Abstract Lettuce (Lactuca sativa) and its bioactive constituent lactucaxanthin are gaining importance due to their antidiabetic property. Hence, aim of this study is to establish a simple purification technique for lactucaxanthin and its antioxidant property, cytotoxicity, and cellular uptake in 3T3-L1 cells (pre-adipocytes). Results revealed that lactucaxanthin is higher (mg/100 g dry weight) in green lettuce (3.05 ± 0.12) than Romania (0.42 ± 0.05) and Iceberg (0.36 ± 0.06), respectively. The purity of lactucaxanthin separated by TLC followed by preparatory HPLC is 99 ± 1% with λmax 439 nm. FTIR, LC-MS (M + H+‒H2O = 551) and NMR analysis revealed the characteristic chemical structure of lactucaxanthin. Cytotoxicity of lactucaxanthin in 3T3-L1 cells treated at 1, 3, 5, 10, 30, 50, 60, 100 μM for 24 hr revealed 86–89% cell viability up to 10 µM concentration. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS)-mediated radical scavenging (35.65 ± 0.85 µg/ml and 4.92 ± 0.81 µg/ml), total antioxidant 5 µg (equivalent to 23.71 ± 1.88 ascorbic acid) activity and reductive capability 20 µg (26.14 µg quercetin equivalent) of lactucaxanthin proved its antioxidant potential. 3T3-L1 cellular uptake of lactucaxanthin was found time dependent with absorption maxima of 46.35 ng after 24 hr treatment (10 µM). The present results help in effective positioning of lactucaxanthin for food and pharma application to fight against diabetes and dyslipidemia. Graphical Abstract

中文翻译：

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从生菜（Lactuca sativa）中纯化并通过 LC-MS 和 NMR 表征的乳黄质的细胞毒性和 3T3-L1 细胞摄取

摘要 莴苣 (Lactuca sativa) 及其生物活性成分乳黄素因其抗糖尿病特性而越来越重要。因此，本研究的目的是建立一种简单的乳黄素纯化技术及其在 3T3-L1 细胞（前脂肪细胞）中的抗氧化特性、细胞毒性和细胞摄取。结果显示，绿色生菜 (3.05 ± 0.12) 中的乳黄质素 (mg/100 g 干重) 分别高于罗马尼亚 (0.42 ± 0.05) 和冰山 (0.36 ± 0.06)。TLC 和制备型 HPLC 分离的乳黄素纯度为 99 ± 1%，λmax 439 nm。FTIR、LC-MS (M + H+-H2O = 551) 和 NMR 分析揭示了乳黄素的特征化学结构。在 1, 3, 5, 10, 30, 50, 60, 3T3-L1 细胞中乳黄素的细胞毒性，100 μM 24 小时显示高达 10 μM 浓度的 86–89% 细胞活力。2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS)-介导的自由基清除 (35.65 ± 0.85 µg/ml 和 4.92 ± 0.81 µg/ ml)，总抗氧化剂 5 µg（相当于 23.71 ± 1.88 抗坏血酸）活性和还原能力 20 µg（26.14 µg 槲皮素当量）的乳黄黄素证明了其抗氧化潜力。发现乳黄黄素的 3T3-L1 细胞摄取时间依赖于吸收最大值为处理 24 小时后为 46.35 ng (10 µM)。目前的结果有助于有效定位乳黄质在食品和药物中的应用，以对抗糖尿病和血脂异常。图形摘要 发现 3T3-L1 细胞对乳黄素的吸收具有时间依赖性，在 24 小时处理 (10 µM) 后吸收最大值为 46.35 ng。目前的结果有助于有效定位乳黄质在食品和药物中的应用，以对抗糖尿病和血脂异常。图形概要 发现 3T3-L1 细胞对乳黄素的吸收具有时间依赖性，在 24 小时处理 (10 µM) 后吸收最大值为 46.35 ng。目前的结果有助于有效定位乳黄质在食品和药物中的应用，以对抗糖尿病和血脂异常。图形概要