Study on the interaction of plasma protein binding rate between edaravone and taurine in human plasma based on HPLC analysis coupled with ultrafiltration technique

Abstract

In this work, two high-performance liquid chromatography (HPLC) assays were developed and validated for the independent determination of edaravone and taurine using 3-methyl-1-p-tolyl-5-pyrazolone and L-glutamine as internal standards. In in vitro experiments, human plasma was separately spiked with a mixture of edaravone and taurine, edaravone or taurine alone. Plasma was precipitated with acetonitrile containing 0.1% formic acid. Ultrafiltration was employed to obtain the unbound ingredients of the two drugs. The factors that might influence the ultrafiltration effiency were elaborately optimized. Plasma supernatant and ultrafiltrate containing taurine were derivated with o-phthalaldehyde and ethanethiol in the presence of 40 mmol/L sodium borate buffer (pH 10.2) at room temperature within 1 min. Chromatographic separations were achieved on an InertSustain C18 column (250 × 4.6 mm, 5 µm). Isocratic 50 mmol/L ammonium acetate-acetonitrile and gradient 50 mmol/L sodium acetate (pH 5.3)-methanol were respectively selected as the mobile phase for the determination of edaravone and taurine. All of the validation data including linearity, extraction recovery, precision, accuracy and stability conformed to the requirements. Results showed that there were no significant alterations in the plasma protein binding rate of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible.

Interaction of Citrinin with Human Serum Albumin

“…Since ultrafiltration is a relatively long process (10 min in our experiments), this technique is not suitable to quantify the binding constant. However, ultrafiltration is highly suitable to clearly demonstrate the complex formation and to investigate different competitive interactions (and in this way to identify the binding site with specific site markers) [ 21 , 22 ]. As Figure 2 A demonstrates, the presence of increasing HSA concentrations (1, 2.5, and 5 μM) led to the significant decrease of CIT in the filtrate.…”

Rat plasma protein binding of kaempferol‐3‐ O ‐rutinoside from Lu’an GuaPian tea and its anti‐inflammatory mechanism for cardiovascular protection

“…Rat plasma protein binding of KR was determined by ultrafiltration (Tang et al., 2015). After blood collection from the abdominal aorta, the SD rats were anesthetized with sodium pentobarbital and sacrificed by cervical dislocation.…”

Pharmacokinetic profiles contribute to the differences in behavioral pharmacology of 071031B enantiomers as novel serotonin and norepinephrine reuptake inhibitors

“…The unbound fraction of S-071031B or R-071031B was measured using ultrafiltration technique as described previously (Tang et al, 2015). Fresh blood was obtained from naive rats by exsanguination via cardiac puncture.…”