Determination method of effective constituents of pharmaceutical composition for invigorating spleen and strengthening mid-energy as well as dispelling cold and checking diarrhea, and method for establishing fingerprint spectrum
A determination method and a technique for strengthening the spleen and warming the medium are applied in the field of high-performance liquid chromatography detection of Chinese patent medicines, which can solve the problems of large consumption of organic solvents, non-uniformity, lack of overall quality control of medicines, etc., and achieve good and accurate separation of main peaks. high degree of effect
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Embodiment 1
[0060] Embodiment 1 Fingerprint Atlas Establishment
[0061] 1.1 Instrument
[0062]Agilent 1260 HPLC, Shimadzu LC-20AT HPLC, Agilent1200 HPLC, column (Agilent ZORBAX SB-C18, 4.6×250mm, 5μm, S / N: USCL 060354), chromatography Column (Agilent ZORBAX SB-C18, 4.6×250mm, 5μm, S / N:USCL 058060), chromatography column (Agilent ZORBAX SB-C18, 4.6×250mm, 5μm, S / N:USCL 036549), chromatography column (Agilent ZORBAX SB -C18, 4.6×250mm, 5μm, S / N:USCL 059648), chromatographic column (Phenomenex Gemiri C18, 4.6×250mm, 5μm, S / N:695685-14), chromatographic column (Thermo ODS-2HYPERSIL, 4.6×250mm , 5μm, S / N:0905415N4), chromatographic column (Inertsil ODS-3, 4.6×250mm, 5μm, S / N:1A7143425), chromatographic column (Unitary C18, 4.6×250mm, 5μm, S / N:Z2013112901).
[0063] 1.2 Reagents
[0064] Reagents: formic acid (AR, Sinopharm Chemical Reagent Co., Ltd.), glacial acetic acid (AR, Sinopharm Chemical Reagent Co., Ltd.), phosphoric acid (AR, Beijing Jingqiu Chemical Co., Ltd.), pure water (Hangz...
Embodiment 2
[0077] Embodiment 2 Optimal chromatographic conditions investigation
[0078] Apparatus and reagents are the same as in Example 1.
[0079] 2.1 Selection of detection wavelength
[0080] A PDA detector was used to detect the test product, investigate the chromatographic peak information of the sample, and screen the detection wavelength.
[0081] Preparation of the test solution: take this product, cut into pieces, and mix well. Take 1g, accurately weigh it, put it in a stoppered Erlenmeyer flask, accurately add 50ml of 50% methanol, seal it tightly, weigh it, ultrasonically treat it (500W, 40KHZ) for 30 minutes, take it out, put it at room temperature, and make up with 50% methanol Subtract the lost weight, shake well, filter, take the continued filtrate, that is.
[0082] Agilent ZORBAX SB-C18 chromatographic column was used, the column temperature was 30°C, the flow rate was 0.8ml / min, methanol was used as mobile phase A, and aqueous solution was used as mobile phase B, ...
Embodiment 3
[0122] Embodiment 3 comparative study of mobile phase system
[0123] According to the pre-experimental results, different mobile phase systems were investigated, and the chromatographic conditions with the most abundant chromatographic peak information and the best resolution were screened.
[0124] Preparation of the test solution: take this product, cut into pieces, and mix well. Take 1g, accurately weigh it, put it in a stoppered Erlenmeyer flask, accurately add 50ml of 50% methanol, seal it tightly, weigh it, ultrasonically treat it (500W, 40KHZ) for 30 minutes, take it out, put it at room temperature, and make up with 50% methanol Subtract the lost weight, shake well, filter, take the continued filtrate, that is.
[0125] Using Agilent ZORBAX SB-C18 chromatographic column, column temperature 30 ℃, detection wavelength 254nm, flow rate 1.0ml / min, investigate acetonitrile-water solution, methanol-water solution, methanol-0.2% formic acid solution, methanol-0.2% phosphoric...
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