Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, C/Irunlarrea 1, E-31008 Pamplona, Spain.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Sep 15;935:47-53. doi: 10.1016/j.jchromb.2013.07.020. Epub 2013 Jul 29.
A simple liquid-liquid extraction procedure and quantification by high-performance liquid chromatography (HPLC) method coupled to a diode-array detector (DAD) of genistein (GEN) was developed in various mouse biological matrices. 7-ethoxycoumarin was used as internal standard (IS) and peaks were optimally separated using a Kinetex C18 column (2.6μm, 150mm×2.10mm I.D.) at 40°C with an isocratic elution of mobile phase with sodium dihydrogen phosphate 0.01M in water at pH 2.5 and methanol (55:45, v/v), at a flow rate of 0.25mL/min. The injection volume was 10μL. In all cases, the range of GEN recovery was higher than 61%. The low limit of quantification (LLOQ) was 25ng/mL. The linearity of the calibration curves was satisfactory in all cases as shown by correlation coefficients >0.996. The within-day and between-day precisions were <15% and the accuracy ranged in all cases between 90.14% and 106.05%. This method was successfully applied to quantify GEN in liver, spleen, kidney and plasma after intravenous administration of a single dose (30mg/kg) in female BALB/C mice.
建立了一种在各种小鼠生物基质中通过高效液相色谱（HPLC）法结合二极管阵列检测器（DAD）测定染料木黄酮（GEN）的简单液-液萃取程序和定量方法。7-乙氧基香豆素被用作内标（IS），使用 Kinetex C18 柱（2.6μm，150mm×2.10mm ID）在 40°C 下，以水的 0.01M 磷酸二氢钠和甲醇（55:45，v/v）为流动相进行等度洗脱，流速为 0.25mL/min。进样体积为 10μL。在所有情况下，GEN 的回收率均高于 61%。最低定量限（LLOQ）为 25ng/mL。所有情况下校准曲线的线性均令人满意，相关系数均大于 0.996。日内和日间精密度均<15%，准确度均在 90.14%至 106.05%之间。该方法成功应用于雌性 BALB/C 小鼠单次静脉注射（30mg/kg）后测定肝脏、脾脏、肾脏和血浆中的 GEN。
