The product is a neutral polysaccharide produced in the process of culturing Aureobasidium pullulans. 3 glucose through α-1,4 glycosidic bonds to form maltotriose, and then repeated bonding through α-1,6 glycosidic bonds to make the structure of the pullulan.
Properties
the product is white to off-white non-crystalline powder, odorless and tasteless.
Solubility
the product is easily soluble in water and almost insoluble in ethanol, ether and glycerin.
Identification
- Take about 10g of the product, add water of 100ml, stir constantly to dissolve the product, and it should be a viscous solution.
- Take 2 tubes, add 10ml of above viscous solution to each tube, add pullulanase solution of 0.1ml (10 units/ml) to the first tube, add water of 0.1ml to the second tube, shake well and leave them under the room temperature for 20 minutes. The viscosity of the solution in the first tube should be significantly lower than that in the second tube.
- Take 10 ml of the solution of step 1, dilute it to 50 ml with water, shake well, and then, take 10 ml, add 2 ml of polyethylene glycol 600, and a white precipitate is produced immediately.
Viscosity
take the product, using vacuum drying method at 105 °C for 2.5 hours. Accurately weigh 10.0g, add water to dissolve the product and dilute the solution to 100g. According to the rule (Chinese Pharmacopoeia 2015 General Rules 0633 first method), at 30 ± 1 ° C (the capillary inner diameter of 2.0 mm), the kinematic viscosity should be 50-180 mm2 /s.
PH
take 1.0g of this product, add 10mL of distilled water cooled after boiling 10mL for dissolution. The PH value should be 4.5~6.5 at 25 °C.
Monosaccharides and oligosaccharides
test based on UV-visible spectrophotometry
Steps1
Take the product, vacuum drying at 90 °C for 6 hours. Accurately weigh 0.80 g, add water for dissolution and dilute it to 100 ml, then comes out the stock solution. Precisely measure 1 ml of stock solution, add 0.1 ml of saturated potassium chloride solution and 3 ml of methanol, then mix them up, and centrifuge. Take the liquid supernatant as a test solution for monosaccharides and oligosaccharides. In addition, accurately take 1 ml of the stock solution, r and diluted it to 50 ml with water, then come out a total sugar solution. Separately measure 0.2 ml of the monosaccharide and oligosaccharide test solution, total sugar solution and water. slowly add them to 5ml of 75% sulfuric acid aqueous solution which is cooled with cold water and containing 0.2% fluorenone, mix immediately, and put the solution in a water bath of 90°C for 10 minutes, then immediately cool the temperature of the solution down to room temperature, and shake well. Measure the absorbances At, As, Aw at a wavelength of 620 nm. Calculate the content of monosaccharides and oligosaccharides according to the following formula, and it should not exceed 10.0%.
Content% = (At-Aw) / (As-Aw) * 8.2
In the formula
At = the absorbance of the test solution of monosaccharides and oligosaccharides
As = the absorbance of the total sugar solution
Aw = the absorbance of water
Steps2
Standard solution: weigh 0.2g of glucose, accurate to 0.001g. add water for dissolution and dilute it to 1 L. take 0.2ml of the solution and add it to 5ml of anthrone solution, mix well.
Blank solution: add 0.2 ml of water to 5 ml of anthrone solution, and mix well.
Sample stock solution: weigh 0.8g sample, accurate to 0.001g, add water for dissolution and dilute it to 100ml.
Sample solution: Measure 1 ml of the sample stock solution, place it in a centrifuge tube, add 0.1 ml of saturated potassium chloride solution and 3 ml of methanol, vigorously mix for 20S, and centrifuge at a speed of 11000r/min for 10min. Take 0.2 ml centrifuged liquid supernatant and add it to 5 ml of an anthrone solution, and mix them well.
Take the sample solution, the standard solution and the blank solution, and keep them warm in a 90 ° C water bath for 15 min. Use a spectrophotometer to test absorbance of these solutions at 620 nm. The content of monosaccharides, disaccharides and oligosaccharides is calculated by the mass fraction of glucose W1, and the value is expressed in %. The content of monosaccharides, disaccharides and oligosaccharides should not exceed 10%, and is calculated according to the following formula:
Content % = ((At - Ab) * 0.41 * m1) / ((As - Ab) * m0)
In the formula
At = the absorbance of the sample solution
As = the absorbance of the standard solution
Ab = the absorbance of the blank solution
m1 = glucose quality
m0 = sample quality
Nitrogen
take about 3g of the product, the amount of sulfuric acid used in digestion and decomposition is 12ml, and add 40ml of 40% sodium hydroxide solution, distill the solution, test the dried product according to nitrogen testing method, and the nitrogen content should be less than 0.05%.
Loss on drying
take the product, vacuum drying at 90 °C for 6 hours. The weight loss should not exceed 6.0%.
Residue on ignition
Take 2.0g of the product, check it accordingly, and the residual residue should not exceed 0.3%.
Heavy metal
take the residue after the ignition residue test, check according to the heavy metal inspection method, and the heavy metal should not exceed 20 parts per million.
Microbiological limit
Take the product, test according to the microbiological limit inspection of non-sterile products: microbiological counting method and non-sterile product microbial limit inspection: control bacteria inspection method. The total number of aerobic bacteria per 1g test sample should not exceed 1000cfu, the total number of molds and yeasts should not exceed 100 cfu, and Escherichia coli should be absent;
Category
pharmaceutical excipients, binders and fillers.
Storage
protect the product from light and keep it sealed.
