High-performance liquid chromatography with coulometric electrode array detector for the determination of quercetin levels in cells of the immune system

被引:50
作者
Bugianesi, R
Serafini, M
Simone, F
Wu, DY
Meydani, S
Ferro-Luzzi, A
Azzini, E
Maiani, G [1 ]
机构
[1] Natl Inst Food & Nutr Res, Human Nutr Unit, Antioxidant Res Lab, I-00178 Rome, Italy
[2] Tufts Univ, Nutr Immunol Lab, Jean Mayer Human Nutr Res Ctr Aging, Boston, MA 02111 USA
关键词
quercetin; electrochemical detection; liquid chromatography; splenocytes; antioxidant; flavonoids; cells;
D O I
10.1006/abio.2000.4697
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
There is mounting evidence emphasizing the importance of intracellular antioxidant levels for maintenance of the immune function. The flavonoid quercetin, a natural antioxidant, has been shown to modulate enzymes involved in the regulation of the inflammatory response. However, up to now, there have been no studies describing quercetin levels in cells of the immune system. A gradient reversed-phase HPLC technique to identify and quantify intracellular levels of quercetin and its application in mice splenocytes are described. Mobile phases were a 0.01 M sodium phosphate monobasic solution adjusted to pH 2.8 with 85% orthophosphoric acid (buffer, Solvent A) and methanol (Solvent B) with a dow rate of 1 ml/min. An eight-channel coulometric electrode array detector was used. In vitro supplementation with increasing concentration of quercetin (25, 50, and 100 mu M) raises intracellular quercetin levels in a dose-dependent manner. The method has the required features of specificity and sensitivity for monitoring quercetin uptake in cells of the immune system. (C) 2000 Academic Press.
引用
收藏
页码:296 / 300
页数:5
相关论文
共 19 条
[1]   FLAVONOIDS AND RELATED-COMPOUNDS AS INHIBITORS OF ARACHIDONIC-ACID PEROXIDATION [J].
BAUMANN, J ;
BRUCHHAUSEN, FV ;
WURM, G .
PROSTAGLANDINS, 1980, 20 (04) :627-639
[2]   Validation of chromatographic methods in biomedical analysis - Viewpoint and discussion [J].
Causon, R .
JOURNAL OF CHROMATOGRAPHY B, 1997, 689 (01) :175-180
[3]   INHIBITION OF CONSTITUTIVE ENDOTHELIAL NO-SYNTHASE ACTIVITY BY TANNIN AND QUERCETIN [J].
CHIESI, M ;
SCHWALLER, R .
BIOCHEMICAL PHARMACOLOGY, 1995, 49 (04) :495-501
[4]   PROTEIN-KINASE C INHIBITION BY PLANT FLAVONOIDS - KINETIC MECHANISMS AND STRUCTURE-ACTIVITY-RELATIONSHIPS [J].
FERRIOLA, PC ;
CODY, V ;
MIDDLETON, E .
BIOCHEMICAL PHARMACOLOGY, 1989, 38 (10) :1617-1624
[5]   ANALYSIS OF PHENOLIC AND FLAVONOID COMPOUNDS IN JUICE BEVERAGES USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH COULOMETRIC ARRAY DETECTION [J].
GAMACHE, P ;
RYAN, E ;
ACWORTH, IN .
JOURNAL OF CHROMATOGRAPHY, 1993, 635 (01) :143-150
[6]  
GAMACHE PH, 1993, CLIN CHEM, V39, P1825
[7]  
GERRITSEN ME, 1995, AM J PATHOL, V147, P278
[8]   AN ANALYSIS OF THE WASHINGTON CONFERENCE REPORT ON BIOANALYTICAL METHOD VALIDATION [J].
HARTMANN, C ;
MASSART, DL ;
MCDOWALL, RD .
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, 1994, 12 (11) :1337-1343
[9]   Validation of bioanalytical chromatographic methods [J].
Hartmann, C ;
Smeyers-Verbeke, J ;
Massart, DL ;
McDowall, RD .
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, 1998, 17 (02) :193-218
[10]   Application of a new high-performance liquid chromatographic method for measuring selected polyphenols in human plasma [J].
Maiani, G ;
Serafini, M ;
Salucci, M ;
Azzini, E ;
FerroLuzzi, A .
JOURNAL OF CHROMATOGRAPHY B, 1997, 692 (02) :311-317