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[Content Determination] Preparation of Polysaccharide Reference Substance Solution:Take an appropriate amount of anhydrous glucose reference substance, accurately weigh it, and add water to make a solution containing 0.12 mg per 1ml.
Preparation of the Standard Curve: Precisely measure 0.2ml , 0.4ml , 0.6ml , 0.8ml , 1.0ml , 1.2ml of the reference solution, Put into 10ml graduated test tube with stopper , add water to each to 2.0ml, and add 6ml of the anthrone-sulfuric acid solution (weigh the anthranone 0.1g precisely, dissolve with 100ml sulfuric acid and shake well) quickly and accurately, shake immediately, place it for 15 minutes, immediately place it in an ice bath to cool for 15 minutes and take it out. With the corresponding reagent as a blank, according to the ultraviolet-visible spectrophotometry, measure the absorbance at a wavelength of 625nm, draw a standard curve with absorbance as the ordinate and concentration as the abscissa.
Preparation of Test Solution: Take about 2g of ganoderma lucidum powder, accurately weigh it, put it in a round bottom flask, add 60ml of water and left it for 1 hour, heat the solid flow for 4 hours, filter while hot, and wash the filter and filter residue with a small amount of hot water. Put the filter residue and filter paper in a flask, add 60ml of water, heat to reflux for 3 hours, filter while hot, combine the filtrate, put it on a water bath and steam to dry, dissolve the residue with 5ml of water, slowly add 75ml of ethanol while stirring, shake well, leave it at 4 ° C for 12 hours, centrifuge, discard the supernatant, dissolve the precipitate with hot water and transfer to a 50ml measuring flask, let it cool, add water to the mark, shake well. Take an appropriate amount of the solution, centrifuge, accurately measure the supernatant 3ml and put it In a 25ml measuring flask, add water to the mark, shake well, and then it is ready.
Detection Method: Take 2ml the test sample solution accurately and put it in a 10ml graduated test tube with stopper. According to the method under the preparation of the standard curve , from "Quickly and precisely add 6ml of onion ketone sulfate solution", operate in the same way to determine the absorbance. Read the content of anhydrous glucose in the test solution accordingly to the standard curve, calculate it and get it.
This product is calculated as a dry product, and the content of Ganoderma lucidum polysaccharide is calculated as anhydrous glucose (C6H12O6), not less than 0.90%.
Triterpenes and Sterols
Preparation of Reference Substance Solution: Take an appropriate amount of oleanolic acid reference substance, accurately weigh it, and add methanol to make a solution containing 0.2 mg per 1 ml.
The Preparation Precision of the Standard Curve: Take 0.1ml, 0.2ml, 0.3ml, 0.4ml, 0.5ml of the reference solution, put them in 15ml Graduated test tube with stopper, dry it and let it cool.Precisely add 0.2ml of freshly prepared vanillin Glacial acetic acid solution (precisely weigh 0.5g of vanillin and add glacial acetic acid to dissolve it into 10m to get it) and 0. 8ml of perchloric acid , shake well. Heat it in a water bath at 70°C for 15 minutes, immediately place it in an ice bath to cool for 5 minutes, then take it out, add 4ml of ethyl acetate precisely and shake it well. With the corresponding reagent as a blank, according to ultraviolet-visible spectrophotometry, measured at a wavelength of 546nm for absorbance, draw a standard curve with absorbance as the ordinate and concentration as the abscissa.
Preparation of Test Solution: Take about 2g of ganoderma lucidum powder, accurately weigh it, put it in a stoppered conical flask, add 50ml of ethanol, ultrasonic treatment (at power 140W, frequency 42kHz) for 45 minutes, filter, and put the filtrate in a 100ml measuring flask. Use proper amount of ethanol to wash the filter and filter residue in batches. Put all the lotion into the same measuring bottle, add ethanol to the mark, shake well and get it.
Detection Method: Precisely measure 0.2ml of the test solution and put it into a 15ml Graduated test tube with stopper. According to the method under the preparation of the standard curve, Starting from "Volatilization", operate in the same way to measure the absorbance.. Read the content of anhydrous glucose in the test solution accordingly to the standard curve, calculate it and get it.
This product is calculated as a dry product and the contents of triterpenoids and sterols are measured by oleanolic acid (C30H48O3) and shall not be less than 0. 50%.
