Abstract
In this paper, the interaction of hyaluronic acid (HA) with crystal violet (CV) was investigated carefully by linear sweep voltammetry on the dropping mercury working electrode (DME). In pH 5.0 Britton-Robinson (B-R) buffer solution, CV has a sensitive, well-defined second order derivative linear sweep voltammetric reductive wave at –0.85 V (vs. SCE). After adding a certain amount of HA into CV solution, the reductive peak current decreased without any shift of reductive peak potential. Based on the difference in the reductive peak current, a new voltammetric method for the detection of HA was established. The reaction conditions and the electrochemical determination were studied and optimized. Under the optimized conditions, the decrease of peak current showed a good linear relationship with the HA concentration in the range from 10.0 to 40.0 mg/L. The linear regression equation was got as ∆ip″(nA)= 84.07 C–527.86 (mg/L) (n=8, γ=0.997) and the detection limit was calculated as 2.65 mg/L (3σ). This new established method was further used to HA determination in the synthetic samples with satisfactory results and good recovery. The stoichiometry of CV-HA complex was calculated and the binding mechanism was also discussed by the electrochemical data.
Highlights
As a member of naturally occurring glycosaminoglycan, hyaluronic acid (HA) is consisted of alternating disaccharide units of β-(1-3)-N-acetyl-D-glucoscmine and β-(1-4)-D-glucuronic acid [1]
Curve 2 was that of crystal violet (CV) in pH 5.0 B-R buffer solution, a well-defined voltammetric reductive peak at -0.85 V appeared, which was due to the electrochemical reduction of CV on the mercury electrode
When HA was added into CV solution, the reductive peak current decreased without the shift of the peak potential, which demonstated that CV interacted with HA and a non-electrochemical active supramolecular biocomplex was formed in the solution
Summary
As a member of naturally occurring glycosaminoglycan, hyaluronic acid (HA) is consisted of alternating disaccharide units of β-(1-3)-N-acetyl-D-glucoscmine and β-(1-4)-D-glucuronic acid [1]. To establish a reliable, accurate quantitative determination method of HA is very important in the field of bio-analytical chemistry. The analytical methods for the determination of HA main contains immunochemistry [5] enzyme-linked immunosorbent assay [6], electrophoresis [7] spectrophotometry [8], high-performance liquid chromatography [9], resonance rayleigh scattering technique [10, 11] etc. Owing to the advantages of lower detection limit and wider linear range, electrochemical methods have been successfully used for the determination of different kinds of biomolecules such as DNA [12,13,14], protein [15, 16], heparin [17, 18] and chondroitin sulfate [19,]. A small amount of sample is needed because the electrochemical reaction is often taken place on the interface of the electrode and the solution
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