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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >A rapid, ideal, and eco-friendlier protocol for quantifying proline
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A rapid, ideal, and eco-friendlier protocol for quantifying proline

机译：一种快速，理想且生态友好的方案，可定量脯氨酸

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Proline, a stress marker, is routinely quantified by a protocol that essentially uses hazardous toluene. Negative impacts of toluene on human health prompted us to develop a reliable alternate protocol for proline quantification. Absorbance of the proline-ninhydrin condensation product formed by reaction of proline with ninhydrin at 100 A degrees C in the reaction mixture was significantly higher than that recorded after its transfer to toluene, revealing that toluene lowers sensitivity of this assay. lambda (max) of the proline-ninhydrin complex in the reaction mixture and toluene were 508 and 513 nm, respectively. Ninhydrin in glacial acetic acid yielded higher quantity of the proline-ninhydrin condensation product compared to ninhydrin in mixture of glacial acetic acid and H3PO4, indicating negative impact of H3PO4 on proline quantification. Further, maximum yield of the proline-ninhydrin complex with ninhydrin in glacial acetic acid and ninhydrin in mixture of glacial acetic acid and H3PO4 was achieved within 30 and 60 min, respectively. This revealed that H3PO4 has negative impact on the reaction rate and quantity of the proline-ninhydrin complex formed. In brief, our proline quantification protocol involves reaction of a 1-ml proline sample with 2 ml of 1.25 % ninhydrin in glacial acetic acid at 100 A degrees C for 30 min, followed by recording absorbance of the proline-ninhydrin condensation product in the reaction mixture itself at 508 nm. Amongst proline quantification protocols known till date, our protocol is the most simple, rapid, reliable, cost-effective, and eco-friendlier.
机译：脯氨酸是一种压力标记物，通常通过基本使用有害甲苯的方案进行定量。甲苯对人类健康的负面影响促使我们开发出可靠的替代方案进行脯氨酸定量。在100 A的温度下，脯氨酸与茚三酮反应形成的脯氨酸-茚三酮缩合产物在反应混合物中的吸光度明显高于转移至甲苯后记录的吸光度，表明甲苯降低了该测定的灵敏度。反应混合物和甲苯中脯氨酸-茚三酮配合物的λ（max）分别为508和513 nm。与冰醋酸和H3PO4混合物中的茚三酮相比，冰醋酸中的茚三酮生成的脯氨酸-茚三酮缩合产物量更高，这表明H3PO4对脯氨酸定量有负面影响。此外，分别在30分钟和60分钟内获得了脯氨酸-茚三酮与冰醋酸中的茚三酮和冰醋酸和H3PO4的混合物中的茚三酮的最大产率。这表明H 3 PO 4对所形成的脯氨酸-茚三酮配合物的反应速率和数量具有负面影响。简而言之，我们的脯氨酸定量方案包括将1 ml脯氨酸样品与2 ml 1.25％茚三酮在冰醋酸中在100 A的温度下反应30分钟，然后记录反应中脯氨酸-茚三酮缩合产物的吸光度混合物本身在508 nm。在迄今为止已知的脯氨酸定量方案中，我们的方案是最简单，快速，可靠，具有成本效益和生态友好的。

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