Determination method of Chlorogenic acid and Luteoloside content in Honeysuckle Flower Extract
[Content determination] Determined according to HPLC determination.
Chromatographic conditions and system suitability test:
Use ODS as filler; use acetonitrile-0.4% phosphoric acid solution (13:87) as mobile phase; detection wavelength as 327nm. The number of theoretical plates should not be less than 1,000 based on the peak of chlorogenic acid.
Preparation of reference substance solution:
Take an appropriate amount of chlorogenic acid reference substance, accurately weigh it, place it in a brown measuring flask, add 50% methanol to make a solution containing 40 µg per 1 ml, and it will be obtained (stored below 10°C).
The preparation of the test solution:
Take about 0.5g of this product powder (passed through No. 4 sieve), accurately weigh it, place it in a stoppered conical flask, add 50% methanol 50ml accurately, weigh it, and ultrasonically treat it (power 250W, frequency 35kHz) for 30 minutes. Cool, weigh it again, use 50% methanol to make up the lost weight, shake well, filter, accurately measure 5ml of the filtrate, place it in a 25ml brown measuring flask, add 50% methanol to the mark, shake it, and get it.
Detection method:
Precisely draw 5-10 µl each of the reference solution and the test solution,inject it into the HPLC instrument for determination.
This product is calculated as a dry product and contains chlorogenic acid (C16H18O9) not less than 1.5%.
Determination of Luteoloside according to HPLC.
Chromatographic conditions and system suitability test:
Use phenylsilane-bonded silica gel as a filler (Agilent ZORBAX SB-phenyl 4.6mm×250mm, 5μm), use acetonitrile as mobile phase A, 0.5% glacial acetic acid solution as mobile phase B, and perform gradients as specified in the following table Elution; detection wavelength is 350nm. The number of theoretical plates should not be less than 20000 calculated based on the luteolin peak.
Time (minutes) Mobile phase A (%) Mobile phase B (%)
0〜15 10→20 90→80
15 〜30 20 80
30 〜40 20→30 80→70
Preparation of reference substance solution:
Take an appropriate amount of luteolin reference substance, accurately weigh it, and add 70% ethanol to make a solution containing 40 µg per 1 ml.
Preparation of the test solution:
Take about 2g of the powder (passed through the No. 4 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50ml of 70% ethanol, weigh it, and ultrasonically treat it (power 250W, Frequency 3 5 kHz) for 1 hour, let it cool, then weigh it, use 70% ethanol to make up the lost weight, shake well, and filter. Accurately measure 10ml of the continued filtrate, recover the solvent to dryness, dissolve the residue with 70% ethanol, transfer to a 5ml measuring flask, add 70% ethanol to the mark, and get it.
Detection method:
Precisely draw 10 µl each of the reference solution and the test solution, and inject it into the HPLC for determination.
This product is calculated as a dry product and contains at least 0.050% luteolin (C21H20O11).