Colombo Erika, Mauri Lucio, Marinozzi Maria, Rudd Timothy R, Yates Edwin A, Ballabio Davide, Guerrini Marco
Institute for Chemical and Biochemical Research G. Ronzoni, via G. Colombo 81, 20133 Milan, Italy.
National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire, UK; Department of Biochemistry and Systems Biology, ISMIB, University of Liverpool, Liverpool L69 7ZB United Kingdom.
J Pharm Biomed Anal. 2022 May 30;214:114724. doi: 10.1016/j.jpba.2022.114724. Epub 2022 Mar 12.
Heparin has been used successfully as a clinical antithrombotic for almost one century. Its isolation from animal sources (mostly porcine intestinal mucosa) involves multistep purification processes starting from the slaughterhouse (as mucosa) to the pharmaceutical plant (as the API). This complex supply chain increases the risk of contamination and adulteration, mainly with non-porcine ruminant material. The structural similarity of heparins from different origins, the natural variability of the heparin within samples from each source as well as the structural changes induced by manufacturing processes, require increasingly sophisticated methods capable of detecting low levels of contamination. The application of suitable multivariate classification approaches on API H NMRspectra serve as rapid and reliable tools for product authentication and the detection of contaminants. Soft Independent Modeling of Class Analogies (SIMCA), Discriminant Analysis (DA), Partial Least Square Discriminant Analysis (PLS-DA) and local classification methods (kNN, BNN and N3) were tested on about one hundred certified heparin samples produced by 14 different manufacturers revealing that Partial Least Squares Discriminant Analysis (PLS-DA) provided the best discrimination of contaminated batches, with a balanced accuracy of 97%.
肝素作为一种临床抗血栓药物已成功使用了近一个世纪。从动物来源（主要是猪肠黏膜）分离肝素涉及从屠宰场（作为黏膜）到制药厂（作为原料药）的多步纯化过程。这种复杂的供应链增加了污染和掺假的风险，主要是与非猪反刍动物材料有关。不同来源肝素的结构相似性、每个来源样品中肝素的自然变异性以及制造过程引起的结构变化，都需要越来越复杂的方法来检测低水平的污染。在原料药1H NMR光谱上应用合适的多变量分类方法可作为产品鉴定和污染物检测的快速可靠工具。对14家不同制造商生产的约100个经认证的肝素样品测试了类类比软独立建模（SIMCA）、判别分析（DA）、偏最小二乘判别分析（PLS-DA）和局部分类方法（kNN、BNN和N3），结果表明偏最小二乘判别分析（PLS-DA）对受污染批次的判别效果最佳，平衡准确率为97%。
