Department of Organic Chemistry, Faculty of Chemistry and Technology, University of Split, Ruđera Boškovića 35, 21000 Split, Croatia.
School of Medicine, University of Split, Šoltanska 2, 21000 Split, Croatia.
Biomolecules. 2020 Feb 1;10(2):215. doi: 10.3390/biom10020215.
Glucosinolates (GSLs) from L. seeds were analyzed qualitatively and quantitatively by their desulfo counterparts using UHPLC-DAD-MS/MS technique and by their volatile breakdown products, isothiocyanates (ITCs), using GC-MS technique. GSL breakdown products were obtained by conventional techniques (hydrodistillation in a Clevenger type apparatus (HD), CHCl extraction after myrosinase hydrolysis (EXT) for 24 h) as well as by modern techniques, microwave-assisted distillation (MAD) and microwave hydrodiffusion and gravity (MHG). Seven GSLs were identified as follows: isopropyl GSL (), -butyl GSL (), 5-(methylsulfinyl)pentyl GSL (), 6-(methylsulfinyl)hexyl GSL (), 5-(methylsulfanyl)pentyl GSL (), 6-(methylsulfanyl)hexyl GSL (), and benzyl GSL (). Additionally, pent-4-enyl- and hex-5-enyl ITCs were detected in the volatile extracts. However, their corresponding GSLs were not detected using UHPLC-DAD-MS/MS. Thus, they are suggested to be formed during GC-MS analysis via thermolysis of 5-(methylsulfinyl)pentyl- and 6-(methylsulfinyl)hexyl ITCs, respectively. Volatile isolates were tested for their cytotoxic activity using MTT assay. EXT and MHG showed the best cytotoxic activity against human lung cancer cell line A549 during an incubation time of 72 h (IC 18.8, and 33.5 μg/mL, respectively), and against breast cancer cell line MDA-MB-231 after 48 h (IC 6.0 and 11.8 μg/mL, respectively). These activities can be attributed to the ITCs originating from and .
利用超高效液相色谱-二极管阵列-串联质谱联用（UHPLC-DAD-MS/MS）技术和气相色谱-质谱联用（GC-MS）技术，通过其脱硫对应物和挥发性分解产物——异硫氰酸酯（ITCs）对 L.种子中的硫代葡萄糖苷（GSLs）进行定性和定量分析。通过传统技术（在 Clevenger 型装置中进行水蒸馏（HD），在经过辣根过氧化物酶水解（EXT）24 小时后用 CHCl3 萃取（EXT））以及现代技术（微波辅助蒸馏（MAD）和微波水扩散与重力（MHG））获得 GSL 分解产物。鉴定出 7 种 GSLs，分别为：异丙基 GSL（）、-丁基 GSL（）、5-（甲硫基）戊基 GSL（）、6-（甲硫基）己基 GSL（）、5-（甲基亚磺酰基）戊基 GSL（）、6-（甲基亚磺酰基）己基 GSL（）和苄基 GSL（）。此外，在挥发性提取物中还检测到戊-4-烯基和己-5-烯基 ITCs。然而，使用 UHPLC-DAD-MS/MS 并未检测到相应的 GSLs。因此，推测它们是通过 5-（甲硫基）戊基-和 6-（甲硫基）己基 ITCs 的热解分别在 GC-MS 分析中形成的。通过 MTT 测定法对挥发性分离物的细胞毒性活性进行测试。在 72 小时孵育时间内，EXT 和 MHG 对人肺癌细胞系 A549 显示出最佳的细胞毒性活性（IC 50 分别为 18.8 和 33.5 μg/mL），对乳腺癌细胞系 MDA-MB-231 在 48 小时孵育时间内显示出最佳的细胞毒性活性（IC 50 分别为 6.0 和 11.8 μg/mL）。这些活性可以归因于源自 和 的 ITCs。
