Journal of Pharmaceutical Technology & Drug Research

Journal of Pharmaceutical Technology &
Drug Research

ISSN 2050-120X
Original Research

Bioanalytical method development and validation of HPLCUV assay for the quantification of SHetA2 in mouse and human plasma: Application to pharmacokinetics study

Ankur Sharma1, Elangovan Thavathiru2, Doris Mangiaracina Benbrook1,2 and Sukyung Woo1*

*Correspondence: Sukyung Woo sukyung-woo@ouhsc.edu

1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Oklahoma Health Sciences Center, 1110 N. Stonewall Ave. CPB331, Oklahoma City, Oklahoma 73117-1200, USA.

Author Affiliations

2, Department of Obstetrics and Gynecology, Stephenson Cancer Center (SCC), University of Oklahoma Health Sciences Center, 975 NE 10th St, BRC 1217A, Oklahoma City, Oklahoma 73104, USA.

Abstract

Background: SHetA2 is an oral anticancer agent being investigated for cancer treatment and prevention. The aim of this study was to develop and validate a simple, cost-effective, and sensitive HPLC-UV method for the quantification of SHetA2 in biological samples and to apply the method to pharmacokinetic studies of the drug.

Methods: Sample preparation for mouse and human plasmas involved liquid-liquid precipitation and extraction using chilled acetonitrile with 2, 3-Diphenylquinoxaline as an internal standard. The separation of SHetA2 and internal standard was achieved via Waters XBridge™ BEH 130 C18 (3.5 μm, 2.1x150 mm) column coupled with a Waters XBridge™ C-18 (3.5 μm, 2.1x10 mm) guard column using 65% v/v acetonitrile: distilled water as a mobile phase in an isocratic mode with a flow rate of 0.18 ml/min. The analytes were eluted at a detection wavelength of 341 nm at a column temperature of 25°C.

Results: The method was validated across a range of 5-1000 ng/ml for SHetA2 in plasma, with a lower limit of quantification of 5 ng/ml. The method showed high recovery in human (79.9-81.8%) and mouse (95.4-109.2%) plasma with no matrix effect. The intra- and inter-day accuracy and precision studies demonstrated that the method was specific, sensitive, and reliable. Stability studies showed that SHetA2 is stable for 20 h postoperatively in the auto sampler, and for six weeks at -80°C in plasma. Repetitive freezing and thawing may be avoided by preparing the aliquots and storing them at -80°C. The developed method was successfully applied to study the plasma pharmacokinetics of SHetA2 in tumor-bearing nude mice after intravenous and oral administration.

Conclusion: A novel method for quantifying SHetA2 in mouse and human plasmas has been validated and is being applied for pharmacokinetic evaluation of SHetA2 in tumor-bearing mice. The developed method will be utilized for the quantification of SHetA2 in clinical studies.

Keywords: SHetA2, HPLC, Human plasma, Mouse plasma, Preclinical pharmacokinetics

ISSN 2050-120X
Volume 6
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