Boiling point:488.7ºC at 760 mmHg
Molecular formula:C3H7MgO6P
Molecular weight:194.363
Flash point:249.4ºC
Accurate mass:193.983063
PSA:122.69000
1.Periodic acid method
Principle: Periodic acid oxidation method is a common method for the detection of glycerol content at present, the use of periodic acid and glycerol reoxidation reaction, the remaining periodic acid and the generated iodic acid reaction with potassium iodide reaction to produce iodine, and then titration with sodium thiosulfate, according to the amount of sodium thiosulfate consumed to calculate the content of glycerol. 2. The fermentation liquid 1{{10}}m1 is dissolved in a 100mL small beaker with a small amount of water, then transferred to a 100mL volumetric bottle, diluted with water to the line, shaken well, and left to stand. Use a pipet to accurately remove 10mL of the above solution into the iodine measuring bottle, add 20m sodium periodate solution, mix evenly, stand in the dark for 40min away from light, then add 15mL potassium iodide solution and 20% hydrochloric acid solution, adjust the pH value between 0.8 and 1.0, and then titrate with sodium thiosulfate standard solution. Near the end, 2 starch indicator solution was added and titrated until the blue color of the solution disappeared. At the same time do sample blank.
2, enzyme colorimetry
1. Principle: It is a enzymatic method for the determination of glycerol established by the specific catalytic reaction of enzymes, which refers to the conversion of glycerol into 3-phosphate glycerol under the action of glycerol kinase, which is catalyzed by phosphoglycerol oxidase to generate dilight acetone phosphate and hydrogen peroxide. Then hydrogen peroxide and 4-aminopyrine, 4-chlorophenol react under the catalysis of peroxidase to produce purple and blue color, can have a characteristic absorption peak at about 500m of the imine, through the color depth, that is, absorbance change determination of the generated H,O, glycerin content and H0 is proportional to the generated, so that the colorimetric method can be used to determine the content of glycerin.
2. Operation Procedure Wavelength :500nm (480~520nm) Reaction temperature :37C Colometric cup Light diameter :1cm Take a certain amount of R1(see R2 bottle tag) and add it to a bottle of R2, which is the working liquid after dissolution, and the working liquid is pre-insulated to the test temperature
3, high performance liquid chromatography
Principle: The principle of high performance liquid chromatography (HPLC) is based on the liquid as the mobile phase, the use of high pressure infusion system, the single solvent with different polarity or different proportions of mixed solvents, buffers and other mobile phases are pumped into the column equipped with a fixed phase, the column is used to separate the test mixture first, and then detect, so as to achieve the analysis of the sample. Therefore, the accuracy and precision of the measurement are high, and it has become a commonly used method for detecting biochemical molecules. 2. Procedure
(1) First, the fermentation solution was diluted 2.5 times with water, and then HPO was added to adjust H to 55 at a rotational speed of 8000r/min and a temperature of 4C for 10min, and then filtered by microporous filtration membrane for HPLC analysis :(2) The glycerol standard series solution was carried out by HPLC under the best chromatographic conditions Analysis, using peak area external standard method to quantify the glycerol standard curve:
(3) After sample pretreatment, 5 µl was injected, the peak area was recorded, substituted into the linear equation, and its measured value was calculated. The calculated value is multiplied by the dilution times to obtain the glycerol content in the fermentation solution.
4,Gas chromatography
1. Principle: Gas chromatography is a method for separating and analyzing compounds in complex samples. The principle is that a certain amount of gas or liquid analyte is injected into the injection mouth at one end of the column, driven by the carrier gas through the column, and the molecules of the analyte will be adsorbed by the column wall or the packing in the column. Because different samples have different physical and chemical properties, and have different interactions with specific stationary phases, each type of molecule has its own rate of passage, so that various components of the analyte will reach the end of the column at different times, and thus get separated. When the compounds flow out of the end of the column, they are detected by the detector, producing the corresponding signal, which is converted into an electrical output, thereby determining the time order in which each component reaches the end of the column and the amount of each component.
