三叶青地上部分抗炎提取物固体纳米晶生物利用度及药动学研究

Study on the bioavailability and pharmacokinetics of solid nanocrystals of anti-inflammatory extract from the aerial part of Tetrastigma hemsleyanum Diels et Gilg

目的:建立同时测定大鼠血浆中三叶青地上部分抗炎提取物(THAA)9个效应组分的超高效液相色谱-质谱联用技术(UPLC-MS/MS)定量分析方法,评价THAA固体纳米晶血药浓度、药动学特性及口服生物利用度。方法:采用UPLC-MS/MS,CORTECS^(TM) UPLC-C_(18)色谱柱(2.1 mm×100 mm,1.6μm),流动相为乙腈(A)-水(含0.1%甲酸,5%甲醇)(B),梯度洗脱,建立同时测定大鼠血浆中THAA固体纳米晶主要效应组分新绿原酸、绿原酸、荭草苷、异荭草苷、异荭草苷-2′-O-鼠李糖苷、牡荆苷、牡荆苷2′-O-鼠李糖苷、异牡荆苷、异牡荆苷2′-O-鼠李糖苷血药浓度定量分析方法,考察比较大鼠分别灌胃给药THAA固体纳米晶、THAA原料、THAA普通颗粒剂后的血药浓度、药动学参数及口服生物利用度。结果:与灌胃THAA原料或普通颗粒剂相比,灌胃THAA固体纳米晶后,9个效应组分的达峰时间t_(max)显著缩短;半衰期t_(1/2)分别是原料的1.06~5.36倍,颗粒剂的1.16~5.31倍;达峰浓度C_(max)分别是原料的1.74~2.69倍,颗粒剂的1.61~2.45倍;AUC_(0-∞)分别是原料的1.32~2.66倍,颗粒剂的1.18~2.52倍;相对生物利用度比原料提高了131.93%~266.06%,比颗粒剂提高了117.86%~251.55%,且均呈显著性差异(P<0.05)。结论:构建的THAA固体纳米晶使THAA效应组分群起效更快,血药浓度水平显著增加、维持时间更长,改善并提高了THAA效应组分群的口服生物利用度。

OBJECTIVE To establish a UPLC-MS/MS quantitative analysis method for the simultaneous determination of 9 effect components of anti-inflammatory extract from the aerial part of Tetrastigma hemsleyanum(THAA)in plasma of rat,and to evaluate the improvement of THAA solid nanocrystal plasma concentration,pharmacokinetic properties and oral bioavailability.METHODS UPLC-MS/MS technology was used,CORTECS^(TM) UPLC-C_(18)column(2.1 mm×100 mm,1.6μm),and the mobile phase was acetonitrile(A)-water(containing 0.1%formic acid,5%methanol)(B),gradient elution was established to simultaneously determine the main effect components of THAA solid nanocrystals in rat plasma,neochlorogenic acid,chlorogenic acid,orientin,isoorientin,isoorientin quantitative analysis method for plasma concentration of glycoside-2′-O-rhamnoside,vitexin,vitexin 2′-O-rhamnoside,isovitexin,and isovitexin 2′-O-rhamnoside,to investigate and compare the plasma concentration,pharmacokinetic parameters and oral bioavailability of rats after intragastric administration of THAA solid nanocrystals,THAA raw materials,and THAA ordinary granules.RESULTS Compared with the raw materials of intragastric THAA or ordinary granules,the peak time t_(max)of the 9 effect components of intragastric THAA solid nanocrystals was significantly shortened;the t_(1/2) was 1.06-5.36 times of the raw materials,which was 1.16-5.31 times of the ordinary granules.The peak concentration C_(max) was 1.74 to 2.69 times of the raw material,1.61 to 2.45 times of the ordinary granule;AUC_(0-∞)was 1.32 to 2.66 times of the raw material,which was 1.18-2.52 times of the ordinary granule.The relative bioavailability was 131.93%-266.06%higher than THAA,and 117.86%-251.55%higher than the granules of THAA.And there were significant differences(all P<0.05).CONCLUSIONS The THAA effect component group took effect faster,the blood concentration level was significantly increased,the maintenance time was longer,and the oral bioavailability of the THAA effect component group was improved of the constructed THAA solid nanocrystals.