Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
41, Number 1, January 2003, pp. 36-40
Study on the Determination of Polyphenols in Tobacco by HPLC
Coupled with ESI–MS After Solid-Phase Extraction
Zhong Li[1],[2], Lan Wang1, Guangyu Yang[1], Honglin Shi[1],
Ciqing Jiang[1], Wei Liu[2], and Yunhuai Zhang[2]
[1]Key Laboratory of Chemistry & Engineering, Yunnan Academy of Tobacco
Science, Kunming 650106, P.R. China and
[2]College of Chemistry & Engineering,
Chongqing University, Chongqing 400044, P.R. China
A high-performance liquid chromatography method coupled with
electrospray ionization–mass spectrometry for the determination of polyphenols
in tobacco is studied. The polyphenols are extracted from a tobacco sample
by being refluxed in a boiling water bath with 80% methanol and purified by
solid-phase extraction with a C18 cartridge. The chlorogenic acid, rutin, scopoletin,
caffeic acid, scopolin, and other polyphenols are satisfactorily separated
on a Nova-Pak C18 chromatographic column (3.9 ¥ 150 mm) with methanol and
0.05 mol/L potassium dihydrogen phosphate buffer solution gradient elution
as mobile phase at a flow rate of 0.5 mL/min. Each of the polyphenols is monitored
by photodiode array detector at its maximum wavelength: chlorogenic acid, 326.1
nm; rutin, 354.8 nm; scopoletin, 344.0 nm; caffeic acid, 323.7 nm; and scopolin,
365.2 nm. The limits of detection are: 100 ng/mL for chlorogenic acid, 125
ng/mL for rutin, 60 ng/mL for scopoletin, 50 ng/mL for caffeic acid, and 100
ng/mL for scopolin. The key polyphenols in tobacco are identified by comparing
the retention time, the UV-spectrum, and the mass spectra with those of the
standards. The recovery of tobacco polyphenols is 94–105%, and the relative
standard deviations are 1.28–1.49%. This method is successfully applied
to qualitatively and quantitatively analyze the polyphenols in tobacco with
good results.
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