组织工程医疗器械 利用液相色谱 串联质谱法定量牛I型胶原标志肽

[中文适用范围]： 本文件提供了使用液相色谱-串联质谱法（LC-MS/MS）对从牛组织中提取纯化的I型胶原标记肽进行定量的方法。本文件描述的方法旨在用于纯化牛I型胶原的标记肽检测，该胶原将用于构建组织工程医疗产品或基于胶原的其他生物材料，以进行产品质量控制。该方法也可用于对与其他动物来源和/或其他类型胶原混合的样品中的牛特异性及/或I型特异性胶原进行定性分析和定量检测。本文件不排除其他可能用于定量I型胶原的方法，例如可评估胶原总量的羟脯氨酸定量法。 注1：已知胶原至少有28种类型，且每种性质不同。本文件侧重于纯化牛I型胶原标记肽的定量。从皮肤、肌腱、骨骼等中分离的I型胶原可能含有其他类型胶原，例如III型和V型。I型胶原可来源于牛、猪等。对于其他类型胶原或来源于其他动物物种的I型胶原的定量，可使用本文件作为模板，但需要设计胶原类型特异性及/或动物物种特异性特征肽用于LC-MS/MS方法，并优化测定条件。 注2：对于结合了其他材料的支架的胶原标记肽定量，或组织ECM材料中包含的I型胶原或基于I型胶原的再生组织的定量，可参考本文件，但首先需要使用合理且经验证的方法（9,10）分离和/或纯化I型胶原，然后参考本文件提供的方法进行定量。 [外文原描述]： This document provides a method for quantification of marker peptide of type I collagen which was purified products extracted from bovine tissues with liquid chromatography - tandem mass spectrometry (LC-MS/MS). The method described in this document is intended to be used for marker peptide detection of purified bovine type I collagen which will be used for constructing tissue-engineered medical products (TEMPs) or other collagen-based biomaterials, for product quality control. This method also can be used for qualitative analysis and quantitative detection of bovine-specific and/or type I-specific collagen in the samples mixed with other animal sources and/or other type collagen. This document does not exclude other possible methods for quantifying type I collagen, such as hydroxyproline quantification, that can evaluate the total amount of collagen regardless of type. NOTE 1 The collagen has been known there are greater than 28 types and with the different property in each one. This document focuses on the quantification of marker peptide of purified bovine type I collagen. Type I collagen isolated from skin, tendon, bone, etc., can contain other types of collagen, for example, type III and type V. And type I collagen can be sourced from bovine, swine, etc. For quantification of other types of collagens or type I collagen sourced from other species of animals can use this document as a template, but need to design collagen type-specific or/and animal species-specific characteristic peptides for LC-MS/MS method, as well as optimize the determination conditions. NOTE 2 For quantification of the collagen marker peptide of scaffold which combined with other materials, or type I collagen contained in ECM materials of tissues or type I collagen-based regenerative tissues, can refer to this document, but need to isolate or/and purify the type I collagen with a reasonable and verified method at first (9,10), and then quantify it by referring to the method provided in this document.