A method to identify whether pinellia is mixed with tiger palm southern star
A technology of Southern Tiger Palm and Pinellia, which is applied in the field of identifying whether Pinellia is mixed with Southern Star Tiger Palm, so as to improve the quality control level and guide the effect
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[0027] Preparation of relevant solutions in the test
[0028] Denaturation buffer: Weigh 573.1g of guanidine hydrochloride, 121.1g of tris, 0.734g of ethylenediaminetetraacetic acid, add water to dissolve, add concentrated hydrochloric acid to adjust the pH to 8.0, add water to dilute to 1L, and shake well.
[0029] DTT solution: Weigh an appropriate amount of DTT (dithiothreitol), dissolve it in water, and get it (concentration is 0.50M);
[0030] IAA solution: Weigh an appropriate amount of IAA (iodoacetamide), dissolve it in water, and get it (concentration is 0.55M, ready to use);
[0031] Ammonium bicarbonate solution: Weigh an appropriate amount of ammonium bicarbonate and dissolve it in water to obtain (concentration: 1.0%);
[0032] Acetic acid solution: Accurately measure an appropriate amount of acetic acid and dilute it with water to get it (concentration is 0.5%);
[0033] Bovine trypsin solution: Weigh an appropriate amount of bovine trypsin, dissolve it in acet...
Embodiment 1
[0035] 1 Instruments, reagents and samples
[0036] Instruments: Waters Quattro Premier XE high performance liquid chromatography-mass spectrometry; SartoriusCP225D electronic balance.
[0037] Reagents: trypsin (manufactured by Sigma, batch number: SLBG6452V); dithiothreitol (DTT), iodoacetamide (IAA), ammonium bicarbonate, and acetic acid were all of analytical grade.
[0038] Sample: pinellia herbal medicine (collected in the market). Pinellia control medicinal materials (provided by the China Inspection Institute, batch number: 121272-201404 and 121272-201605).
[0039] Liquid phase and mass spectrometry conditions
[0040] Liquid phase conditions: the column is ACQUITY UPLC ® BEH C 18(2.1×100mm, 1.7μm), column temperature 35°C, flow rate 0.3mL / min, mobile phase A is 0.1% formic acid solution, B is 0.1% formic acid acetonitrile solution, gradient elution is performed, see the table below.
[0041] Table 1 Gradient elution program
[0042]
[0043] Mass spectromet...
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