The introduced research presents a novel in vivo quantitative method for assay of mixtures of pregabalin and tramadol as a common combinations approved for treatment of neuropathic pain. Green analytical chemistry is a recently emerging science concerned with control of the use of chemicals harmful to the environment in various analytical methods. Consequently, a green high‐performance thin layer chromatography (HPTLC) method was achieved for determination of the mixture in human plasma and urine satisfying both analytical and environmental standards. The separation was achieved on HPTLC sheets using a separating mixture of ethanol–ethyl acetate–acetone–ammonia solution (8:2:1:0.05, by volume) as a mobile phase. The sheets were dried in air then scanned at two wavelengths. For tramadol, 220 nm was chosen; however, pregabalin is an unconjugated drug, so its determination was a challenge. Hence for pregabalin, the plates were sprayed with ethanolic solution of ninhydrin (3%, w/v), to obtain a conjugated complex, which could be assessed at 550 nm. Furthermore, the developed method fulfilled the US Food and Drug Administration validation guidelines, and proved to be useful in therapeutic drug monitoring of this combination. The Eco‐scale assessment protocol was implemented to determine the greenness profile of the applied method.

中文翻译：

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经生态评估和FDA验证的HPTLC方法测定人体生物液中的普瑞巴林和曲马多

引入的研究提出了一种新颖的体内普瑞巴林和曲马多混合物的定量分析方法，已批准用于治疗神经性疼痛。绿色分析化学是一门新兴的科学，涉及以各种分析方法控制对环境有害的化学物质的使用。因此，实现了一种绿色高效薄层色谱（HPTLC）方法，用于测定满足分析和环境标准的人血浆和尿液中的混合物。使用乙醇-乙酸乙酯-丙酮-氨溶液的分离混合物（体积比为8：2：1：0.05）作为流动相，可以在HPTLC薄板上实现分离。片材在空气中干燥，然后在两个波长下扫描。对于曲马多，选择了220 nm。然而，普瑞巴林是一种非结合药物，因此其测定是一个挑战。因此，对于普瑞巴林，将板用茚三酮的乙醇溶液（3％，w / v）喷雾，以获得缀合的复合物，其可以在550nm处评估。此外，所开发的方法满足了美国食品药品监督管理局的验证准则，并被证明可用于该组合的治疗性药物监测。实施了生态规模评估协议，以确定所应用方法的绿色特征。