Moussata Justin, Wang Zhijun, Wang Jeffrey
Department of Pharmaceutical Sciences, College of Pharmacy, Western University of Health Sciences, 309 E. 2(nd) Street, Pomona, CA 91766, USA.
Center for Advancement of Drug Research and Evaluation, College of Pharmacy, Western University of Health Sciences, 309 E. 2(nd) Street, Pomona, CA 91766, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 May 1;958:1-9. doi: 10.1016/j.jchromb.2014.02.026. Epub 2014 Mar 12.
A novel Indole-3-carbinol derivative (I3C) prodrug, indole-3-carbinol acetate (I3CA), was synthesized and a rapid high-performance liquid chromatography (HPLC) method for the quantification of I3CA, I3C, and the major metabolite of I3C, diindolylmethane (DIM), in mouse plasma, liver and kidney tissues was developed and validated. 4-Methoxy-1-methylindole was used as the internal standard. Chromatographic separation was achieved on a Symmetry(®) C18 column (75mm×4.6mm, 3.5μm) and the detection was made at 280nm. A gradient elution was programmed with the mobile phases of water (A) and acetonitrile (B) and a flow rate of 1ml/min. The total run time was 15min. The calibration curves were linear over the range of 0.06-1.6μg/ml for both I3C and DIM with a correlation coefficient (r(2)) higher than 0.997 and the lower limit of quantitation (LLOQ) of 0.06μg/ml. The calibration curve of I3CA was linear over the range of 0.15-4.0μg/ml, with a r(2)>0.995 and LLOQ of 0.15μg/ml. I3CA, I3C, and DIM intra-day accuracy values of plasma, liver and kidney samples ranged from 90.0 to 101.3%, while the inter-day ones were between 93.3 and 101.9%. Precision evaluated by the relative standard deviation was ranged from 2.0 to 14.8% for intra-day and 1.9 to 14.4% for inter-day variability. I3CA, I3C, and DIM were stable in mouse plasma, liver and kidney samples containing an esterase inhibitor dichlorvos. This method was successfully applied to a pharmacokinetic study in mice following oral and intravenous administration of I3C and I3CA.
合成了一种新型吲哚 - 3 - 甲醇衍生物（I3C）前药——吲哚 - 3 - 甲醇乙酸酯（I3CA），并建立和验证了一种快速高效液相色谱（HPLC）方法，用于定量小鼠血浆、肝脏和肾脏组织中的I3CA、I3C以及I3C的主要代谢产物二吲哚甲烷（DIM）。以4 - 甲氧基 - 1 - 甲基吲哚作为内标。在Symmetry(®) C18柱（75mm×4.6mm，3.5μm）上实现色谱分离，检测波长为280nm。流动相为水（A）和乙腈（B），采用梯度洗脱，流速为1ml/min。总运行时间为15分钟。I3C和DIM的校准曲线在0.06 - 1.6μg/ml范围内呈线性，相关系数（r(2)）高于0.997，定量下限（LLOQ）为0.06μg/ml。I3CA的校准曲线在0.15 - 4.0μg/ml范围内呈线性，r(2)>0.995，LLOQ为0.15μg/ml。血浆、肝脏和肾脏样品中I3CA、I3C和DIM的日内准确度值在90.0%至101.3%之间，而日间准确度值在93.3%至101.9%之间。通过相对标准偏差评估的精密度，日内变异性范围为2.0%至14.8%，日间变异性范围为1.9%至14.4%。I3CA、I3C和DIM在含有酯酶抑制剂敌敌畏的小鼠血浆、肝脏和肾脏样品中稳定。该方法成功应用于小鼠口服和静脉注射I3C和I3CA后的药代动力学研究。
