植物中1⁃脱氧野尻霉素特异性存在的考证

Research on the Specific Existence of 1⁃Deoxynojirimycin in Plants

1⁃脱氧野尻霉素(DNJ)是桑树中最具有特征性、标志性的生物碱,具有调节糖脂代谢、抗菌、抗病毒等药理活性,是一种极具开发价值的天然活性产物。目前DNJ检测方法主要为柱前衍生高效液相色谱⁃荧光检测法,该方法能够检测到所有与荧光剂结合生成的络合物,但对被检测的物质无法进行定性,往往造成假阳性。通过对桑科植物和已报道含有DNJ的非桑科植物进行检测分析,利用柱前衍生高效液相色谱⁃荧光检测法与高效液相色谱⁃质谱(HPLC⁃MS)检测法相结合,挖掘并考证含有DNJ的植物种类,为DNJ的开发利用提供更广泛的原料素材。检测发现,除了桑树,其他常见桑科植物菠萝蜜、全叶无花果、小叶榕、裂叶无花果、构树、大叶榕、橡皮树、柳叶榕中均不含有DNJ;非桑科植物也只有风信子中含有DNJ;常规桑品种强桑1号的桑叶DNJ含量是风信子叶片DNJ含量的212倍。结果表明,对于植物中DNJ的检测,应先通过HPLC⁃MS进行定性研究,然后再通过柱前衍生高效液相色谱⁃荧光检测法进行定量检测,以避免出现假阳性。

1⁃Deoxynojirimycin(DNJ)is the most characteristic and iconic alkaloid in mulberry.It has pharmacological activities such as regulating sugar and lipid metabolism,antibacterial and antiviral activities.It is a natural active product with great development value.At present,the main method for DNJ detection is pre⁃column derivatization high perform⁃ance liquid chromatography⁃fluorescence detection.This method can detect all complexes formed by binding with fluo⁃rescent agents,but it cannot qualitatively identify the substances being detected,which often results in false positives.By detecting and analyzing Moraceae plants and non⁃Moraceae plants that reported to contain DNJ,and combining pre⁃column derivatization high⁃performance liquid chromato⁃graphy⁃fluorescence detection with high⁃performance liquid chromatography⁃mass spectrometry(HPLC⁃MS)detection,we explored and verified the plant species containing DNJ,providing a wider range of raw materials for the development and utilization of DNJ.The test found that,except for mulberry tree(Morus L.),other common Moraceae plants such as Artocarpus heterophyllus Lam.,Ficus carica Linn.,Ficus concinna(Miq.)Miq.,Ficus cari⁃ca Linn.,Broussonetia papyrifera(L.)L′Hér.ex Vent.,Fi⁃cus altissima Blume,Ficus elastica Roxb.ex Hornem.,and Ficus celebensis Corner do not contain DNJ.Among non⁃Moraceae plants,only Hyacinthus orientalis L.Sp.Pl.contains DNJ.The DNJ content in the mulberry leaf of the conven⁃tional mulberry variety Qiangsang No.1 was 212 times that of Hyacinthus orientalis.The results showed that for the detec⁃tion of DNJ in plants,qualitative studies should be conducted by HPLC⁃MS first,followed by quantitative detection by pre⁃column derivatization HPLC⁃fluorescence detection to avoid false positives.