Bilberry Extract Method of Analysis

- Jan 22, 2019-


[Source] This product is a fresh fruit extracted from Vaccinii vitis-idaeae of Ericaceae.

[Properties] This product is purple-brown powder.

[Identification] (1) Take appropriate amount of this product, add dilute hydrochloric acid 10 ml to dissolve, add ammonia test solution to adjust PH9.0, the solution from red to blue.

Error reporting (2) Dissolve the product with diluted hydrochloric acid to make a solution containing 10 UG per ml of sample. Photometric method (Appendix V A of 2010 edition of Chinese Pharmacopoeia) test shows that the absorption is maximum in the wavelength range of 530-550 nm.

[Inspection]

Particle size: Take 10g of this product, pass 80 mesh screen, should pass all.

Moisture: No more than 5%.

Ash content: no more than 1.0% (Appendix IX K, 2010 edition of Chinese Pharmacopoeia).

Heavy metals: no more than 10 parts per million (Act II of Appendix IX E, 2010 edition of Chinese Pharmacopoeia).

Arsenic salt: not more than two parts per million (the first method under Appendix IX F of the 2010 edition of Chinese Pharmacopoeia).

[Content determination] Anthocyanins

Method 1: Precision weighing 10mg, adding 2% methanol hydrochloride solution 60ml, heating reflux for 80 minutes, cooling in water bath and adding 2% methanol hydrochloride solution into 100ml volumetric flask, precisely taking 2ml of the solution, adding 2% methanol hydrochloride solution diluted to 25ml, shaking, and measuring the absorbance at 530nm according to the spectrophotometric test (Appendix V A of 2010 edition of Chinese Pharmacopoeia). At the same time, blanks are made and the content is calculated according to the following formula:

image

A: Absorbance  E:Percentage absorption coefficient of cyanidinimage =1020

M: Sample weightg

This product contains total anthocyanin according to the dried product, and cyanidin should not be less than 25.0%.

Method 2: The method was determined by high performance liquid chromatography.

Chromatographic conditions and system adaptability test: Zorbax SB C18 (4.6 *250mm) 5 micron was used as filler; detection wavelength was 525 nm; theoretical plate number should not be less than 5000 according to cyanidin-3-glucoside peak. With A:10% formic acid and B:methanolic acid as mobile phase, the column temperature was maintained at 40 +0.5 degrees C and the flow rate was 1.0 ml per minute. The elution was carried out according to the following table conditions:

Timemins

A

B

0-20

95-40

5-60

20-25

40-0

60-100

25-30

0

100

 

Preparation of reference solution: Precise weighing of cypermethrin-3-O-glucoside reference 5 mg, placed in 25 ml flask, add 0.1% methanol hydrochloride solution to dissolve into the scale, shake well, then get.

Preparation of sample solution: Precision weighing 100mg of this product, placed in 25ml measuring bottle, add 0.1% methanol hydrochloride solution to dissolve to the scale, shake well, then get.

Determination method: Precision absorption of reference solution and sample solution of 20 ml, injection of liquid chromatography, according to the following calculation.

For Example:

A: Total anthocyanins

B. Reference substance concentration

C. Sample solution volume

D. Sample weight

E. Percentage of reference peak area

image 

This product is calculated according to the dried product, and the total anthocyanin content is calculated by cyanidin-3-O-glucoside, which should not be less than 35.0%.

[Specification] Contains 35% anthocyanin.

[Storage] Sealed, moisture-proof.


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