Improved superoxide-generating system suitable for the assessment of the superoxide-scavenging ability of aqueous extracts of food constituents using ultraweak chemiluminescence

Chin Hung Tsai, Rey Chang Chang, Jeng Fong Chiou, Tsan Zon Liu

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33 Citations (Scopus)

Abstract

In the interest of developing a simple and rapid ultraweak chemiluminescence assay for assessing the superoxide (O2 -)-scavenging activities of various aqueous extracts of food constituents, a specific and stable O2 --generating system was sought. Reported herein is the obtainment for the first time of a specific and stable O2 --generating system consisting of methylglyoxal (MG), a reactive 2-oxo aldehyde and arginine, which has been shown to produce much steadier lucigenin-based chemiluminesence (LBCL) than the conventional xanthine/xanthine oxidase system running in parallel and monitoring by an ultraweak chemiluminescence analyzer. Upon mixing of MG and arginine in a phosphate-buffered saline solution, pH 7.4, steady, time-dependent increments of LBCL can be visually observed. The plateau of LBCL can be reached in approximately 10 min and retained in a steadily stable state thereafter without fluctuation for the next 15 min. The lucigenin-based LBCL generation was shown to be specific since it could be effectively inhibited by active bovine SOD, but not by heat-inactivated enzyme or catalase. Conversely, the xanthine/xanthine oxidase system can merely produce a LBCL peak rapidly but decay instantaneously. To illustrate the application of the proposed method for assessing the O2 --scavenging ability of various food extracts, namely, Prunus mume (A), Lilum lancifolium (B), Creataegus pinnatifida (C), Tremella fuciformis (D), Fortunella margarita (E), and Scutellaria baicalensis (F), we used the following protocol: 12 min after monitoring of LBCL, 1 mg/mL of each of the test compounds was added to the assay system and various degrees of sudden drop of LBCL values were observed, indicating differences in O2 --scavenging abilities exerted by these food extracts that can be visually compared. Consequently, the percentages of inhibition of LBCL versus the concentrations of a test compound can be constructed. It follows that the concentration needed to inhibit 50% of LBCL (IC50) of a test compound can be extrapolated from the curve. Using this approach, we were able to obtain the IC50 values of various compounds to be tested and the order of inhibitory efficiency of the above-mentioned food extracts was ranked, being A > B > C > D > E > F, respectively.

Original languageEnglish
Pages (from-to)58-62
Number of pages5
JournalJournal of Agricultural and Food Chemistry
Volume51
Issue number1
DOIs
Publication statusPublished - Jan 1 2003

Fingerprint

Chemiluminescence
chemiluminescence
Scavenging
Luminescence
Superoxides
superoxide anion
Food
xanthine
xanthine oxidase
extracts
arginine
inhibitory concentration 50
Fortunella margarita
Tremella fuciformis
water
Scutellaria baicalensis
Prunus mume
monitoring
testing
Pyruvaldehyde

Keywords

  • Arginine
  • Chemiluminescence emitter
  • Food constituents
  • Methylglyoxal
  • Superoxide-scavenging ability

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)

Cite this

@article{606ccd75d3b74c3ba9c24617612df104,
title = "Improved superoxide-generating system suitable for the assessment of the superoxide-scavenging ability of aqueous extracts of food constituents using ultraweak chemiluminescence",
abstract = "In the interest of developing a simple and rapid ultraweak chemiluminescence assay for assessing the superoxide (O2 -)-scavenging activities of various aqueous extracts of food constituents, a specific and stable O2 --generating system was sought. Reported herein is the obtainment for the first time of a specific and stable O2 --generating system consisting of methylglyoxal (MG), a reactive 2-oxo aldehyde and arginine, which has been shown to produce much steadier lucigenin-based chemiluminesence (LBCL) than the conventional xanthine/xanthine oxidase system running in parallel and monitoring by an ultraweak chemiluminescence analyzer. Upon mixing of MG and arginine in a phosphate-buffered saline solution, pH 7.4, steady, time-dependent increments of LBCL can be visually observed. The plateau of LBCL can be reached in approximately 10 min and retained in a steadily stable state thereafter without fluctuation for the next 15 min. The lucigenin-based LBCL generation was shown to be specific since it could be effectively inhibited by active bovine SOD, but not by heat-inactivated enzyme or catalase. Conversely, the xanthine/xanthine oxidase system can merely produce a LBCL peak rapidly but decay instantaneously. To illustrate the application of the proposed method for assessing the O2 --scavenging ability of various food extracts, namely, Prunus mume (A), Lilum lancifolium (B), Creataegus pinnatifida (C), Tremella fuciformis (D), Fortunella margarita (E), and Scutellaria baicalensis (F), we used the following protocol: 12 min after monitoring of LBCL, 1 mg/mL of each of the test compounds was added to the assay system and various degrees of sudden drop of LBCL values were observed, indicating differences in O2 --scavenging abilities exerted by these food extracts that can be visually compared. Consequently, the percentages of inhibition of LBCL versus the concentrations of a test compound can be constructed. It follows that the concentration needed to inhibit 50{\%} of LBCL (IC50) of a test compound can be extrapolated from the curve. Using this approach, we were able to obtain the IC50 values of various compounds to be tested and the order of inhibitory efficiency of the above-mentioned food extracts was ranked, being A > B > C > D > E > F, respectively.",
keywords = "Arginine, Chemiluminescence emitter, Food constituents, Methylglyoxal, Superoxide-scavenging ability",
author = "Tsai, {Chin Hung} and Chang, {Rey Chang} and Chiou, {Jeng Fong} and Liu, {Tsan Zon}",
year = "2003",
month = "1",
day = "1",
doi = "10.1021/jf020799t",
language = "English",
volume = "51",
pages = "58--62",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society",
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TY - JOUR

