Pretreatment method for quantitative detection of undenatured type ii collagen in collagen product or cartilage, and application
AssigneeBeijing Semnl Biotechnology Co., Ltd.
InventorsLige LIU, Zheng YAN, Aiqing LIU, Haiyan WANG, Guoqing ZOU
Life of this patent
Estimating the term…
Abstract
A pretreatment method for detection of an undenatured type II collagen in a cartilage collagen product or a cartilage is proposed. The method can separate an undenatured type II collagen from a complex environment system including proteoglycan, a hydrolyzed collagen, open-chain denatured collagen and the like, and present a dissolved state, thereby facilitating subsequent further qualitative and quantitative detection.
Claims21
A pre-treatment method for detection of non-denatured type II collagen in a collagen product or cartilage raw material, comprising:
- (1) adding a buffer solution containing a neutral salt or guanidine hydrochloride to a sample of the collagen product or cartilage raw material to be tested, centrifuging, and obtaining a precipitate;
- (2) resuspending the precipitate obtained in (1) with an acid, accelerating the swelling by ultrasonication, and centrifuging to obtain a supernatant;
- (3) adding pepsin to the supernatant obtained in (2) for enzymatic digestion;
- and
- (4) adding elastase to the enzymatic digest obtained in (3) for enzymatic digestion, centrifuging, and retaining the supernatant as the solution to be tested.
A method for detecting non-denatured type II collagen in a collagen product or cartilage raw material, comprising:
- (1) adding a buffer solution containing a neutral salt or guanidine hydrochloride to a sample of the collagen product or cartilage raw material to be tested, centrifuging, and obtaining a precipitate;
- (2) resuspending the precipitate obtained in (1) with an acid, accelerating the swelling by ultrasonication, and centrifuging to obtain a supernatant;
- (3) adding pepsin to the supernatant obtained in (2) for enzymatic digestion;
- (4) adding elastase to the enzymatic digest obtained in (3) for enzymatic digestion, centrifuging, and retaining the supernatant as the solution to be tested;
- and
- (5) performing qualitative and/or quantitative analysis of collagen on the solution to be tested obtained in (4).
A kit for the detection of non-denatured type II collagen in a collagen product or cartilage raw material, wherein said kit comprises a buffer solution containing a neutral salt or guanidine hydrochloride, a pepsin solution, and an elastase solution.
(canceled)
Dependencies are inferred from the claim text.