Department of Internal Medicine, Division of Nephrology, Clinical Pharmacology and Hypertension, Virginia Commonwealth University, Richmond, VA 23298, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Oct 15;878(28):2891-5. doi: 10.1016/j.jchromb.2010.07.022. Epub 2010 Aug 3.
A simple and sensitive high-performance liquid chromatographic (HPLC) method utilizing fluorescence detection was developed for the determination of the phosphodiesterase type 5 inhibitor tadalafil in mouse plasma. This method utilizes a simple sample preparation (protein precipitation) with high recovery of tadalafil (∼98%), which eliminates the need for an internal standard. For constituent separation, the method utilized a monolithic C(18) column and a flow rate of 1.0mL/min with a mobile phase gradient consisting of aqueous trifluoroacetic acid (0.1% TFA in deionized water pH 2.2, v/v) and acetonitrile. The method calibration was linear for tadalafil in mouse plasma from 100 to 2000ng/mL (r>0.999) with a detection limit of approximately 40ng/mL. Component fluorescence detection was achieved using an excitation wavelength of 275nm with monitoring of the emission wavelength at 335nm. The intra-day and inter-day precision (relative standard deviation, RSD) values for tadalafil in mouse plasma were less than 14%, and the accuracy (percent error) was within -14% of the nominal concentration. The method was utilized on mouse plasma samples from research evaluating the potential cardioprotective effects of tadalafil on mouse heart tissue exposed to doxorubicin, a chemotherapeutic drug with reported cardiotoxic effects.
建立了一种利用荧光检测的简单灵敏的高效液相色谱（HPLC）方法，用于测定小鼠血浆中的磷酸二酯酶 5 抑制剂他达拉非。该方法采用简单的样品制备（蛋白质沉淀），他达拉非的回收率高（约 98%），无需内标。对于成分分离，该方法使用整体 C18 柱和 1.0mL/min 的流速，流动相梯度由三氟乙酸水溶液（0.1%TFA 在去离子水 pH2.2，v/v）和乙腈组成。该方法在 100 至 2000ng/mL 范围内对小鼠血浆中的他达拉非进行线性校准（r>0.999），检测限约为 40ng/mL。通过使用 275nm 的激发波长和 335nm 的发射波长监测实现了组分荧光检测。小鼠血浆中他达拉非的日内和日间精密度（相对标准偏差，RSD）值小于 14%，准确度（百分比误差）在名义浓度的-14%范围内。该方法用于研究评估他达拉非对阿霉素（一种具有报道的心脏毒性作用的化疗药物）暴露的小鼠心脏组织的潜在心脏保护作用的小鼠血浆样品。
