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. 2012 Jan;402(3):1389-92.
doi: 10.1007/s00216-011-5563-5. Epub 2011 Nov 26.

Interference of flavonoids with enzymatic assays for the determination of free fatty acid and triglyceride levels

Elise F Hoek-van den Hil  1 , Karsten Beekmann, Jaap Keijer, Peter C H Hollman, Ivonne M C M Rietjens, Evert M van Schothorst
Affiliations

Interference of flavonoids with enzymatic assays for the determination of free fatty acid and triglyceride levels

Elise F Hoek-van den Hil et al. Anal Bioanal Chem. 2012 Jan.

Abstract

Flavonoids are bioactive food compounds with potential lipid-lowering effects. Commercially available enzymatic assays are widely used to determine free fatty acid (FFA) and triglyceride (TG) levels both in vivo in plasma or serum and in vitro in cell culture medium or cell lysate. However, we have observed that various flavonoids interfere with peroxidases used in these enzymatic assays, resulting in incorrect lower FFA and TG levels than actually present. Furthermore, addition of isorhamnetin or the major metabolite of the flavonoid quercetin in human and rat plasma, quercetin-3-O-glucuronide, to murine serum also resulted in a significant reduction of the detected TG levels, while a trend was seen for FFA levels. It is concluded that when applying these assays, vigilance is needed and alternative analytical methods, directly assessing FFA or TG levels, should be used for studying the biological effects of flavonoids on FFA and TG levels.

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Figures

Figure
Figure
Interference of different flavonoids spiked into cell culture medium on the FFA (A) and TG (B) levels, and interference of quercetin (Q) and its major metabolite, quercetin-3-O-glucuronide (Q3G), and isorhamnetin (I) spiked into murine serum on the FFA (C) and TG (D) levels, as measured by enzymatic assays. Flavonoids analysed are quercetin (Q), (+)-catechin (C), luteolin (L), genistein (G), naringenin (N), and kaempferol (K). Asterisk indicates a significant difference (p < 0.05) from the control (0 μM)
Fig. 1
Fig. 1
Interference of different flavonoids on the FFA (a) and TG (b) levels in the cell culture medium measured by enzymatic assays. (+)-Catechin (filled square), quercetin (empty square), kaempferol (filled triangle), luteolin (empty triangle), genistein (filled circle), and naringenin (empty circle). Results are mean ± SEM, n = 3 for FFA and n = 4 for TG. Asterisk indicates a significant difference (p < 0.05) from the control (0 μM). b Quercetin has already a significant effect at 10 μM
Fig. 2
Fig. 2
Interference of quercetin (empty square) and its major metabolite, quercetin-3-O-glucuronide (filled square), and isorhamnetin (circle) in murine serum on the FFA (a) and TG (b) levels measured by enzymatic assays. Results are mean ± SEM, n = 8. Asterisk indicates a significant difference (p < 0.05) from the control (0 μM)
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