Bioanalytical Method Development and Validation of Veratraldehyde and Its Metabolite Veratric Acid in Rat Plasma: An Application for a Pharmacokinetic Study.

A simple, sensitive, and rapid UHPLC-MS/MS method was developed for the simultaneous determination of veratraldehyde and its metabolite veratric acid in rat plasma. Cinnamaldehyde was used as an internal standard (IS) and the one-step protein precipitation method with 0.2% formic acid in acetonitrile (mobile phase B) was used for the sample extraction. Reversed C18 column (YMC-Triart C18 column, 50 mm × 2.0 mm, 1.9 µm) was used for chromatographic separation and was maintained at 30 °C. The total run time was 4.5 min and the electrospray ionization in positive mode was used with the transition m/z 167.07 → 139.00 for veratraldehyde, m/z 183.07 → 139.00 for veratric acid, and m/z 133.00 → 55.00 for IS. The developed method exhibited good linearity (r2 ≥ 0.9977), and the lower limits of quantification ranged from 3 to 10 ng/mL for the two analytes. Intra-day precision and accuracy parameters met the criteria (within ±15%) during the validation. The bioanalytical method was applied for the determination of veratraldehyde and veratric acid in rat plasma after oral and percutaneous administration of 300 and 600 mg/kg veratraldehyde. Using the analytical methods established in this study, we can confirm the absorption and metabolism of veratraldehyde in rats for various routes.

Evaluation of iNSiGHT VET DXA (Dual-Energy X-ray Absorptiometry) for assessing body composition in obese rats fed with high fat diet: a follow-up study of diet induced obesity model for 8 weeks.

We examined the precision, accuracy, and capability of detecting changes of Dual-Energy X-ray Absorptiometry (DXA) for the measurements of total-body weight (TBW), total-body fat weight (TBFW), and total-body lean weight (TBLW) in an 8-week follow-up study of rats. Twenty male rats (4-week) were divided into 2 diet groups. For 8 weeks, we measured body composition (TBW, TBFW, TBLW) by DXA and TBW by an electronic scale once a week. In week 8, we measured body composition 5 times by DXA and TBFW by dissecting experiment (EXP) of euthanized rats (12-week). Total-body fat ratio (TBFR) was defined as TBFW/(TBFW+TBLW). The precision of DXA was evaluated by measuring the coefficient of variation (CV) and accuracy was evaluated by comparing DXA-derived data with EXP data. The capability of detecting changes of DXA in follow-up study was verified by analyzing the trend of DXA-derived values over the 8 weeks. For TBW, TBFW, TBLW of DXA, CVs were 0.02 ± 0.01, 0.10 ± 0.05, 0.03 ± 0.02 and errors were - 6.996 ± 3.429 (r = 0.999), + 14.729 ± 3.663 (r = 0.982), - 21.725 ± 4.223 (r = 0.991), respectively. Prediction models were [EXP TBW = - 31.767 + 1.085 (DXA TBW), R2 = 0.998, root mean square error (RMSE) = 1.842] and [EXP TBFR = - 0.056 + 1.177 (DXA TBFR), R2 = 0.948, RMSE = 0.007]. Over 8 weeks, DXA TBW and DXA TBLW steadily increased, DXA TBFW steadily increased followed by saturation or declination, difference of DXA TBFW between 2 diet groups steadily increased. In conclusion, our study verified that DXA (iNSiGHT VET DXA, OsteoSys, Korea) is accurate and precise enough to measure body composition of rats. Additionally, we confirmed the possibility that DXA could be used for the long-term follow-up studies.