HPLC-MS/MS测定大鼠血浆及组织中盐酸戊乙奎醚浓度

Objective To develop a method for detecting penehyclidine hydrochloride (PH) in mouse plasma and tissues using HPLC-MS/MS. Methods The plasma and tissue samples were extracted with petroleum ether: ethyl ether (70: 30). The isolation of PH was achieved on an Allure C18 (50×2.1 mm, 5μm) column, with methanol/5 mmol/L ammonium acetate/triethylamine=90/10/0.05 (adjusted pH5.8 by formic acid as mobile phase. Benzhydramine was used as the internal standard. Results The calibration curves were linear over the range of 0.39-200 ng/mL for plasma and 0.391-100 ng/mL for tissues. The extraction recovered 67.32%-70.80% of PH in plasma, 84.99%-89.27% of PH in lung tissues, and 76.86%-81.98% of PH in brain tissues. The HPLC detected 97.66%-99.97% of PH in extracted samples of plasma, 96.55%-101.65% extracted samples of lung and 95.18%-100.21% of extracted samples of brain. The within-day RSD were 1.94%-2.93%, 1.73%-3.70% and 2.35%-2.79% for plasma, lung, and brain, respectively. The between-day RSD were 3.00%-3.85% 2.86%-3.94% and 2.77%-5.00% for plasma, lung, and brain, respectively. Both the long-term and freeze-thaw stabilities were acceptable. The long-term RSD were 1.52%-5.26%, 1.88%-2.93%, and 2.22%-3.76% for plasma, lung, and brain, respectively. The freeze-thaw RSD were 0.38%-3.55%, 2.79%-9.60%, and 1.35%-2.29% for plasma, lung, and brain, respectively. Conclusion The assay is simple and accurate, with good reproducibility, and can satisfy to the needs of pharmacokinetics and relative bioavailability studies on penehyclidine hydrochloride.