Sharma Ankur, Thavathiru Elangovan, Benbrook Doris Mangiaracina, Woo Sukyung
Department of Pharmaceutical Sciences, College of Pharmacy, University of Oklahoma Health Sciences Center, 1110 N. Stonewall Ave. CPB331, Oklahoma City, Oklahoma 73117-1200, USA.
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.
J Pharm Technol Drug Res. 2017;6. doi: 10.7243/2050-120X-6-2.
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.
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.
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.
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.
SHetA2是一种正在进行癌症治疗与预防研究的口服抗癌药物。本研究的目的是开发并验证一种简单、经济高效且灵敏的HPLC-UV方法，用于定量生物样品中的SHetA2，并将该方法应用于该药物的药代动力学研究。
小鼠和人血浆的样品制备包括液-液沉淀，并用冷乙腈以2,3-二苯基喹喔啉作为内标进行萃取。SHetA2和内标的分离通过Waters XBridge™ BEH 130 C18（3.5μm，2.1×150mm）色谱柱与Waters XBridge™ C-18（3.5μm，2.1×10mm）保护柱联用实现，使用65% v/v乙腈：蒸馏水作为流动相，等度洗脱模式，流速为0.18ml/min。分析物在341nm检测波长、25°C柱温下洗脱。
该方法在血浆中SHetA2浓度范围为5-1000ng/ml时得到验证，定量下限为5ng/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。日内和日间准确度与精密度研究表明该方法特异、灵敏且可靠。稳定性研究表明，SHetA2在自动进样器中术后20小时稳定，在血浆中-80°C下可稳定六周。通过制备等分试样并储存在-80°C可避免反复冻融。所开发的方法成功应用于研究荷瘤裸鼠静脉注射和口服给药后SHetA2的血浆药代动力学。
一种用于定量小鼠和人血浆中SHetA2的新方法已得到验证，并正在用于荷瘤小鼠中SHetA2的药代动力学评估。所开发的方法将用于临床研究中SHetA2的定量。
