Standards Development

Scope

This standard 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 standard 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.

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., may contain other types of collagen, for example, type III and type V. And type I collagen may source from bovine, swine, etc. For quantification of other types of collagens or type I collagen sourced from other species animals may use this document as a template, but need 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, may refer to this document, but need to isolate or/and purify the type I collagen with a reasonable and verified method at first (Pataridis S, 2008; Pataridis S, 2009), and then quantify it by referring to the method provided in this document.

NOTE 3 If the experimental conditions (e.g. no equipment) are not available for applying the method provided in this document, the other methods can be chosen, e.g. using hydroxyproline detection as an indirect quantitative method for total collagen content determination, or using enzyme immunoassays with antibodies specific to each collagen type for qualitative analysis and relative quantitative determination. The disadvantages of these methods are described in the introduction of this document.