Spectral Service AG, Emil-Hoffmann-Straße 33, 50996 Cologne, Germany; Institute of Chemistry, Saratov State University,Astrakhanskaya Street 83, 410012 Saratov, Russia; Institute of Chemistry, Saint Petersburg State University, 13 B Universitetskaya Emb., St. Petersburg 199034, Russia.
Hemostasis and Thrombosis, Department of Pathology, Loyola University Medical Center, Maywood, IL, United States.
J Pharm Biomed Anal. 2018 May 10;153:168-174. doi: 10.1016/j.jpba.2018.02.041.
Nuclear magnetic resonance (NMR) spectroscopy is regarded as one of the most powerful and versatile analytical approaches to assure the quality of heparin preparations. In particular, it was recently demonstrated that by using H NMR coupled with chemometrics heparin and low molecular weight heparin (LMWH) samples derived from three major animal species (porcine, ovine and bovine) can be differentiated [Y.B. Monakhova et al. J. Pharm. Anal. 149 (2018) 114-119]. In this study, significant improvement of existing chemometric models was achieved by switching to 2D NMR experiments (heteronuclear multiple-quantum correlation (HMQC) and diffusion-ordered spectroscopy (DOSY)). Two representative data sets (sixty-nine heparin and twenty-two LMWH) belonged to different batches and distributed by different commercial companies were investigated. A trend for animal species differentiation was observed in the principal component analysis (PCA) score plot built based on the DOSY data. A superior model was constructed using HMQC experiments, where individual heparin (LMWH) clusters as well as their blends were clearly differentiated. The predictive power of different classification methods as well as unsupervised techniques (independent components analysis, ICA) clearly proved applicability of the model for routine heparin and LMWH analysis. The switch from 1D to 2D NMR techniques provides a wealth of additional information, which is beneficial for multivariate modeling of NMR spectroscopic data for heparin preparations.
核磁共振（NMR）光谱被认为是确保肝素制剂质量的最强大和最通用的分析方法之一。特别是，最近有人证明，通过使用 H NMR 结合化学计量学，可以区分来自三种主要动物物种（猪、羊和牛）的肝素和低分子量肝素（LMWH）样品[Y.B. Monakhova 等人。J. Pharm. Anal. 149（2018）114-119]。在这项研究中，通过切换到 2D NMR 实验（异核多量子相关（HMQC）和扩散有序光谱（DOSY）），对现有的化学计量学模型进行了重大改进。研究了两个具有代表性的数据集（69 个肝素和 22 个 LMWH），它们属于不同批次，并由不同的商业公司分发。基于 DOSY 数据构建的主成分分析（PCA）得分图中观察到动物物种分化的趋势。使用 HMQC 实验构建了一个优越的模型，其中单个肝素（LMWH）簇及其混合物得到了明显的区分。不同分类方法和无监督技术（独立成分分析，ICA）的预测能力清楚地证明了该模型适用于常规肝素和 LMWH 分析。从 1D 到 2D NMR 技术的转变提供了丰富的附加信息，这有利于肝素制剂 NMR 光谱数据的多元建模。
