A three-phase hollow fiber liquid-phase microextraction based on reversed micellar supramolecule was developed and employed for the determination of quality markers of three anthraquinones in Rhubarb and their plasma protein binding rates. Moreover, the extraction mechanism of reversed micellar supramolecule and determination mechanism of plasma protein binding rate by the proposed method are both explored. In this procedure, n-nonanol was impregnated in the wall pores and reverse micelle of tetrabutylammonium chloride/n-nonanol filled in lumen of hollow fibers, which were covered with a thin salt film and used for the microextraction of target compounds prior to HPLC analysis. Important extraction parameters including extraction solvent type, reversed micellar supramolecule type and concentration, sample phase pH, salt concentration, stirring rate, extraction time, and binding time between active compound and plasma protein were investigated. Under the optimized conditions, limits of detection were 0.1–0.4 ng/mL with enrichment factors in the range of 54.2–100.7. The linearities were 0.5–500 ng/mL with r2 ≥ 0.9838. Satisfactory accuracies (relative recoveries 95.7–98.3%) were also obtained. The average plasma protein binding rates of rhein, chrysophenol, and physcion were 96.7%, 41.8%, and 46.7%, respectively. The results showed that the proposed procedure can not only extract and concentrate anthraquinones in Rhubarb, but also determine their plasma protein binding rates.

中文翻译：

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反胶束中空纤维液相微萃取联用高效液相色谱法测定大黄中蒽醌的 Q 标志物及其血浆蛋白结合率

开发了一种基于反胶束超分子的三相中空纤维液相微萃取方法，用于大黄中三种蒽醌类物质的质量标志物及其血浆蛋白结合率的测定。此外，还探讨了该方法对反胶束超分子的提取机理和血浆蛋白结合率的测定机制。在这个过程中，正壬醇浸渍在壁孔中，四丁基氯化铵/正壬醇的反胶束填充在空心纤维的管腔中，并覆盖有一层薄薄的盐膜，用于在 HPLC 分析之前对目标化合物进行微萃取. 重要的提取参数包括提取溶剂类型、反胶束超分子类型和浓度、样品相 pH 值、盐浓度、研究了搅拌速率、提取时间和活性化合物与血浆蛋白之间的结合时间。在优化条件下，检测限为 0.1-0.4 ng/mL，富集因子在 54.2-100.7 范围内。线性为 0.5–500 ng/mL，r2 ≥ 0.9838。还获得了令人满意的准确度（相对回收率为 95.7-98.3%）。大黄酸、大黄酚和灵芝的平均血浆蛋白结合率分别为96.7%、41.8%和46.7%。结果表明，所提出的程序不仅可以提取和浓缩大黄中的蒽醌，还可以确定其血浆蛋白结合率。4 ng/mL，富集因子在 54.2–100.7 范围内。线性为 0.5–500 ng/mL，r2 ≥ 0.9838。还获得了令人满意的准确度（相对回收率为 95.7-98.3%）。大黄酸、大黄酚和灵芝的平均血浆蛋白结合率分别为96.7%、41.8%和46.7%。结果表明，所提出的程序不仅可以提取和浓缩大黄中的蒽醌，还可以确定其血浆蛋白结合率。4 ng/mL，富集因子在 54.2–100.7 范围内。线性为 0.5–500 ng/mL，r2 ≥ 0.9838。还获得了令人满意的准确度（相对回收率为 95.7-98.3%）。大黄酸、大黄酚和灵芝的平均血浆蛋白结合率分别为96.7%、41.8%和46.7%。结果表明，所提出的程序不仅可以提取和浓缩大黄中的蒽醌，还可以确定其血浆蛋白结合率。