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

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.


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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