A method for detecting the content of main components in heparan sulfate finished products
A technology of heparan sulfate and dermatan sulfate, which is applied in the field of chemical detection, can solve the problems that the content of heparan sulfate cannot be directly measured and is difficult to separate, and achieves the effects of high accuracy, good repeatability and simple detection method
Active Publication Date: 2022-04-26
深圳市格利特生物科技有限公司
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[0002] Heparan sulfate (Heparin Sulfate, referred to as HS) is a kind of glycosaminoglycan widely present in animal tissue matrix and cell membrane. Uronic acid or iduronic acid is connected to form a disaccharide unit. In addition to the main component of heparan sulfate, the finished product of heparan sulfate is also accompanied by a small amount of impurities such as dermatan sulfate (Dermatan Sulfate, DS), due to heparan sulfate and The properties of dermatan sulfate are similar, so it is difficult to separate them directly by liquid phase methods. Even under the chromatographic conditions of the United States Pharmacopoeia Heparin Sodium Chromatographic Identification Instrument, heparan sulfate and dermatan sulfate peak at the same peak time, so it is impossible to directly determine sulfuric acid. Heparan content
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[0035] The claims of the present invention will be further described in detail below in conjunction with specific embodiments.
[0036] It should be noted that all the contents detected in this application are mass percentages.
[0037] 1. Detection of DS mass percentage content
[0038] Principle: Use chondroitin sulfate enzyme B (DS enzyme) as a tool to specifically degrade dermatan sulfate in heparan sulfate into fragments mainly composed of DS disaccharides, and use high performance liquid chromatography to detect disaccharides in samples DS The peak area was used to calculate the content of dermatan sulfate.
[0039] 1.1. DS disaccharide of the sample
[0040] Take 100mg of a certain batch of heparan sulfate finished product, dissolve it in 50ml Tris-HCl buffer solution (50mM, containing 10mM CaCl2, pH7.0), and prepare 2mg / ml heparan sulfate mother solution, take 2μl of the mother solution into 50mM Tris-HCl buffer solution Dilute to 0.5ml, place in a 1.5ml centrifuge ...
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The invention discloses a method for detecting the content of main components in heparan sulfate finished products, and aims to provide a detection method for the content of main components in heparan sulfate finished products with simple detection method, strong specificity, high accuracy and good repeatability ; Its scheme is such: 1) adopt DS enzyme to degrade the dermatan sulfate in the heparan sulfate finished product into the solution of DS disaccharide; Samples were injected into the high performance liquid chromatography to detect the disaccharide peak area of each gradient of DS, and the DS working curve was drawn; 3) the high performance liquid chromatography method was used to detect the DS disaccharide in the sample solution after step 1) degradation, and the DS content was calculated ; 4) Detecting the peak area of DS+HS with high performance liquid chromatography, and calculating the content of DS+HS; 5) calculating the content of HS; belonging to the technical field of chemical detection.
Description
technical field [0001] The invention relates to a method for detecting the content of main components in heparan sulfate finished products, and belongs to the technical field of chemical detection. Background technique [0002] Heparan sulfate (Heparin Sulfate, referred to as HS) is a kind of glycosaminoglycan widely present in animal tissue matrix and cell membrane. Uronic acid or iduronic acid is connected to form a disaccharide unit. In addition to the main component of heparan sulfate, the finished product of heparan sulfate is also accompanied by a small amount of impurities such as dermatan sulfate (Dermatan Sulfate, DS), due to heparan sulfate and The properties of dermatan sulfate are similar, so it is difficult to separate them directly by liquid phase methods. Even under the chromatographic conditions of the United States Pharmacopoeia Heparin Sodium Chromatographic Identification Instrument, heparan sulfate and dermatan sulfate peak at the same peak time, so it is...
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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 黄洪陈海苑
Owner 深圳市格利特生物科技有限公司