Police Anitha, Shankar Vijay Kumar, Narasimha Murthy S
Department of Pharmaceutics and Drug Delivery, University of Mississippi, MS 38677, USA.
Department of Pharmaceutics and Drug Delivery, University of Mississippi, MS 38677, USA; Institute for Drug Delivery & Biomedical Research, Bangalore, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Feb 15;1076:44-53. doi: 10.1016/j.jchromb.2018.01.010. Epub 2018 Jan 7.
Vigabatrin is used as first line drug in treatment of infantile spasms for its potential benefit overweighing risk of causing permanent peripheral visual field defects and retinal damage. Chronic administration of vigabatrin in rats has demonstrated these ocular events are result of GABA accumulation and depletion of taurine levels in retinal tissues. In vigabatrin clinical studies taurine plasma level is considered as biomarker for studying structure and function of retina. The analytical method is essential to monitor taurine levels along with vigabatrin and GABA. A RP-HPLC method has been developed and validated for simultaneous estimation of vigabatrin, GABA and taurine using surrogate matrix. Analytes were extracted from human plasma, rat plasma, retina and brain by simple protein precipitation method and derivatized by naphthalene 2, 3‑dicarboxaldehyde to produce stable fluorescent active isoindole derivatives. The chromatographic analysis was performed on Zorbax Eclipse AAA column using gradient elution profile and eluent was monitored using fluorescence detector. A linear plot of calibration curve was observed in concentration range of 64.6 to 6458, 51.5 to 5150 and 62.5 to 6258 ng/mL for vigabatrin, GABA and taurine, respectively with r ≥ 0.997 for all analytes. The method was successfully applied for estimating levels of vigabatrin and its modulator effect on GABA and taurine levels in rat plasma, brain and retinal tissue. This RP-HPLC method can be applied in clinical and preclinical studies to explore the effect of taurine deficiency and to investigate novel approaches for alleviating vigabatrin induced ocular toxicity.
氨己烯酸因其潜在益处大于导致永久性周边视野缺损和视网膜损伤的风险,被用作治疗婴儿痉挛症的一线药物。在大鼠中慢性给予氨己烯酸已证明这些眼部事件是视网膜组织中GABA积累和牛磺酸水平降低的结果。在氨己烯酸临床研究中,血浆牛磺酸水平被视为研究视网膜结构和功能的生物标志物。该分析方法对于监测氨己烯酸、GABA和牛磺酸的水平至关重要。已开发并验证了一种反相高效液相色谱(RP-HPLC)方法,用于使用替代基质同时测定氨己烯酸、GABA和牛磺酸。通过简单的蛋白质沉淀法从人血浆、大鼠血浆、视网膜和脑组织中提取分析物,并用萘-2,3-二甲醛进行衍生化,以产生稳定的荧光活性异吲哚衍生物。色谱分析在Zorbax Eclipse AAA柱上进行,采用梯度洗脱曲线,并用荧光检测器监测洗脱液。分别在氨己烯酸、GABA和牛磺酸浓度范围为64.6至6458、51.5至5150和62.5至6258 ng/mL时观察到校准曲线的线性图,所有分析物的r≥0.997。该方法已成功应用于估计大鼠血浆、脑和视网膜组织中氨己烯酸的水平及其对GABA和牛磺酸水平的调节作用。这种RP-HPLC方法可应用于临床和临床前研究,以探索牛磺酸缺乏的影响,并研究减轻氨己烯酸诱导的眼部毒性的新方法。