Quantitation Of Trans-nerolidol In Rat Plasma By LC-MS/MS And Its Application To Pharmacokinetics Study

Objective:Trans-nerolidol has a wide range of medicinal value,such as antioxidant activity,antibacterial,antitumor,anti ulcer activity,but a trans-nerolidol pharmacokinetic research method has not been established.Therefore,we established a LC-MS/MS method for the determination of trans-nerolidol in rat plasma to provide the basis of trans-nerolidol pharmacokinetic studies.Methods:The optimal chromatographic separation was conducted on a Synergi 4u Fusion-RP C₁₈8 column?50 mm×3.0 mm,4?m? from Phenomenex?MA,USA?.The mobile phase was made of acetonitrile–a 0.1%formic acid and 1 mmol/L ammonium acetate aqueous solution mixture?40:60,v/v? and was delivered at a flow rate of 400?L/min.The total run time was 3 min,and the injection volume was 40?L.The mass spectrometer was conducted in the positive multiple reaction monitoring?MRM? mode via Turbo V electrospray ion source.The ion spray voltage was set to 5.5 kV with a source temperature of 650?.The curtain gas was set to 25 psi.The nebulizer gas?gas 1?was set to 60 psi.The heater gas?gas 2? was set to 65 psi.The optimized MRM mass transitions,the adjusted values of declustering potential and collision energy were:m/z 221.2/135.2,34 V and 15 eV for trans-nerolidol;m/z 254.1/108.1,40V,31eV for sulfamethoxazole?Internal Standard,IS?.Six male Sprague-Dawley rats were given single dose of trans-nerolidol at 50 mg/kg.Blood samples were collected via orbital venous plexus at ten time points after intragastric administration.Simple one-step extraction with acetonitrile to precipitate protein with only 50?L rat plasma was employed to prepare plasma samples.The established method was validated with specificity,linearity,accuracy,precision,extraction recovery,matrix effect and stability in accordance with Chinese Pharmacopoeia 2015 Edition Volume IV,FDA and EMA regulations.The plasma concentration of trans-nerolidol was determined by LC-MS/MS.Sulfamethoxazole was selected as internal standard.The calibration curve was evaluated by plotting the trans-nerolidol peak area/IS peak area?y?versus the trans-nerolidol concentration?x?in rat plasma.The pharmacokinetic parameters of trans-nerolidol were calculated using a non-compartmental model.Results:The calibration curve was linear,with the correlation coefficient being 0.9972,in the linear range of 0.01?g/mL-40?g/mL for trans-nerolidol.The accuracy was in the range of-3.6% to 7.4%,and the intra-and inter-day variations were?7.7%.The mean extraction recovery of all analytes from rat plasma was 91.3-94.3% for trans-nerolidol,and 90.5% for IS.Stability testing showed that trans-nerolidol remained stable during the whole analytical procedure.The present LC-MS/MS method was implemented to support a pharmacokinetic of trans-nerolidol in rats.The plasma trans-nerolidol pharmacokinetic profile and parameters were obtained.Conclusion:A LC-MS/MS method was developed and validated for quantitative analysis of trans-nerolidol in rat plasma.The established method was successfully performed to a pharmacokinetic of trans-nerolidol in rats after intragastric administration.The method was specific,accurate and sensitive.These findings supply evidence for pharmacological action and clinical application of trans-nerolidol.