用超声波辅助从 Tubocapsicum anomalum 中萃取山奈酚内酯:工艺优化、分离和鉴定以及抗增殖活性。

IF 8.7 1区 化学 Q1 ACOUSTICS Ultrasonics Sonochemistry Pub Date : 2024-09-02 DOI:10.1016/j.ultsonch.2024.107052
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The optimal extraction conditions were determined as follows: the liquid–solid ratio was 20:1, the extraction solvent was 70 % ethanol, the ultrasonic power was 250 W, the ultrasonic time was 40 min, and the ultrasonic temperature was 50 °C. Under these conditions, the average yields of tubocapsenolide A (Te-A) and tubocapsanolide A (Ta-A) can reach 2.87 ± 0.12 mg/g and 1.18 ± 0.05 mg/g, respectively. We further compared extraction rates of two withanolides from different parts of <em>T. anomalum</em> using ultrasonic and traditional extraction methods. Ultrasonic extraction significantly increased rates, with the highest yields from leaves, followed by stems and fruits. The results show that ultrasonic optimization can improve extraction rate, reduce time, lower costs, enhance quality, and increase yield. Therefore, the optimized ultrasonic-assisted extraction process was adopted to extract the aerial parts of <em>T. anomalum</em> and separate the components. 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引用次数: 0

摘要

薯蓣皂苷作为一种富含抗肿瘤薯蓣皂苷的中药植物,需要高效的提取方法。本文首先建立了高效液相色谱法检测岩白菜素内酯的方法,并采用甲醇-水溶剂系统进行梯度洗脱。研究发现,反式蘑菇叶片中的山奈酚苷含量最高,其次是茎和果实,根中几乎没有含量。在实际采摘过程中,采集到的叶片数量相对较少,而茎的数量最多。因此,采用方框-贝肯响应面法对超声波辅助提取异株鳖甲茎中的香叶醇苷的过程进行了优化。最佳提取条件确定如下:液固比为 20:1,提取溶剂为 70%乙醇,超声功率为 250 W,超声时间为 40 min,超声温度为 50 °C。在此条件下,管花内酯 A(Te-A)和管花内酯 A(Ta-A)的平均产率分别为 2.87 ± 0.12 mg/g 和 1.18 ± 0.05 mg/g。我们进一步比较了用超声波提取法和传统提取法从 T. anomalum 不同部位提取两种岩白菜素内酯的比率。超声波萃取法明显提高了萃取率,其中叶的萃取率最高,其次是茎和果实。结果表明,优化超声波提取法可以提高提取率、缩短时间、降低成本、提高质量和增加产量。因此,采用优化后的超声波辅助提取工艺来提取蚂蚁草的气生部分并分离其成分。经过优化后,提取物经过多次色谱分离,除了 15 种已知化合物(11-25)外,还分离出 8 种以前未曾描述过的岩白菜素内酯(1-8)和 2 种人工岩白菜素内酯(9-10)。通过大量光谱数据分析,确定了这些化合物的结构。评估了这些化合物对多种癌细胞系的抗增殖作用,包括人肝癌细胞(HepG2、Hep3B 和 MHCC97-H)、人肺癌细胞(A549)、人纤维肉瘤癌细胞(HT1080)、人慢性粒细胞白血病细胞(K562)和人乳腺癌细胞(MDA-MB-231 和 MCF7)。化合物 1-3、5、7、11、13、15-16 和 22 显示出显著的活性,IC50 值为 5.14-19.87 μM。上述结果表明,利用超声波辅助萃取技术可以更有效地从蚂蚁草中获得新的岩白菜素内酯,从而提高蚂蚁草资源的利用率。
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Ultrasound-assisted extraction of withanolides from Tubocapsicum anomalum: Process optimization, isolation and identification, and antiproliferative activity

Tubocapsicum anomalum, a Chinese medicinal plant rich in anti-tumor withanolides, requires efficient extraction methods. In this paper, an HPLC method was first established for the detection of withanolides, and gradient elution was carried out using a methanol–water solvent system. It was found that the content of withanolides was the highest in the leaves of T. anomalum, followed by the stems and fruits, and almost none in the roots. During the actual picking process, the quantity of leaves collected was relatively small, while the number of stems was the highest. Therefore, the Box-Behnken response surface method was used to optimize the ultrasonic-assisted extraction process of withanolides from the stems of T. anomalum. The optimal extraction conditions were determined as follows: the liquid–solid ratio was 20:1, the extraction solvent was 70 % ethanol, the ultrasonic power was 250 W, the ultrasonic time was 40 min, and the ultrasonic temperature was 50 °C. Under these conditions, the average yields of tubocapsenolide A (Te-A) and tubocapsanolide A (Ta-A) can reach 2.87 ± 0.12 mg/g and 1.18 ± 0.05 mg/g, respectively. We further compared extraction rates of two withanolides from different parts of T. anomalum using ultrasonic and traditional extraction methods. Ultrasonic extraction significantly increased rates, with the highest yields from leaves, followed by stems and fruits. The results show that ultrasonic optimization can improve extraction rate, reduce time, lower costs, enhance quality, and increase yield. Therefore, the optimized ultrasonic-assisted extraction process was adopted to extract the aerial parts of T. anomalum and separate the components. After optimization, the extract underwent several chromatographic separations to isolate eight previously undescribed withanolides (18) and two artificial withanolides (910), in addition to fifteen known compounds (1125). Their structures were established through extensive spectroscopic data analysis. The compounds were evaluated for their antiproliferative effects against multiple cancer cell lines, including human hepatocellular carcinoma cells (HepG2, Hep3B, and MHCC97-H), human lung cancer cells (A549), human fibro-sarcoma cancer cells (HT1080), human chronic myeloid leukemia cells (K562), and human breast cancer cells (MDA-MB-231 and MCF7). Compounds 13, 5, 7, 11, 13, 1516, and 22 displayed significant activity with IC50 values of 5.14–19.87 μM. The above results indicate that ultrasonic-assisted extraction technology can be used to obtain new withanolides more efficiently from T. anomalum, thereby enhancing the utilization rate of T. anomalum resources.

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来源期刊
Ultrasonics Sonochemistry
Ultrasonics Sonochemistry 化学-化学综合
CiteScore
15.80
自引率
11.90%
发文量
361
审稿时长
59 days
期刊介绍: Ultrasonics Sonochemistry stands as a premier international journal dedicated to the publication of high-quality research articles primarily focusing on chemical reactions and reactors induced by ultrasonic waves, known as sonochemistry. Beyond chemical reactions, the journal also welcomes contributions related to cavitation-induced events and processing, including sonoluminescence, and the transformation of materials on chemical, physical, and biological levels. Since its inception in 1994, Ultrasonics Sonochemistry has consistently maintained a top ranking in the "Acoustics" category, reflecting its esteemed reputation in the field. The journal publishes exceptional papers covering various areas of ultrasonics and sonochemistry. Its contributions are highly regarded by both academia and industry stakeholders, demonstrating its relevance and impact in advancing research and innovation.
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