Hydrogen sulfide content was quantified colorimetrically tissue detection kit product specification (Chinese version)

The main purpose

Tissue hydrogen sulfide content colorimetric quantitative detection reagent is a change in the absorption peak caused by the reaction of basic picric acid with creatinine to produce a blue product, that is, the use of colorimetry to determine the hydrogen sulfide content of the sample. And the classic technical method. This technology has been proven through careful modification of the methylene blue method and successful experiments. It is suitable for the detection of hydrogen sulfide levels in various human and animal tissue lysis samples. The product is strictly sterile, ready to use, simple in operation and stable in performance.

technical background

Hydrogen sulfide (H2S) is a naturally occurring, corrosive and flammable, unstable, oxidizable, colorless gas that is a desulfuration metabolite of cysteine ​​and is the third physiology. A gas messenger molecule or a gasophore molecule exists in the brain, heart, lung, liver, small intestine, pancreas, and cavernosum tissues, in which brain tissue content is low and kidney tissue content is high. . Nervous system cystathionine -β - synthase (Cystathionine-β-synthase; CBS ) , and cardiovascular system cystathionine - γ- lyase (cystathionine γ -lyase; CGL) is involved in the generation of endogenous hydrogen sulfide main members. Once the body is deprived of oxygen, the hydrogen sulfide content increases. Hydrogen sulfide is intended to regulate the level of sulfur-containing amino acids in tissues, and has physiological functions and pathophysiological regulation of the cardiovascular system, such as induction of vascular smooth muscle relaxation, vasodilation, decreased blood pressure, and pro-inflammatory effects. At the same time, hydrogen sulfide has a toxic effect, which causes rapid convulsions of the olfactory nerve, inhibits cytochrome c oxidase, increases the opening of the mitochondrial membrane channel, and causes excessive relaxation of the heart, small intestine, reproductive system, urinary system and respiratory smooth muscle, resulting in organ damage, Hypotension, bradycardia and other symptoms. In the natural environment, hydrogen sulfide is the main intermediate of the sulfur cycle, including mineralization or decomposition of organic sulfides, assimilation of microorganisms, and redox. As a chemical asphyxiant gas, hydrogen sulfide is toxic to fish and other aquatic animals and is also a marker of bacterial contamination. Zinc acetate is used to capture hydrogen sulfide under alkaline conditions to form a stable zinc sulfide precipitate; under acidic conditions, dissolved zinc sulfide and catalyst p-aminodimethylaniline (N, N-dimethyl-p- Phenylenediamine reacts with ferric chloride to produce blue methylene blue, which is quantitatively analyzed for its hydrogen sulfide content by its peak absorption peak (wavelength at 670 nm).

product content

Reagent A Reagent B Reagent C Reagent C Reagent D Reagent E Reagent E Reagent F ml Product Specification 1 Recipient

storage method

The reagent solution (Reagent D) and GENMED standard solution (Reagent F) are stored in a refrigerator at -20 ° C, and the rest are stored in a refrigerator at 4 ° C; the reaction solution (Reagent D) is protected from light; the reagent is corrosive, and the operation is safe; Effectively guaranteed in June

User-supplied

1.5 ml centrifuge tube: container for sample handling
15 ml conical centrifuge tube: container for sample handling
DOUNCE homogenizer: micro-tabletop centrifuge for lysing tissue cells: for sample manipulation cuvettes: container spectrophotometer for colorimetric: for colorimetric analysis

Experimental procedure

Sample preparation

• Surgical removal of animal tissue and weighing to determine 200 mg tissue weight
• (selection step) into a pre-cooled 15 ml conical tube
• (Selection step) Add 1x cleaning solution (Reagent A)
• Immediately shredding tissue with a blade
• Place a pre-cooled 15 ml conical tube
• Add pre-cooled xx ml of cleaning solution (Reagent A)
• Vortex for 5 seconds, mix thoroughly
• Immediately place in a pre-cooled DOUNCE homogenizer and homogenize the tissue with a homogenized rod in an ice bath (approximately 40 to 80)
• Transfer all tissue homogenates into a 1.5 ml centrifuge tube
• Place in a 4°C benchtop centrifuge for 10 minutes at a speed of 10,000g
• Pipette 500 μl into a new 1.5 ml centrifuge tube
• Add xx μl of Stabilizer (Reagent B ) and mix
• Place in the ice tank for testing (Note: must be used within 1 week)

Standard sample preparation

• Transfer xx microliters of standard solution (Reagent F) to a new 1.5 ml centrifuge tube
• Add xx μl of Stabilizer (Reagent B ) and mix
• Placed in the ice trough to be tested

Preparation for measurement

• Prepare the sample to be tested and place it in the ice trough
• Set the spectrophotometer: wavelength 670nm, and set zero

Sample determination

• L xx to interference pipetted liquid (Reagent C) to said sample to be tested or standard sample in the tube, mix
• Add xx microliters of reaction solution ( Reagent D )
• Add xx microliters of Reagent E and mix
• Incubate for 20 minutes at room temperature
• Place in a mini tabletop centrifuge for 3 minutes at a speed of 5000g
• Pipette 1 ml of supernatant into cuvette
• Immediately put into the spectrophotometer to obtain the sample to be tested and the standard sample reading

Fourth, calculate the sample concentration

Converted to the actual hydrogen sulfide content of the tissue

Precautions

• This product is 20 operations, including standard samples
• The detection range of this product is 0.1 mg / liter (10 μmol / liter) to 8.5 mg / liter (250 μmol / liter)
• Wear gloves when handling
• Reagents are corrosive, pay attention to operational safety
• Standard sample determination is only 1 time during system operation
• Precipitation of the reagent can be removed after centrifugation
• The measured value changes from low to high
• Users can detect at any wavelength in the 664nm to 670nm wavelength range
• After the colorimetric determination, the cuvette must be thoroughly cleaned.
• If the concentration of the sample to be tested is too high or too low, the sample amount can be adjusted.
• Iodine (iodine) in the sample is greater than 2 mg / liter, thiosulfate or sulfite greater than 10 mg / liter, ferrocyanide and other possible interference detection
• Hydrogen sulfide concentration conversion: 1 mg / liter = 29.4 micromol / liter
• The company provides a series of biochemical detection reagent products

Quality Standard

• This product has been certified to be stable.
• This product has been identified and sensitive