Method for Establishing HPLC Fingerprint of Yixuan Granules
A fingerprint and method establishment technology, which is applied in the field of drug detection, can solve the problems that the research on fingerprint establishment methods is still relatively small, and achieve the effects of short chromatographic analysis time, good reproducibility and stability, and quick and easy processing
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Embodiment 1
[0066] The optimization of embodiment 1 chromatographic conditions
[0067] The preparation method of the test solution: Weigh 5g of Yixuean granules, add 50mL of water to dissolve, add 0.3mL of dilute hydrochloric acid, stir well, let stand for 20min, filter, add water-saturated n-butanol to extract 5 times, each time 15mL, divide Take n-butanol extracts, combine them, let stand for 1h, separate to obtain n-butanol liquid, recover n-butanol, add 5 mL of methanol to the residue, centrifuge, and take supernatant to obtain the test solution.
[0068] 1-1 Selection of chromatographic column
[0069] The other determination conditions of high performance liquid chromatography are the same. Welchrom C18 column (4.6mm×250mm, 5μm), Hypersil C18 column (4.6mm×250mm, 5μm) and Phenomenex Gemini C18 column (4.6mm×250mm, 5μm) three column, the resulting chromatograms are divided into figure 1 , figure 2 , image 3 shown by image 3 It can be seen that the Phenomenex Gemini C18 colum...
Embodiment 2
[0078] The optimization of embodiment 2 sample pretreatment method
[0079] Chromatographic conditions of high-performance liquid chromatography: the chromatographic column adopts Phenomenex Gemini C18 column (4.6mm × 250mm, 5 μm); the guard column adopts Phenomenex C18 (4 × 3.0mm); mobile phase: methanol-0.1% glacial acetic acid; gradient elution Procedure: Time Gradient
[0080] 0min→11min→16min→29min→30min→35min→45min→55min→70min→95min; the corresponding gradient elution solution is composed of mobile phase A methanol and mobile phase B 0.1% glacial acetic acid according to the following volume ratio, mobile phase A Volume percentage gradient 5%→16%→17%→30%→34%→34%→35%→45%→45%→70%, mobile phase B volume percentage gradient 95%→84%→83%→70 %→66%→66%→65%→55%→55%→30%;; detection wavelength: 256nm; flow rate: 1.0mL·min-1; column temperature: 25°C; injection volume: 5μL.
[0081] 2-1 Selection of extraction solvent
[0082] Weigh 5g of Yixuan granules, select water, methanol a...
Embodiment 3
[0089] Example 3 Establishment of HPLC Fingerprint of Yixuan Granules
[0090] 3-1 Preparation of reference substance solution: Weigh 0.81mg of protocatechuic acid standard substance and place it in a 10mL volumetric flask, add an appropriate amount of methanol to dissolve it and set the volume to the mark, shake well, and obtain a solution with a concentration of 0.081mg·mL-1 Protocatechuic acid standard solution
[0091]3-2 Preparation of the test solution: Weigh 5g of Yixuan granules, add 50mL of water to dissolve, add 0.3mL of dilute hydrochloric acid, stir well, let stand for 20min, filter, add water and extract with n-butanol for 5 times, 15mL each time , separate the n-butanol extracts, combine them, let stand for 1h, separate and obtain the n-butanol liquid, recover the n-butanol, add methanol 5mL to the residue, centrifuge, and take the supernatant to obtain the test solution.
[0092] 3-3 Chromatographic conditions of high performance liquid chromatography:
[0093...
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