Chemical Works of Gedeon Richter Plc., API Research and Development - Spectroscopic Research, Budapest, Hungary.
J Pharm Biomed Anal. 2013 Oct;84:293-308. doi: 10.1016/j.jpba.2012.08.019. Epub 2012 Aug 28.
In the course of exploring the possibilities of developing a new, improved process at Gedeon Richter for the production of the "bisindole" alkaloids vinblastine (VLB) and vincristine (VCR), some novel VLB/VCR-related trace impurities were detected by analytical HPLC. Following isolation by preparative HPLC, a combination of 1D and 2D ultra high-field NMR and high-resolution (HR) (LC-)MS/MS studies allowed the structural identification and complete spectral characterization of several hitherto unpublished VLB/VCR-analogue impurities. Since the impurities could not be isolated in entirely pure forms and were available only in minute, mass-limited quantities, accessing the spectral information needed for their ab initio structure determination was met with various practical difficulties. Successful structure determination therefore relied heavily on the availability and use of detailed and definitive spectral data for both VLB and VCR. In particular, the utilization of detailed (1)H, (13)C, and (15)N NMR assignments as well as (1)H-(1)H, (1)H-(13)C and (1)H-(15)N spin-spin connectivities pertaining to different solvents for VLB/VCR base and sulphate salt was required. Although NMR studies on VLB base and other bisindoles were reported earlier in the literature, an NMR characterization of VLB and VCR under the above-mentioned circumstances and using ultra-high field instrumentation is either scarcely available or entirely lacking, therefore the necessary data had to be obtained in-house. Likewise, a modern tandem HR-ESI-MS/MS(n) fragmentation study of VLB and VCR has not been published yet. In the present paper we therefore give a thorough NMR and MS characterization of VLB and VCR specifically with a view to filling this void and to provide sufficiently extensive and solid reference data for the structural investigation of the aforementioned VLB/VCR impurities. Besides being scientifically relevant in its own right, the disclosed data should be useful for anyone interested in VLB/VCR-related molecules at a structural level.
在探索盖德恩·里希特（Gedeon Richter）生产“双吲哚”生物碱长春碱（VLB）和长春新碱（VCR）的新工艺的可能性的过程中，通过分析 HPLC 检测到了一些新型的 VLB/VCR 相关痕量杂质。通过制备 HPLC 分离后，结合一维和二维超高场 NMR 和高分辨率（HR）（LC-）MS/MS 研究，对几种以前未发表的 VLB/VCR 类似物杂质进行了结构鉴定和全谱特征描述。由于杂质不能以完全纯的形式分离，并且只能获得微量的质量限制的数量，因此获取用于从头确定其结构的光谱信息遇到了各种实际困难。因此，成功的结构确定在很大程度上依赖于 VLB 和 VCR 的详细和明确的光谱数据的可用性和使用。特别是，需要利用详细的（1）H、（13）C 和（15）N NMR 分配以及（1）H-（1）H、（1）H-（13）C 和（1）H-（15）N 自旋-自旋连接性，这些涉及 VLB/VCR 碱和硫酸盐盐的不同溶剂。尽管以前在文献中报道了 VLB 碱和其他双吲哚的 NMR 研究，但在上述情况下使用超高场仪器对 VLB 和 VCR 进行 NMR 表征是很少见的，或者完全没有，因此必须在内部获得必要的数据。同样，VLB 和 VCR 的现代串联 HR-ESI-MS/MS（n）碎裂研究尚未公布。在本文中，我们对 VLB 和 VCR 进行了全面的 NMR 和 MS 表征，特别是为了填补这一空白，并为上述 VLB/VCR 杂质的结构研究提供足够广泛和可靠的参考数据。除了本身具有科学意义外，所公开的数据对于任何对结构水平的 VLB/VCR 相关分子感兴趣的人都应该是有用的。
