An agent for quantitative detection of fructose in seminal plasma of indole method, kit, and method for detecting fructose in seminal plasma
A sugar indole method, quantitative detection technology, applied in measuring devices, color/spectral characteristic measurement, instruments, etc., can solve problems such as easy corrosion and damage to instruments, difficult to seal, easy to pollute the environment, etc., to improve detection accuracy, environmental protection As a result, the effect of reducing interference
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Embodiment 1
[0022] Embodiment 1, a kit for quantitative detection of seminal berry fructose indole method, comprising:
[0023] a) Protein-removing solution: 1-3% ZnSO 4 ·7H 2 O aqueous solution;
[0024] b) Alkaline solution: 0.02-0.5mM NaOH aqueous solution;
[0025] c) Chromogenic solution: 2×10 -3 ~10×10 -3 % indole, 0.1-0.3% benzoic acid mixed aqueous solution;
[0026] d) 37% concentrated hydrochloric acid;
[0027] e) Protective agent: Tween-80, glycerin mixture, glycerin accounts for 30-70% of the total volume;
[0028] f) Fructose standard solution: a mixed aqueous solution of 1.12-3.36mM fructose and 0.1-0.3% benzoic acid;
[0029] g) an Eppendorf reaction tube whose inner wall is coated with a methyl silicone oil isolation membrane.
[0030] In this example, the alkaline solution can also be other alkaline solutions such as sodium carbonate solution; the chromogenic solution can also be a solution that can react with fructose to generate a colored product such as resorc...
Embodiment 2
[0031] Embodiment 2, a detailed production method of the seminal berry fructose indole method quantitative detection kit, comprising the following steps:
[0032] a) Preparation of deproteinized solution
[0033] i) Take ZnSO 4 ·7H 2 O1 to 3 grams, put in a beaker and add appropriate amount of deionized water to dissolve;
[0034] ii) Dilute to 100 ml with deionized water, shake and filter;
[0035] b) preparing an alkaline solution;
[0036] i) Put 100-120 grams of AR grade NaOH in a beaker, add 200 ml of deionized water, stir the beaker in cold water, pour it into a polyethylene plastic bottle after dissolving, and let it stand at room temperature for several days.
[0037] ii) Take an appropriate amount of concentrated NaOH supernatant, put it into a 500 ml volumetric flask, and add deionized water to the desired concentration.
[0038] c) preparing a chromogenic solution;
[0039] i) Take 1 to 3 grams of benzoic acid, add 900 ml of deionized water, shake and dissolve...
Embodiment 3
[0049] Embodiment three, a whole process of the method for the quantitative detection of seminal berry fructose using the seminal berry fructose indole method quantitative detection kit prepared in Example 2, the specific steps are:
[0050] a) The semen sample to be tested is centrifuged at 1000 g for 10 minutes, and the seminal plasma is poured out.
[0051] b) Take 20ul of seminal plasma, add 200ul of diluent, 50ul of deproteinized solution, and 50ul of alkaline solution, mix, place at room temperature for 15 minutes, centrifuge at 2000g for 20 minutes or 8000g for 5 minutes, take 50ul of supernatant, and put it into the inner wall to coat nail The Eppendorf tube of silicone oil-based isolation film is used as the tube to be tested;
[0052] c) Dilute the 2.24mM fructose standard solution down to 1.12mM, 0.56mM, 0.28mM, 0.14mM, use deionized water as the zero fructose standard solution, take 50ul each and put it into the inner wall to coat the methyl silicone oil isolation ...
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