This study focuses on the extraction of bioactive compounds from date seeds using five polyol-based deep eutectic solvents (P-DESs) in combination with hybrid green extraction techniques, specifically microwave-assisted extraction (MAE), homogenization-assisted extraction (HAE), and ultrasound-assisted extraction (UAE). The optimization of these extraction techniques was achieved using P-DESs showing the highest efficiency for extracting date seed bioactive compounds using response surface methodology (RSM) and central composite design (CCD) approach. The optimized conditions from three green techniques were further applied in the form of hybrid green extraction techniques, involving six binary and three ternary methods, to assess the percentage increase in the extraction efficiency of date seed bioactive polyphenolics. Among the five P-DESs tested, choline chloride: ethylene glycol (ChCl:Eg) exhibited the highest extraction efficiency for recovering date seed phenolic compounds. Using ChCl:Eg as the P-DES, the highest extraction efficacy was found with MAE, followed by > HAE and > UAE. In addition, all hybrid extraction techniques showed higher extraction efficiencies than the single extraction methods. Notably, the binary hybrid techniques combining UAE and MAE (UMAE), HAE and MAE (HMAE) resulted in significantly higher recovery of bioactive compounds, with 52 % and 49 % increases in total phenolic content, respectively, compared to single extraction techniques. The lowest MIC and MBC of P-DES (ChCl:Eg) and date seed P-DES based extract recorded against all the tested bacterial strains was 40 % and 20 % respectively. Furthermore, the date seed extract from MAE was used to extend the shelf life of Oreochromis niloticus stored at 4 °C for 10 days. The results indicated that the date seed polyphenolic extract effectively inhibited microbial growth in Oreochromis niloticus during refrigerated storage, with the total bacterial count (TBC) of all the treated samples within the recommended acceptability limit of < 6 log CFU/g compared to the untreated samples, which showed a total bacterial count (TBC) > 6 log CFU/g. This study demonstrated that sequential hybrid techniques enhance and intensify the recovery of bioactive compounds more effectively than any single green technique.

中文翻译：

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序列绿色杂交技术（超声、微波和高剪切均质）提取枣籽生物活性化合物及其作为添加剂延长尼罗河鱼保质期的比较研究


本研究的重点是使用五种基于多元醇的共熔溶剂 （P-DES） 结合混合绿色提取技术，特别是微波辅助提取 （MAE）、均质辅助提取 （HAE） 和超声辅助提取 （UAE），从枣籽中提取生物活性化合物。这些提取技术的优化是使用 P-DES 实现的，显示出使用响应面法 （RSM） 和中心复合设计 （CCD） 方法提取枣籽生物活性化合物的效率最高。将 3 种绿色技术的优化条件进一步应用于杂交绿色提取技术，包括 6 种二元和 3 种三元方法，以评估枣籽生物活性多酚提取效率的百分比增加。在测试的 5 种 P-DES 中，氯化胆碱：乙二醇 （ChCl：Eg） 对回收枣籽酚类化合物的提取效率最高。使用 ChCl：Eg 作为 P-DES，MAE 的提取效率最高，其次是 > HAE 和 > UAE。此外，所有杂交提取技术都显示出比单一提取方法更高的提取效率。值得注意的是，与单一提取技术相比，结合 UAE 和 MAE （UMAE）、HAE 和 MAE （HMAE） 的二元混合技术显著提高了生物活性化合物的回收率，总酚含量分别增加了 52% 和 49%。在所有测试的细菌菌株中记录的 P-DES （ChCl：Eg） 和基于枣种子 P-DES 的提取物的最低 MIC 和 MBC 分别为 40% 和 20%。此外，来自 MAE 的枣籽提取物用于延长 Oreochromis niloticus 在 4 °C 下储存 10 天的保质期。 结果表明，枣籽多酚提取物在冷藏过程中有效抑制了尼罗河毒株的微生物生长，所有处理样品的总细菌计数 （TBC） 均在推荐的可接受限度 < 6 log CFU/g 内，而未处理样品则显示总细菌计数 （TBC） > 6 log CFU/g。这项研究表明，顺序混合技术比任何单一的绿色技术都更有效地增强和加强生物活性化合物的回收。