Automated analysis of the serum antioxidative activities against five different reactive oxygen species by sequential injection system with a chemiluminescence detector.
BACKGROUND
There is a growing evidence that reactive oxygen species (ROS) may cause many pathologic conditions including chronic diseases, neurodegenerative disorders, cancer and aging. There are a number of methods to measure the total antioxidative activity of the serum. However, since the lifetime and oxidative activity of various types of ROS are all different, to measure simply the total antioxidative activity of the serum is not enough. Therefore, to aid the diagnosis and to improve the therapeutic strategy, it is important to develop a simple and reliable method of assaying antioxidative activity of the serum.


METHODS
A method that combines sequential injection analysis (SIA) and luminol chemiluminescence (CL) detection was employed for the measurement of antioxidative activities of human serum. We collected sera from healthy subjects (n=42) and patients with diabetes (n=39) and rheumatoid arthritis (n=25) and tested the sensitivity, reproducibility and reliability of our method.


RESULTS
Since the operation is automatically controlled by a personal computer, we obtained a satisfactory repeatability without the need of much manpower. The time required for obtaining the antioxidative activity against one ROS for one individual is less than 3min. Although the value of antioxidative activity varies from subject to subject, there were a certain relationship between the disease and the antioxidative values of each type of ROS. The results suggest that the measurement of antioxidative activity against different ROS may provide us with valuable information regarding the disease state.


CONCLUSIONS
The evaluation of antioxidative activities against each ROS by the proposed method should be more informative to understand the antioxidative status of biological fluid.
Kishikawa
Naoya
N
Department of Environmental and Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
Ohyama
Kaname
K
Yao
Junko
J
Miyamoto
Aoi
A
Imazato
Takahiro
T
Ueki
Yukitaka
Y
Nakashima
Kenichiro
K
Maehata
Eisuke
E
Kuroda
Naotaka
N
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000975:Antioxidants / Q000032:analysis* / Q000378:metabolism*; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000378:metabolism; D001331:Automation; D001774:Blood Chemical Analysis / Q000379:methods*; D003920:Diabetes Mellitus / Q000097:blood / Q000378:metabolism; D005260:Female; D006801:Humans; D007267:Injections; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D008875:Middle Aged; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015203:Reproducibility of Results; D055815:Young Adult
D000975:Antioxidants; D017382:Reactive Oxygen Species