Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, Pamplona, Spain.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Nov 15;879(30):3490-6. doi: 10.1016/j.jchromb.2011.09.029. Epub 2011 Sep 22.
An ultra high performance liquid chromatography tandem mass spectrometry method (UHPLC-MS/MS) was developed and validated for the quantitation of LBH589, a novel histone deacetylase inhibitor (HDACi), in mouse plasma and tissues (liver, spleen, kidney and lung). Tobramycin was employed as the internal standard. Separation was performed on an Acquity UPLC™ BEH column, with a mobile phase consisting of 10% water (with 0.1% of trifluoroacetic acid) and 90% methanol (with 0.1% trifluoroacetic acid). LBH589 and tobramycin were determined using an electrospray ionization (ESI) interface. Detection was performed on electrospray positive ionization mass spectrometry by multiple reaction monitoring of the transitions of LBH589 at m/z 349.42→157.95 and of tobramycin at 468.2→163. Calibration curves for the UHPLC method (0.0025-1 μg/mL for plasma and tissue homogenates, equivalent to 0.0357-14.2857 μg/g for tissue samples) showed a linear range of detector responses (r>0.998). Intra-batch and inter-batch precision expressed as coefficient of variation (CV) ranged from 0.92 to 8.40%. Accuracy expressed as bias, ranged from -2.41 to 2.62%. The lower limit of quantitation (LLOQ) was 0.0025 μg/mL for both plasma and tissue homogenate samples, equivalent to 0.0357 μg/g tissue. This method was successfully applied to quantify LBH589 in plasma and tissue samples obtained after the intraperitoneal administration of a single dose of 20 mg/kg of LBH589 in BALB/c mice.
建立并验证了一种超高效液相色谱-串联质谱法（UHPLC-MS/MS），用于定量检测新型组蛋白去乙酰化酶抑制剂（HDACi）LBH589 及其在小鼠血浆和组织（肝、脾、肾和肺）中的浓度。妥布霉素被用作内标。分离在 Acquity UPLC™ BEH 柱上进行，流动相由 10%水（含 0.1%三氟乙酸）和 90%甲醇（含 0.1%三氟乙酸）组成。LBH589 和妥布霉素采用电喷雾电离（ESI）接口进行检测。通过监测 LBH589 在 m/z 349.42→157.95 和妥布霉素在 468.2→163.的转化，采用正离子电喷雾质谱进行多反应监测，对 LBH589 进行检测。UHPLC 方法的校准曲线（血浆和组织匀浆为 0.0025-1μg/mL，相当于组织样品为 0.0357-14.2857μg/g）显示出检测响应的线性范围（r>0.998）。批内和批间精密度以变异系数（CV）表示，范围为 0.92-8.40%。以偏倚表示的准确度，范围为-2.41-2.62%。定量下限（LLOQ）对于血浆和组织匀浆样品均为 0.0025μg/mL，相当于组织样品为 0.0357μg/g。该方法成功应用于检测 BALB/c 小鼠单次腹腔注射 20mg/kg LBH589 后血浆和组织样品中的 LBH589。
