HPLC separation and determination of dicoumarol and other simple coumarins in sweet clover

Abstract

Dicoumarol is a mycotoxin, that acts as a blood anticoagulant, is formed during the microbial action of molds and fungi in spoiled hay or silage containing high-coumarin plant. A HPLC-DAD method for determination of coumarins, including dicoumarol, coumarin, and 4-hydroxycoumarin was developed. Methanol and acetic acid were used as mobile phase with gradient elution. The simultaneous separation was performed using C18 type of stationary phase. The recoveries were 88.6 – 92.6 %, 91.8 – 95.0 %, and 89.7 – 94.1 % (evaluated for three concentration levels) for dicoumarol, coumarin, and 4-hydroxycoumarin respectively. The parameters of system suitability (repeatability of retention times and peak areas) were determined for evaluation of the method. The method showed a good linearity in the concentration range 0.7 – 100 μg.mL−1for dicoumarol, 0.05 – 100 μg.mL−1for coumarin and 4-hydroxycoumarin with correlation coefficients higher than 0.9885. Extracts of sweet clover herb, hay, and spoiled hay were subjected to HPLC-DAD analysis. The most abundant compound in sweet clover herb and hay extracts was coumarin. In spoiled sweet clover hay extract the 4-hydroxycoumarin was detected in addition. The formation of 4-hydroxycoumarin was also observed in the synchronous fluorescence spectra recorded at the wavelength difference of 90 nm (difference between emission and excitation wavelength).

Synthesis and biological evaluation of esculetin derivatives as potential anti-HBV agents

“…Hydroxyphenol and coumarin compounds, widely found in nature [ 9 ], have good activity against a variety of viruses, such as HBV and coronavirus [ 10 ]. Among them, esculetin is an important coumarin derivative with a common structure of 6,7-dihydroxy-2-H-1-benzopyran-2-one.…”

Design, synthesis and biological evaluation of esculetin derivatives as potential anti-HBV agents

“…Coumarin is a kind of natural compound widely distributed in nature [7]. Among them, esculetin is an important coumarin derivative with a common structure of 6,7-dihydroxy-2-H-1-benzopyran-2-one.…”

An Easy Procedure to Quantify Anticoagulant Rodenticides and Pharmaceutical Active Compounds in Soils

“…Along with a robust extraction method, adequate detection analysis is necessary. ARs and PhACs have been determined mainly by liquid chromatography (LC) with different detectors such as UV [ 32 , 51 ], diode array (DAD) [ 30 , 52 ], DAD with fluorescence detectors (FLD) [ 53 , 54 ] and single quadruple mass spectrometry (MS) detectors [ 28 , 55 ]. However, the preferred option when looking for sensitivity and selectivity at trace level is liquid chromatography-tandem mass spectrometry (LC-MS/MS) [ 7 , 35 , 56 , 57 ], which is the best option for low-level residues that can be expected to be found in soils.…”