Gotti R, Fiori J, Bartolini M, Cavrini V
Dipartimento di Scienze Farmaceutiche, Alma Mater Studiorum Università di Bologna, Via Belmeloro 6, 40126 Bologna, Italy.
J Pharm Biomed Anal. 2006 Sep 11;42(1):17-24. doi: 10.1016/j.jpba.2006.01.003. Epub 2006 Feb 7.
Amaryllidaceae are known as ornamental plants, furthermore some species of this family contain galanthamine, an acetylcholinesterase inhibitor approved for the treatment of Alzheimer's disease, and other alkaloids with interesting pharmacological activity. In the present work, the quali- and quantitative analysis of Amaryllidaceae-type alkaloids in the bulbs of Narcissus species is presented using different analytical approaches. Extracts of Narcissus pseudonarcissus cv. Carlton and Narcissus jonquilla Quail, were first examined by GC-MS using a Rtx-5 MS (programmed temperature) and the major alkaloids were identified. Together with galanthamine, high contents of haemanthamine, were found. Galanthamine was reliably quantified by GC-MS, whereas haemanthamine partly decomposed under the GC conditions, thus alternative analytical methods were investigated. Firstly, reversed-phase HPLC-ESI-MS was applied to identify and isolate at semipreparative levels haemanthamine. The compound was fully characterized by MS/MS and (1)H NMR and then used as a reference substance. The quantitation of both galanthamine and haemanthamine was then accomplished by capillary electrophoresis with spectrophotometric detection. A non-aqueous (NACE) approach was selected in order to use a running buffer fully compatible with samples in organic solvent. In particular, a mixture methanol-acetonitrile (75:25, v/v) containing ammonium acetate (90 mM) was used as a background electrolyte. The same analytical sample was subjected to GC-MS and NACE analysis; the different selectivity displayed by these techniques allowed different separation profiles that can be useful in phytochemical characterization of the extracts. The GC-MS and NACE methods were validated and applied to the quantitation of galanthamine (GC-MS and NACE) and haemanthamine (NACE) in bulbs of N. jonquilla.
石蒜科植物以观赏植物而闻名，此外，该科的一些物种含有加兰他敏，一种被批准用于治疗阿尔茨海默病的乙酰胆碱酯酶抑制剂，以及其他具有有趣药理活性的生物碱。在本研究中，使用不同的分析方法对水仙属植物鳞茎中的石蒜科型生物碱进行了定性和定量分析。首先，采用GC-MS（程序升温），使用Rtx-5 MS对水仙‘卡尔顿’和水仙‘鹌鹑’的提取物进行检测，并鉴定出主要生物碱。除加兰他敏外，还发现了高含量的海芒果碱。加兰他敏可通过GC-MS可靠地定量，而海芒果碱在GC条件下部分分解，因此研究了替代分析方法。首先，应用反相HPLC-ESI-MS在半制备水平上鉴定和分离海芒果碱。该化合物通过MS/MS和¹H NMR进行了全面表征，然后用作参考物质。然后通过毛细管电泳-分光光度检测法对加兰他敏和海芒果碱进行定量。选择了非水（NACE）方法，以便使用与有机溶剂中的样品完全兼容的运行缓冲液。具体而言，使用含有醋酸铵（90 mM）的甲醇-乙腈混合物（75:25，v/v）作为背景电解质。对同一分析样品进行了GC-MS和NACE分析；这些技术显示出的不同选择性允许得到不同的分离图谱，这在提取物的植物化学表征中可能是有用的。GC-MS和NACE方法经过验证，并应用于水仙鳞茎中加兰他敏（GC-MS和NACE）和海芒果碱（NACE）的定量分析。
