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Am. J. Biomed. Sci. 2013, 5(2), 154-160; doi: 10.5099/aj130200154
Received: 5 February 2013; | Revised: 26 February 2013; | Accepted: 28 March 2013


A Novel Approach for Measurement of Total Reactive Oxidant Species (ROS) In Vivo by A Fluorometric Method


Boris Nemzer1, Zbigniew Pietrzkowski2, Tony Chang3, and Boxin Ou4

1FutureCeuticals, 300 West Sixth Street, Momence, IL 60954, USA

2Applied BioClinical Inc., 16259 Laguna Canyon Rd, Irvine, CA, 92618, USA

3International Chemistry Testing, 258 Main Street, Suite 311, Milford, MA 01757, USA

4Dover Sciences, 7 Dover Circle, Franklin, MA 02038, USA

*Corresponding author:

Boxin Ou

Dover Sciences

7 Dover Circle

Franklin, MA 02038


Email: boxinou@gmail.com



Six healthy participants were given a single dose of 100 mg of CoffeeBerry® whole coffee fruit extract containing high levels of antioxidants to verify an acute effect of the treatment on ROS serum level. Blood samples were collected at 0 min, 60 min, 120 min and 180 min for subsequent measurements of serum ROS level using dihydrorhodamine 6G (DHR6G) as a fluorescent probe. The nonfluorescent DHR6G, after being oxidized by ROS present in serum samples, became rhodamine 6G (R6G) and emitted fluorescence. By quantifying R6G specific fluorescence, we were able to measure the ROS concentration. DHR 6G is indiscriminate to various free radicals (FR) found in the human body, thus DHR 6G can be very useful in quantifying total ROS in vivo. Our data indicated that five participants responded to the intake of CoffeeBerry® whole coffee fruit extract by significant decrease of ROS concentrations in vivo. Collected results are promising and indicating that DHR6G-based method could be reliable and efficacious to measure acute serum ROS changes in response to single dose treatment with antioxidant products. Therefore further clinical validation of this test is justified.

Keywords: Free radicals, CoffeeBerry®, Fluorescence Probe, DHR6G, Rhodamine 6G.

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