| This paper studies 200 samples supplied by ten cooperators in order to establish fingerprint chromatography of tobacco liquid system.After comparing liquid-liquid extraction,freeze-dry-dissolve and dilution, dilution is adopted as pre-treatment method finally.Inner-standard(IS) is used to determine relative quality during the method-searching stage. Relative retention time(RRT) RSD and Relative peak aera(RPA) RSD for accuracy experiment are respectively under 0.19% and 6.60%;RRT RSD and RPA RSD for reiteration experiment are respectively under 0.58% and 4.51%; RRT RSD and RPA RSD for stability experiment are respectively under 6.89% and 9.6%.As a discussion result, inter-standard is changed into outer-standard method. 2-methylnaphthalene is used as outer-standard, work curve is y=24512x+127298, R~2=0.9964. Cromatogram column model is Symmetry C18 4.6mm(ID)*250mm 5μm; Column temperature is 40℃; A-Acetonitrile, B-0.1%Formic Acid are used as liquid phase; Determine wavelength is 254nm; Flow is 1.0mL/min; Gradient condition: 0～80min: 100%B, 80～99min: 100A, 99～100min: 100%B. Under said condition, samples about different directions,storages and batches are studied, and great amount of data is obtained.In study of aromatic compounds, nitrobenzene is chosen as inner-standard. HPLC condition is the same as outer-standard method. Quality and reclaiming rate of vanillin,3-ethoxy-4-hydroxybenzaldehyde,trans-cinnamic acid,benzyl cinnamate and p-anisaldehyde in the sample are determined. Said standard matters work curves respectively are: y=13098x+193680, R~2=0.9999; y=12544x+83758, R~2=0.9999; y=49868x+717351, R~2=0.9985; y=34254x+330037, R~2=0.9998; y=28020x+255897, R~2=0.9997.Said five matters' lowest determine limits respectively are: 13.8,16.5,3.4,0.6,8.1ug/g. RSD for reiteration experiment is 0.21%-1.31%. Reclaiming rate of said standard matters is 100.6%-11.8%... |
PDF Full Text Request