Tessema Efrem N, Gebre-Mariam Tsige, Schmelzer Christian E H, Neubert Reinhard H H
Department of Pharmaceutical Technology and Biopharmaceutics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Str. 4, 06120 Halle Saale, Germany.
Department of Pharmaceutics and Social Pharmacy, School of Pharmacy, College of Health Sciences, Addis Ababa University, P.O. Box 1176, Addis Ababa, Ethiopia.
J Pharm Biomed Anal. 2017 Jul 15;141:241-249. doi: 10.1016/j.jpba.2017.04.036. Epub 2017 Apr 24.
Chronic skin conditions such as atopic dermatitis, psoriasis and aged skin are characterized by defective skin barrier and dryness which are associated with reduced levels of skin ceramides (CERs). The beneficial effects of plant-derived CERs for skin hydration and skin barrier recovery have been shown in several studies. Although plenty of glucosylceramide (GlcCER)-based dietary supplements meant for skin barrier improvement have been marketed, there are limited commercial sources of plant GlcCERs. In an attempt to explore alternative GlcCER sources, a reversed phase LC-MS/MS method with atmospheric pressure chemical ionization (APCI) interface was developed for separation and structural identification of GlcCERs isolated from three plants. The GlcCERs were extracted from the seeds of grass pea (Lathyrus sativus L.), Ethiopian mustard (Brassica carinata) and haricot bean (Phaseolus vulgaris) and purified by column chromatography and preparative LC-MS. The individual GlcCER species were further separated and qualitatively analyzed by LC/APCI-MS/MS. The amount of GlcCERs in each plant was quantified by HPTLC. All GlcCER species detected in the three plants consisted of C18 di/trihydroxy sphingoid bases amide linked with hydroxy fatty acids (C14-C24). The trihydroxy SBs were acylated with very long chain FAs (C22-C24). The major GlcCERs derived from grass pea, Ethiopian mustard and haricot bean are composed of sphingenine (d18:1) linked to hydroxypalmitic acid (h16:0), 4-hydroxy-8-sphingenine (t18:1) coupled with hydroxynervonic acid (h24:1) and sphingadienine (d18:2) joined with h16:0, respectively. The GlcCERs contents in haricot bean (161.2mg/kg) and grass pea (130.0mg/kg) were found to be higher compared to Ethiopian mustard (71.8mg/kg). This qualitative and quantitative information suggests that the two plants of the Fabaceae family (haricot bean and grass pea) are potential alternative sources of GlcCERs for their use in products meant for the recovery of skin barrier function. The LC/APCI-MS/MS method described here has proven to be reliable for the screening of other potential plants containing GlcCERs.
慢性皮肤疾病，如特应性皮炎、牛皮癣和老年皮肤，其特征是皮肤屏障功能缺陷和干燥，这与皮肤神经酰胺（CERs）水平降低有关。多项研究表明，植物来源的神经酰胺对皮肤保湿和皮肤屏障修复具有有益作用。尽管市面上已有大量旨在改善皮肤屏障的基于葡萄糖神经酰胺（GlcCER）的膳食补充剂，但植物GlcCER的商业来源有限。为了探索替代的GlcCER来源，开发了一种带有大气压化学电离（APCI）接口的反相LC-MS/MS方法，用于分离和结构鉴定从三种植物中分离出的GlcCER。从草豌豆（Lathyrus sativus L.）、埃塞俄比亚芥菜（Brassica carinata）和菜豆（Phaseolus vulgaris）的种子中提取GlcCER，并通过柱色谱和制备型LC-MS进行纯化。通过LC/APCI-MS/MS进一步分离并定性分析各个GlcCER种类。通过HPTLC对每种植物中GlcCER的含量进行定量。在这三种植物中检测到的所有GlcCER种类均由与羟基脂肪酸（C14-C24）酰胺连接的C18二/三羟基鞘氨醇碱组成。三羟基鞘氨醇碱被极长链脂肪酸（C22-C24）酰化。源自草豌豆、埃塞俄比亚芥菜和菜豆的主要GlcCER分别由与羟基棕榈酸（h16:0）连接的鞘氨醇（d18:1）、与羟基神经酸（h24:1）偶联的4-羟基-8-鞘氨醇（t18:1）和与h16:0连接的鞘氨二烯（d18:2）组成。发现菜豆（161.2mg/kg）和草豌豆（130.0mg/kg）中的GlcCER含量高于埃塞俄比亚芥菜（71.8mg/kg）。这种定性和定量信息表明，豆科的两种植物（菜豆和草豌豆）是GlcCER的潜在替代来源，可用于旨在恢复皮肤屏障功能的产品中。此处描述的LC/APCI-MS/MS方法已被证明对于筛选其他含有GlcCER的潜在植物是可靠的。
