Method for authenticating dendrobium officinale by using HPLC fingerprint
A technology of Dendrobium officinale and fingerprint spectrum, which is applied in the field of establishment of fingerprints of Chinese medicinal materials, can solve the problems of staying, unfavorable to standardize the management of the traditional Chinese medicine industry, unable to realize the quality control of the medicinal materials of Dendrobium officinale, etc., and achieves the effects of easy operation and good reproducibility.
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Embodiment 1
[0037] 1. Instruments and reagents
[0038] 1.1 Instrument: Agilent 1200 high performance liquid chromatograph; DAD detector.
[0039] 1.2 Reagents: Dendrobium officinale and its counterfeit products or substitutes are commercially available or provided by manufacturers; acetonitrile and methanol are chromatographically pure, and the rest of the reagents are analytically pure.
[0040] 2. Establishment of standard fingerprint of Dendrobium officinale medicinal materials
[0041] (1) Preparation of the test solution: Precisely weigh 10.0 g of fresh Dendrobium officinale medicinal materials, cut them into pieces, add 200 mL of water to reflux for extraction for 1 h, recover to a certain volume under reduced pressure, and extract with water-saturated n-butanol (20 mL*3 times), Recover n-butanol, dilute to volume with methanol, filter with 0.22 μm membrane, and use the filtrate as the test solution.
[0042] (2) Selection of chromatographic conditions: chromatographic separation...
Embodiment 2
[0047] The difference from Example 1 is:
[0048] Establishment of standard fingerprint of Dendrobium officinale:
[0049] (1) Preparation of the test solution: accurately weigh 2.0g of Dendrobium candidum, cut it into pieces, add 100mL of water to reflux for extraction for 1.5h, recover to a certain volume under reduced pressure, extract with ether (30mL*2 times), and recover ether , Methanol constant volume, 0.22μm membrane filtration, filtrate as the test solution.
[0050] (2) Selection of chromatographic conditions: the chromatographic separation column is SB-C18; the mobile phase is methanol-0.2% phosphoric acid; the gradient elution method is 0min→5min→12min→25min→40min→60min→70min; methanol 7%→14% →22%→30%→45%→55%→78%; flow rate 1.0mL / min; detection wavelength 270nm; column temperature 25°C; analyze the test solution under these conditions to obtain the fingerprint of Dendrobium candidum medicinal material.
Embodiment 3
[0052] The difference from Example 1 is:
[0053] Establishment of Standard Fingerprint of Dendrobium officinale
[0054] (1) Preparation of the test solution: Accurately weigh 1.0 g of Dendrobium candidum medicinal material powder, add 50 mL of methanol for ultrasonic extraction for 30 min, recover to a certain volume under reduced pressure, extract with ethyl acetate (20 mL*3 times), and recover ethyl acetate , Methanol constant volume, 0.45μm membrane filtration, filtrate as the test solution.
[0055] (2) Selection of chromatographic conditions: the chromatographic separation column is Xtimate TM C18; mobile phase is methanol-0.2% phosphoric acid; gradient elution method 0min→5min→10min→25min→45min→60min→75min; methanol 25%→25%→30%→40%→55%→70%→100 %; Flow rate 1.0mL / min; Detection wavelength 280nm; Column temperature 30°C; Analyze the test solution under this condition to obtain the fingerprint of Dendrobium candidum medicinal material.
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