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Method for quantitatively detecting coagulant activity of bean phytohemagglutinin

A quantitative detection method, plant lectin technology, applied in the field of plant component detection, can solve the problems of large error in results, inconsistent methods, large randomness, etc., and achieve the effect of accurate results, simple steps, and small error

Active Publication Date: 2014-02-05
JIANGHAN UNIVERSITY
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Problems solved by technology

These methods have the following disadvantages: (1) only qualitative or semi-quantitative determinations can be made; (2) there are different methods for preparing standard red blood cell suspensions, which are mostly expressed by volume percentage, and the reproducibility is poor; (3) the randomness of determining the reading end time The results are relatively large, some have visual observation results, the subjectivity is large, the result error is also large, and the accuracy is low; (4) There are various measurement methods, and the measured results are very different, so it is impossible to compare

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  • Method for quantitatively detecting coagulant activity of bean phytohemagglutinin

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Embodiment Construction

[0032] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0033] (1) Preparation of standard red blood cell suspension and determination of standard curve:

[0034] 1. Collect 2ml of fresh rabbit blood by heart blood collection, put it in 300ul of 8% sodium citrate anticoagulant, mix it with 8ml of Alsever's solution, and store it in a 4°C refrigerator for later use (within 2 weeks, the red blood cells are normal, more than 2 weeks, red blood cells appear hemolysis).

[0035] 2. Take 2.5ml of fresh rabbit blood preserved in Alsever’s solution, slowly mix it with normal saline at a ratio of 5:8, centrifuge at 800-1000rpm for 10 minutes, and collect the red blood cell pellet.

[0036] 3. After repeating the above steps to wash the red blood cells five times, the color of the supernatant is completely transparent, and then according to the volume of the red blood cell sedimentation, make a 20% red bloo...

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Abstract

The invention discloses a method for quantitatively detecting the coagulant activity of bean phytohemagglutinin and belongs to the technical field of plant component detection. According to the method, a red cell suspension for detection is subjected to cell quantification; meanwhile, a coagulant activity concentration standard curve is drawn by using the bean phytohemagglutinin with the known concentration as a standard substance, and the coagulant activity concentration of the bean phytohemagglutinin is further calculated, so that the coagulant activity of the bean phytohemagglutinin is quantified.

Description

technical field [0001] The invention belongs to the technical field of plant component detection, and in particular relates to a quantitative detection method for coagulation activity of kidney bean lectin. Background technique [0002] The lectins contained in kidney beans are one of the poisonous substances that cause food poisoning in kidney beans, especially the pods of fresh kidney beans, which often cause poisoning incidents due to improper cooking. Kidney bean poisoning is multifactorial, and the most important factor is that the lectin in kidney bean can recognize and specifically combine with the specific glycosyl groups on the surface of red blood cells to form many crossed "bridges" on the surface of red blood cells, causing the red blood cells to agglutinate, which in turn causes The overall response of the body. The content of lectin in different bean varieties is not the same, so the hemagglutination method can be used to detect the lectin activity in the bean...

Claims

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Application Information

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IPC IPC(8): G01N21/33G01N1/28
Inventor 李佳楠陈禅友潘磊
Owner JIANGHAN UNIVERSITY
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