A novel LC-MS method using collagen marker peptides for species identification of glue applicable to samples with multiple animal origins
A novel LC-MS method using collagen marker peptides for species identification of glue applicable to samples with multiple animal origins
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DOI：
10.1186/s40494-018-0209-y
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发表时间：
2018-07-13
期刊：
影响因子：
2.5
通讯作者：
Hattori, Shunji
中科院分区：
文献类型：
--
作者：
Kumazawa, Yuki;Taga, Yuki;Hattori, Shunji
来源于富含胶原蛋白的组织的动物胶被广泛用作艺术品中的粘合剂和粘合剂。确定胶水的动物来源不仅对了解艺术技术和历史背景很重要，而且对艺术品的修复和保护进行适当的处理也很重要。最近，质谱方法已被用于胶的种类鉴定。然而，现有的方法似乎不适用于含有多种动物来源的样品，并且由于公共数据库中胶原蛋白的序列信息不完整，靶物种的范围受到限制。在本研究中，我们建立了一种新的方法来区分胶水从八种动物适用于样品与多种动物来源。采用LC-MS多反应监测模式对胰蛋白酶消化后的胶样进行分析，并根据12种I型胶原源性标志肽的检测模式确定动物来源。使用数据库独立的方法，我们成功地确定了商业产品的动物来源，并发现一些胶水的申报物种是不正确的。此外，一些产品被确定含有两种不同的动物来源。使用标记肽估计了几种不纯胶中动物来源的相对丰度，这有助于推测检测到多个物种的原因。我们分析了一幅油画（Camille Pissarro的《收获》，1882年），并澄清了画布的底层和尺寸层中使用的胶水来自牛（类似于65%）和羊（类似于35%）。
Animal glues derived from collagen-rich tissue are widely used as adhesives and binders in artworks. Identification of the animal source of glue is important not only to understand the artistic technique and historical background, but also to conduct appropriate treatment for restoration and conservation of artworks. Recently, mass spectrometric approaches have been used for species identification of glue. However, it appears that the existing methods were not applied to samples containing multiple animal sources, and the range of target species has been limited because of incomplete sequence information of collagen in public databases. In the present study, we established a novel method for discrimination of glues from eight animals applicable to samples with multiple animal origins. Trypsin-digested glue samples were analyzed by LC-MS in multiple reaction monitoring mode, and the animal source was determined based on the detection patterns of 12 type I collagen-derived marker peptides. Using the database-independent method, we successfully identified the animal source of commercial products and found the declared species for some glues to be incorrect. In addition, some products were identified to contain two different animal origins. Relative abundance of the animal origins in several impure glues was estimated using the marker peptides, which helped to speculate the reason for the detection of multiple species. We analyzed a painting (The Harvest by Camille Pissarro, 1882) and clarified that glues used in the ground and size layers of the canvas were derived from cattle (similar to 65%) and sheep (similar to 35%).