Evaluation of sample preparation and chromatographic separation for the parallel determination of taurine and edaravone in rat tissues using HILIC-MS/MS

Abstract

The quantitative analysis of taurine and edaravone in biological sample is critical in pharmaceutical studies. Although each of them can be individually analyzed by different approaches, concurrent quantification is still a highly challenging task with respect to their great polarity variation and the complex composition of tissue sample. In the present study, to simultaneously determine taurine and edaravone in rat tissue, the sample preparation and chromatographic separation conditions were evaluated and discussed in detail. As for the sample preparation, four kinds of solvent and the volume ratio of the optimal solvent to biological sample were both tested and evaluated based on the chromatographic profile, extraction recovery, and matrix effect (ME). The chromatographic separation was performed in a reverse phase (RP) and two hydrophilic interaction liquid chromatography (HILIC) modes, and the corresponding separation efficiencies were assessed using chromatographic parameters like half-width (W 1/2 ), tailing factor (f t), theoretical plates number (N), and ME. Furthermore, adopted composition of two mobile phase systems and the concentrations of the additives in the optimum buffer system were also investigated on an Atlantis HILIC silica column according to the resultant chromatographic profiles and peak areas of the analytes. The optimal results were obtained when the biological samples were deproteined by 4-fold volume of methanol/acetonitrile (1:3, v/v) and separated on a HILIC column with a gradient elution of acetonitrile/water containing 0.2 % formic acid and 10 mM ammonium formate. The proposed approach was validated and successfully applied to the parallel determination of the tissue distribution of edaravone and taurine in rat tissues.

Pre-Clinical Pharmacokinetic Characterization, Tissue Distribution, and Excretion Studies of Novel Edaravone Oral Prodrug, TEJ-1704

“…The recovery cumulative excretion of edaravone after intravenous administration of 1 mg/kg in urine, bile, and feces were 0.30, 0.48, and 0.017%, respectively. According to Li et al [ 7 ], the peak concentration of edaravone in rat organ tissues (kidney, heart, brain, spleen, and liver) occurred at 30 min after the intravenous administration of 1 mg/kg edaravone and the highest concentrations of edaravone were detected in the kidney. In beagles, Shao et al [ 10 ] reported edaravone half-life of about 9 h and AUC 0–24h of about 26,694 ng/mL·h, 42,090 ng/mL·h, and 55,811 ng/mL·h after intravenous administration of 2.4, 4.8, and 9.6 mg/kg edaravone, respectively.…”

“…The recovery cumulative excretion of edaravone after intravenous administration of 1 mg/kg in urine, bile, and feces were 0.30, 0.48, and 0.017%, respectively. According to Li et al [7], the peak concentration of edaravone in rat organ tissues (kidney, heart, brain, Comparable literature data showed that the half-life of elimination and the AUC after intravenous injection of 3.75 mg/kg edaravone in rats were 0.61 h and 8.76 µg/mL•h [5]. The recovery cumulative excretion of edaravone after intravenous administration of 1 mg/kg in urine, bile, and feces were 0.30, 0.48, and 0.017%, respectively.…”

“…To evaluate a PK of edaravone after administration of TEJ-1704, a sensitive quantitation method for edaravone in rat and beagle dog matrices is needed. The bioanalysis method in HPLC and LC-MS/MS have been reported in rat plasma [ 5 , 6 ], organs including kidney, heart, brain, spleen, liver [ 7 ], bile, urine, and feces [ 8 ]. Aside from Li et al [ 7 ], all the other methods had lower sensitivity with the lower limit of quantification (LLOQ) of 100 ng/mL but required rather large volume of matrices (100 μL).…”

“…The bioanalysis method in HPLC and LC-MS/MS have been reported in rat plasma [ 5 , 6 ], organs including kidney, heart, brain, spleen, liver [ 7 ], bile, urine, and feces [ 8 ]. Aside from Li et al [ 7 ], all the other methods had lower sensitivity with the lower limit of quantification (LLOQ) of 100 ng/mL but required rather large volume of matrices (100 μL). Although Li et al have lowered LLOQ drastically to 2 ng/mL, it was only validated in 5 organ matrices.…”

Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis

“…Metformin and other biguanides (phenformin, buformin) were quantified by HILIC–ESI‐MS/MS in both live and postmortem forensic whole blood using a SeQuant ZIC HILIC column . Similarly, taurine and edaravone, a new combined therapy with commercial potential for treatment of stroke, were also determined by HILIC–ESI‐MS/MS using Atlantis HILIC silica column in rat tissues (kidney, heart, brain, spleen, and liver) simultaneously . Analysis of 2‐oxothiazolidine‐4‐carboxylic acid (OTZ), a cysteine prodrug, in simulated aqueous humour was developed according to FDA guidelines and finally gained well accurate and precise with LOD and LLOQ of 200 and 100 ng/mL.…”