HPLC finger-print detection method of Heracleum souliei
A Kangding independent living and detection method technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as differences in drug efficacy, and achieve high precision, good separation, and good repeatability
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Embodiment 1
[0048] Embodiment 1 The HPLC fingerprint spectrum detection method of Kangding Duhuo of the present invention
[0049] (1) Preparation of the reference solution: Accurately weigh 0.5g of Kangding Duhuo powder, place it in a 50mL conical flask, add 30mL of methanol, weigh after cold soaking for 60min, ultrasonic (power 200W, frequency 40KHZ) for 30min, take out, Let cool, add weight, and take the supernatant to pass through a 0.45 μm microporous membrane as a reference solution;
[0050] (2) Preparation of reference solution: accurately weigh appropriate amounts of psoralen reference substance and bergamot lactone reference substance, mix them, and use a 10mL volumetric flask to constant volume to prepare psoralen and bergamot lactone concentrations of 0.086mg / mL and 0.358mg / mL methanol solutions, as the reference solution;
[0051] (3) Establishment of contrast fingerprints:
[0052] The reference substance solution and the reference substance solution prepared from 1-10 bat...
Embodiment 2
[0059] Example 2 Conditional screening and methodological investigation of the detection method of the present invention
[0060] The test article under the present embodiment is selected from the medicinal materials of Kangding Independence in Table 1.
[0061] 1 Chromatographic conditions
[0062] Using DiamonilC 18 Chromatographic column (250×4.6mm, 5μm), the mobile phase A is water, the mobile phase B is methanol, the column temperature is 30°C, the flow rate is 1mL / min, the detection wavelength is 309nm, the inspection time is 50min, and the injection volume is 20μL. Gradient elution: 45% B for 5 minutes, 45%-60% B for 5 minutes, 60%-73% B for 25 minutes, 73%-80% B for 5 minutes, and 80% B for 10 minutes, namely:
[0063]
[0064] 1.1 Selection of detection wavelength
[0065] Adopt diode array detector, carry out full-wavelength scanning to need testing solution, compare HPLC-3D figure under each wavelength, the result is as follows Figure 4 shown.
[0066] The ...
Embodiment 3
[0124] Embodiment 3 The present invention measures the method for psoralen and bergamot lactone in Kangding Duhuo
[0125] 1 Linear relationship investigation
[0126] Accurately weigh the appropriate amount of psoralen reference substance and bergamot lactone reference substance, mix them, and use a 10mL volumetric flask to make up the concentration of psoralen and bergamot lactone to be 0.086mg / mL and 0.358mg / mL respectively methanol solution to obtain a mixed reference solution of bergamotide and psoralen.
[0127] Precisely draw 0.4, 0.8, 1, 4, 8, 12, 20 μL of the mixed reference solution of bergamot lactone and psoralen, and measure according to the chromatographic conditions in Example 1, with the injection volume as the abscissa and the peak area as the ordinate Coordinates, the regression equations are Y=3540.5X+339.93 (R 2 =0.9982), Y=2421.0X-23.790 (R 2 =0.9982), the linear ranges were 0.1432-7.16mg and 0.0344-1.72mg, respectively. The result is as Figure 10 an...
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