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

Ankur Sharma, Elangovan Thavathiru, Doris Mangiaracina Benbrook, Sukyung Woo
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引用次数: 8

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, 2.1×150 mm) column coupled with a Waters XBridge™ C-18 (3.5 μm, 2.1×10 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.

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小鼠和人血浆中SHetA2的高效液相色谱定量分析方法的建立和验证:在药代动力学研究中的应用。
背景:SHetA2是一种用于癌症治疗和预防的口服抗癌药物。本研究的目的是建立和验证一种简单、经济、灵敏的HPLC-UV方法,用于生物样品中SHetA2的定量,并将该方法应用于药物的药代动力学研究。方法:以2,3 -二苯基喹啉为内标,冷冻乙腈为液-液沉淀萃取,制备小鼠和人血浆样品。通过Waters XBridge™BEH 130 C18 (3.5 μm, 2.1×150 mm)柱和Waters XBridge™C-18 (3.5 μm, 2.1×10 mm)保护柱分离SHetA2和内标,流动相为65% v/v乙腈:蒸馏水,流速为0.18 ml/min,等压模式。在检测波长为341 nm,柱温为25℃下洗脱。结果:该方法在5-1000 ng/ml范围内对血浆中SHetA2进行了验证,定量下限为5 ng/ml。该方法在人血浆(79.9 ~ 81.8%)和小鼠血浆(95.4 ~ 109.2%)中回收率高,无基质效应。日间和日间的准确度和精密度研究表明,该方法具有特异性、敏感性和可靠性。稳定性研究表明,SHetA2术后在自动进样器中稳定20小时,在-80°C的血浆中稳定6周。通过配制等分液并在-80°C保存,可以避免重复的冷冻和解冻。该方法成功地应用于荷瘤裸鼠经静脉和口服给药后SHetA2的血浆药代动力学研究。结论:一种新的定量小鼠和人血浆中SHetA2的方法已被验证,并可用于荷瘤小鼠体内SHetA2的药代动力学评价。该方法将用于临床研究中SHetA2的定量。
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