ABSTRACT
A new HPLC-UV technique for the separation and analysis of 10 monosaccharides achieved within 13.5 min using 1-phenyl-3-methyl-5-pyrazolone (PMP) as the labelling molecule of the reductive monosaccharides has been established by combining common high performance liquid chromatography-UV and C18 column. The established technique was applied to the quantification of the monosaccharide components in extract of Silybum marianum. The results showed that the tested 10 monosaccharides as PMP derivatives were baseline separated under the HPLC conditions proposed. It was confirmed that Silybum marianum extract was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose with the molar ratio of 0.66:0.84:0.58:1.0:1.6:0.69:2.7:4.8. Quantitative recoveries of the compositional monosaccharides separated from the extract were in the range of 92.4%-104.0%, and the RSD values fell within 0.68%-3.81%. The results demonstrated that the proposed HPLC method was simple, rapid, convenient, and precise, and it was applicable to the analysis of the compositional monosaccharides of Silybum marianum extract.
Subject(s)
Antipyrine , Chemistry , Arabinose , Chromatography, High Pressure Liquid , Methods , Galactose , Glucose , Glucuronic Acid , Hexuronic Acids , Mannose , Silybum marianum , Chemistry , Monosaccharides , Plants, Medicinal , Chemistry , Polysaccharides , Chemistry , Quality Control , Rhamnose , Seeds , Chemistry , Spectrophotometry, Ultraviolet , Methods , XyloseABSTRACT
<p><b>AIM</b>To study the pharmacokinetics of genistein in Beagle dogs.</p><p><b>METHODS</b>Genistein, suspended in 0.5% CMC-Na solution, was orally administered to Beagle dogs at the dose of 5.34 mg.kg-1. At various time intervals, 1.5 mL of blood was drawn from the vein of dogs in their front legs. At the same time, urine and feces were collected. After the collection, the feces were homogenized with physiological saline (to 1 g feces, 10 mL physiological saline were added). The genistein in plasma, urine and homogenized feces was extracted twice by vortexing with 2.0 mL mixture of methyl tert-butyl ether and pentane (8:2). The organic phase was transferred into tubes and evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol and 20 microL of the solution was drawn and detected by high-performance liquid chromatography. The pharmacokinetic parameter was calculated by 3P97 software.</p><p><b>RESULTS</b>The plasma concentration-time curve was fitted to a one-open-compartment model. The peak time was 0.29 h, and the elimination half-life was 0.52 h. After genistein was administered, 10.79% of genistein were excreted from urine and 21.55% from feces within 24 h. It was also found that 13.00% genistein were excreted from urine and 52.46% from feces within 60 h.</p><p><b>CONCLUSION</b>It showed that the speed of absorption and elimination of genistein was high in Beagle dog, and genistein was mainly excreted in the form of parent compound in urine and feces.</p>