Tandem mass spectrometry studies of green tea catechins. Identification of three minor components in the polyphenolic extract of green tea.
Liquid chromatography/electrospray ionization mass and tandem mass spectrometry (MS/MS) techniques were used to identify two minor components and one new compound in the polyphenolic extract of green tea (Camellia sinensis). Identification and structure assignments were based on previously reported sub-structural features in the MS/MS product, precursor and neutral loss scans of reference samples. The structures of two minor components, related to the known green tea components epicatechin gallate (ECG, 5) and epigallocatechin gallate (EGCG, 6), are formed by methylation at the 3"-O-position of the gallic acid moiety. The new compound contained a gallic acid ester group, but had only one phenolic group in either the A- or B-ring, relative to the structure of 5. High-resolution mass measurements supported the empirical formula assigned to the new compounds. An important fragmentation for defining the position of methylation of the ester function involves ionization of the phenolic group at the 4"-position of the gallic acid, followed by elimination of the ester function as a neutral with concomitant formation of the m/z 169 ion. If the 4"-position is blocked by methylation, the formation of m/z 169 incorporating the gallic acid group would be blocked. Thus, the presence of an ion representing the ester group indicates a free 4"-phenol and the absence of this ion would signify the 4"-position as a site of methylation. The operation of this mechanism should be general and useful in assigning the site of methylation of any polyphenolic ester group in natural products. A similar conclusion can be drawn concerning alkylation or esterification of the 4'-position of the catechins, i. e. blocking the 4'-phenol would prevent formation of the m/z 125 ion common to all of the catechin compounds thus far examined. Therefore, mass spectral data are of considerable importance in deducing the sites of alkylation or esterification in the structures of the components of green tea.
Division of Medicinal Chemistry, College of Pharmacy, University of Arizona, Tucson 85721, USA.
P
P
Miketova
K H
KH
Schram
J
J
Whitney
M
M
Li
R
R
Huang
E
E
Kerns
S
S
Valcic
B N
BN
Timmermann
R
R
Rourick
S
S
Klohr
D002392:Catechin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000302:isolation & purification; D005419:Flavonoids; D013058:Mass Spectrometry / Q000379:methods; D008968:Molecular Conformation; D010636:Phenols / Q000737:chemistry* / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry; D011108:Polymers / Q000737:chemistry* / Q000302:isolation & purification; D013662:Tea / Q000737:chemistry*
D005419:Flavonoids; D010636:Phenols; D010936:Plant Extracts; D011108:Polymers; D013662:Tea; D002392:Catechin