performance liquid chromatography (HPLC) method using an Agilent 1220 Infinity® G4294B chromatograph with photodiode array detector.

Methods: HPLC assays were performed on an Eclipse plus® C18 column (4.6 x 150 mm, 3.5 μm particle size) using a gradient mode mixture of acetonitrile and aqueous acetic acid solution as a mobile phase, with a flow of 1 mL.min-1 and detection at 324 nm. The method was validated by determining its selectivity, linearity, precision, accuracy, and robustness.

Results: Retention time for 6-mercaptopurine was 5.12 minutes. The detector’s response was linear at concentrations from 1.6 to 2.4 μg/mL. The results of the method’s accuracy evaluation showed mean recovery of the amount of substance added to the samples of between 99.88 and 100.5%. For precision, repeatability and intermediate precision were evaluated. The repeatability showed a standard deviation of 0.0737. The intermediate precision was assessed on three different. For three days of the studies, the values of the standard deviations were less than 3%, showing repeatability and intermediate precision adequate for the analytical method in question. The limit of detection was determined as 3.6 ng/mL. The limit of quantification was determined as 12 ng/mL. The chromatographic method was robust.

Conclusion: The proposed method can be applied to control the quality of 6-MP oral suspension to ensure that the required content is delivered to pediatric oncology patients.

Cite this article as:

Romão Cristiane Victorino Kerley, Fonseca Luiz Affonso Fernando, Schindler Fernanda, Peres Cristina Marina, da Veiga Luciano Glaucia, Pereira Cristiano Edimar and da Costa Aguiar Alves Beatriz*, Development and Validation of an HPLC Analytical Method to Determine 6-Merpactopurine Concentration in Oral Suspension, Current Chromatography 2022; 9 (1) : e260122200506 . https://dx.doi.org/10.2174/2213240609666220126101414