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采用超声辅助从种子中提取富含甾醇和四环三萜类化合物的脂质,并运用气相色谱/质谱联用技术进行脂质组学分析。

Ultrasound-assisted lipid extractions, enriched with sterols and tetranortriterpenoids, from seeds and the application of lipidomics using GC/MS.

作者信息

Silva Dos Reis André, Santos Alberdan Silva, Francisco de Carvalho Gonçalves José

机构信息

Laboratory of Systematic Investigation in Biotechnology and Molecular Biodiversity, Federal University of Pará (UFPA) Belém PA 66075-110 Brazil

Laboratory of Plant Physiology and Biochemistry, National Institute of Amazonian Research (MCTI-INPA) Manaus AM 69060-020 Brazil.

出版信息

RSC Adv. 2021 Oct 8;11(52):33160-33168. doi: 10.1039/d1ra04776k. eCollection 2021 Oct 4.

Abstract

This study describes the optimized stages of lipid extraction assisted by ultrasound to increase the concentrations of limonoids and steroids from andiroba seeds, identified as Aublet, and the lipidome analyzed by TLC and GC/MS. After boiling and peeling, crushed seeds were subjected to extractions with -hexane (G1), acetone (G2), and methanol (G3) in an ultrasonic bath for 30 minutes at 50 °C. These extracts were analyzed by analytical TLC; aliquots of each extract and a marketable oil, used as a reference, were transesterified followed by silylation with BSTFA + 1% TMCS, and analyzed by GC/MS. The reference oil and the biomass of the seeds did not present significant differences in the profiles of free fatty acids (FFA). However, differences were observed in the profiles of tetranortriterpenoids (TnTT) (limonoids) in the seeds. Afterward, the analysis of the extracts G1, G2, and G3 detected free steroids: campesterol, stigmasterol, and β-sitosterol; and limonoids: 7-deacetyl-7-oxogedunin, 6α-acetoxy-gedunin, deacetylgedunin, and epoxyacetylgedunin. The use of lipidomic techniques associated with ultrasound-assisted extraction was applied for the first time to enhance the triterpenoids and steroids, mainly in G3, describing a faster and more economical process, and allowing a one-step lipidome analysis of the andiroba seeds.

摘要

本研究描述了超声辅助脂质提取的优化阶段,以提高从被鉴定为奥布莱特的印加果种子中柠檬苦素和甾体的浓度,并通过薄层色谱(TLC)和气相色谱/质谱联用(GC/MS)分析脂质组。经过煮沸和去皮后,将碾碎的种子在50℃的超声浴中用正己烷(G1)、丙酮(G2)和甲醇(G3)萃取30分钟。这些提取物通过分析型TLC进行分析;将每种提取物的等分试样和一种市售油作为参比进行酯交换,然后用N,O-双(三甲基硅基)三氟乙酰胺(BSTFA)+1%三甲基氯硅烷(TMCS)进行硅烷化,并通过GC/MS进行分析。参比油和种子生物质在游离脂肪酸(FFA)谱方面没有显著差异。然而,在种子的四降三萜类化合物(TnTT)(柠檬苦素)谱方面观察到了差异。随后,对提取物G1、G2和G3的分析检测到了游离甾体:菜油甾醇、豆甾醇和β-谷甾醇;以及柠檬苦素:7-脱乙酰基-7-氧代格杜宁、6α-乙酰氧基格杜宁、脱乙酰格杜宁和环氧乙酰格杜宁。脂质组学技术与超声辅助提取相结合的方法首次被用于提高三萜类化合物和甾体的含量,主要是在G3中,描述了一种更快、更经济的方法,并允许对印加果种子进行一步脂质组分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/77f7/9042240/3390ca7e9c49/d1ra04776k-f1.jpg

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