Method for establishing HPLC (high performance liquid chromatography) fingerprint graphic for ointment for unblocking collaterals and relieving pain as well as standard fingerprint graphic thereof
A technology of fingerprints and determination methods, which is applied in the field of determination of fingerprints of traditional Chinese medicines, and achieves the effects of good reproducibility, good specificity, and good coincidence
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Embodiment 1
[0059] 1. Preparation of the test solution: Take an appropriate amount of sample, use 100% methanol solution, ethanol, and 75% ethanol solution as the extraction solvent, ultrasonically extract for 0.5h, and filter the solution with a 0.45um filter membrane. Take the continued filtrate as the test solution for subsequent use.
[0060] 2. HPLC analysis conditions
[0061] Kromasil 100-5C 18 Analytical column (4.6mm×250mm, 5um); mobile phase: A is 0.2% phosphoric acid aqueous solution, B phase is methanol, flow rate is 1.0ml / min; detection wavelength is 220-300nm; column temperature is 30°C, injection 15ul.
[0062] The gradient elution program is as follows:
[0063]
[0064] 3. Determination: Precisely draw the test solution and inject it into the liquid chromatograph, measure according to high performance liquid chromatography, obtain the corresponding atlas, and obtain the HPLC atlas under the corresponding chromatographic conditions.
Embodiment 2
[0066] 1. Preparation of the test solution: Take an appropriate amount of sample, use 75% methanol solution, methanol, ethanol, and 75% ethanol solution as extraction solvents, reflux extraction for 1 hour, and filter the solution with a 0.45um filter membrane. Take the continued filtrate as the test solution for subsequent use.
[0067] 2. HPLC analysis conditions
[0068] Kromasil 100-5C 18 Analytical column (4.6mm×250mm, 5um); mobile phase: A is water, B phase is acetonitrile, flow rate is 1.0ml / min; detection wavelength is 220-300nm; column temperature is 30°C, injection 20ul.
[0069] The gradient elution program is as follows:
[0070]
[0071] 3. Determination: Precisely draw the test solution and inject it into the liquid chromatograph, measure according to high performance liquid chromatography, obtain the corresponding atlas, and obtain the HPLC atlas under the corresponding chromatographic conditions.
Embodiment 3
[0073] 1. Preparation of the test solution: Take an appropriate amount of sample, use 75% methanol solution, methanol, ethanol, and 75% ethanol solution as extraction solvents, reflux extraction for 2 hours, and filter the solution with a 0.45um filter membrane. Take the continued filtrate as the test solution for subsequent use.
[0074] 2. HPLC analysis conditions
[0075] Shiseido MG C 18 (4.6mm×250mm, 5um); mobile phase: phase A is a mixed aqueous solution of 0.2% phosphoric acid and 0.4% triethylamine (v / v), phase B is acetonitrile, and the flow rate is 1.0ml / min; the detection wavelength is 220-300nm; Temperature 30 ℃, inject 20ul.
[0076] The gradient elution program is as follows:
[0077]
[0078] 3. Determination: Precisely draw the solution of the test product and inject it into the liquid chromatograph, measure according to the high performance liquid chromatography, and obtain the corresponding atlas.
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