Direct and rapid measurement of hydrogen peroxide in human blood using a microfluidic device

Abstract

The levels of hydrogen peroxide ($${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 ) in human blood is of great relevance as it has emerged as an important signalling molecule in a variety of disease states. Fast and reliable measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 levels in the blood, however, continues to remain a challenge. Herein we report an automated method employing a microfluidic device for direct and rapid measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in human blood based on laser-induced fluorescence measurement. Our study delineates the critical factors that affect measurement accuracy—we found blood cells and soluble proteins significantly alter the native $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 levels in the time interval between sample withdrawal and detection. We show that separation of blood cells and subsequent dilution of the plasma with a buffer at a ratio of 1:6 inhibits the above effect, leading to reliable measurements. We demonstrate rapid measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in plasma in the concentration range of 0–49 µM, offering a limit of detection of 0.05 µM, a sensitivity of 0.60 µM−1, and detection time of 15 min; the device is amenable to the real-time measurement of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in the patient’s blood. Using the linear correlation obtained with known quantities of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 , the endogenous $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 concentration in the blood of healthy individuals is found to be in the range of 0.8–6 µM. The availability of this device at the point of care will have relevance in understanding the role of $${\mathrm{H}}_{2}{\mathrm{O}}_{2}$$ H 2 O 2 in health and disease.

Electrochemical Detection of H2O2Using an Activated Glassy Carbon Electrode

“…17 For a healthy person, the H 2 O 2 concentration should be <10 nM in blood and plasma, however, it may increase in the plasma samples collected from the unhealthy persons (from 1-5 μM). 18 Until now many researchers have attempted to detect the H 2 O 2 through various methods such as electrochemical, [19][20][21] chromatography, 7 colorimetric, 22,23 luminescent, 24 and spectrophotometric methods. 25,26 Over the past decades, diverse types of carbon-based electrodes have been developed with various scopes and sensitivity for electroanalytical detection of important analytes.…”

Evidence-based pathogenesis and treatment of ulcerative colitis: A causal role for colonic epithelial hydrogen peroxide

“…Systemic redox homeostasis is provided by HSA and erythrocytes, both of which are highly effective scavengers of H 2 O 2 . Their combined action maintains blood H 2 O 2 at very low levels, in the range of 0.8-6 µM for healthy individuals[ 63 ]. HSA can directly scavenge H 2 O 2 via a reduced surface cysteine thiol (cys34)[ 64 ].…”

Mechanochemical synthesis of an AIE-TICT-ESIPT active orange-emissive chemodosimeter for selective detection of hydrogen peroxide in aqueous media and living cells, and solid-phase quantitation using a smartphone

“…11 To date, many tools have been developed to detect H 2 O 2 , such as electrochemical methods, [12][13][14] spectrophotometry, 15,16 chromatography, 17,18 and so on. [19][20][21] However, many of them are not viable for portable use due to shortcomings like the high cost of equipment, the tedious analytical procedure, or time-consuming detection processes. In addition, some of them can cause tissue or cell damage preventing their use in in vivo detection.…”