T1 - Improved superoxide-generating system suitable for the assessment of the superoxide-scavenging ability of aqueous extracts of food constituents using ultraweak chemiluminescence

AU - Tsai, Chin Hung

AU - Chang, Rey Chang

AU - Chiou, Jeng Fong

AU - Liu, Tsan Zon

PY - 2003/1/1

Y1 - 2003/1/1

N2 - In the interest of developing a simple and rapid ultraweak chemiluminescence assay for assessing the superoxide (O2 -)-scavenging activities of various aqueous extracts of food constituents, a specific and stable O2 --generating system was sought. Reported herein is the obtainment for the first time of a specific and stable O2 --generating system consisting of methylglyoxal (MG), a reactive 2-oxo aldehyde and arginine, which has been shown to produce much steadier lucigenin-based chemiluminesence (LBCL) than the conventional xanthine/xanthine oxidase system running in parallel and monitoring by an ultraweak chemiluminescence analyzer. Upon mixing of MG and arginine in a phosphate-buffered saline solution, pH 7.4, steady, time-dependent increments of LBCL can be visually observed. The plateau of LBCL can be reached in approximately 10 min and retained in a steadily stable state thereafter without fluctuation for the next 15 min. The lucigenin-based LBCL generation was shown to be specific since it could be effectively inhibited by active bovine SOD, but not by heat-inactivated enzyme or catalase. Conversely, the xanthine/xanthine oxidase system can merely produce a LBCL peak rapidly but decay instantaneously. To illustrate the application of the proposed method for assessing the O2 --scavenging ability of various food extracts, namely, Prunus mume (A), Lilum lancifolium (B), Creataegus pinnatifida (C), Tremella fuciformis (D), Fortunella margarita (E), and Scutellaria baicalensis (F), we used the following protocol: 12 min after monitoring of LBCL, 1 mg/mL of each of the test compounds was added to the assay system and various degrees of sudden drop of LBCL values were observed, indicating differences in O2 --scavenging abilities exerted by these food extracts that can be visually compared. Consequently, the percentages of inhibition of LBCL versus the concentrations of a test compound can be constructed. It follows that the concentration needed to inhibit 50% of LBCL (IC50) of a test compound can be extrapolated from the curve. Using this approach, we were able to obtain the IC50 values of various compounds to be tested and the order of inhibitory efficiency of the above-mentioned food extracts was ranked, being A > B > C > D > E > F, respectively.

AB - In the interest of developing a simple and rapid ultraweak chemiluminescence assay for assessing the superoxide (O2 -)-scavenging activities of various aqueous extracts of food constituents, a specific and stable O2 --generating system was sought. Reported herein is the obtainment for the first time of a specific and stable O2 --generating system consisting of methylglyoxal (MG), a reactive 2-oxo aldehyde and arginine, which has been shown to produce much steadier lucigenin-based chemiluminesence (LBCL) than the conventional xanthine/xanthine oxidase system running in parallel and monitoring by an ultraweak chemiluminescence analyzer. Upon mixing of MG and arginine in a phosphate-buffered saline solution, pH 7.4, steady, time-dependent increments of LBCL can be visually observed. The plateau of LBCL can be reached in approximately 10 min and retained in a steadily stable state thereafter without fluctuation for the next 15 min. The lucigenin-based LBCL generation was shown to be specific since it could be effectively inhibited by active bovine SOD, but not by heat-inactivated enzyme or catalase. Conversely, the xanthine/xanthine oxidase system can merely produce a LBCL peak rapidly but decay instantaneously. To illustrate the application of the proposed method for assessing the O2 --scavenging ability of various food extracts, namely, Prunus mume (A), Lilum lancifolium (B), Creataegus pinnatifida (C), Tremella fuciformis (D), Fortunella margarita (E), and Scutellaria baicalensis (F), we used the following protocol: 12 min after monitoring of LBCL, 1 mg/mL of each of the test compounds was added to the assay system and various degrees of sudden drop of LBCL values were observed, indicating differences in O2 --scavenging abilities exerted by these food extracts that can be visually compared. Consequently, the percentages of inhibition of LBCL versus the concentrations of a test compound can be constructed. It follows that the concentration needed to inhibit 50% of LBCL (IC50) of a test compound can be extrapolated from the curve. Using this approach, we were able to obtain the IC50 values of various compounds to be tested and the order of inhibitory efficiency of the above-mentioned food extracts was ranked, being A > B > C > D > E > F, respectively.

KW - Arginine

KW - Chemiluminescence emitter

KW - Food constituents

KW - Methylglyoxal

KW - Superoxide-scavenging ability

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U2 - 10.1021/jf020799t

DO - 10.1021/jf020799t

M3 - Article

VL - 51

SP - 58

EP - 62

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 1

ER -