摘要
目的建立反相高效液相色谱法测定三妙丸中黄柏生物碱微透析探针体内外回收率,考察回收率影响因素及体内稳定性。方法采用增量法和减量法,用RP-HPLC法测定微透析样品中药根碱、巴马汀和小檗碱的质量浓度,考察流速及质量浓度对探针回收率的影响;探针分别植入颈静脉和膝关节,考察生物碱体内回收率稳定性。结果药根碱、巴马汀和小檗碱的质量浓度在0.04~1.0 mg·L-1内与峰面积呈良好的线性关系（r=0.999 8,n=7）,本方法的专属性、精密度、准确度和稳定性等均符合微透析样品的分析要求。膝关节探针以1.0μL·min-1灌流,药根碱、巴马汀和小檗碱的相对回收率（RR）和损失率（RL）分别为（29.38±2.02）%和（31.80±1.88）%、（30.48±1.93）%和（30.93±2.09）%及（28.95±0.94）%和（30.53±1.74）%。在质量浓度为0.20~0.80 mg·L-1内,RR和RL一致,探针回收率与流速成反比,与质量浓度无关。体内回收率在5 h内保持稳定。结论反透析法校正大鼠血和膝关节的回收率时,所用的微透析方法可用于三妙丸中黄柏生物碱体内药动学研究。
Objective To develop an high performance liquid chromatography coupled with microdialysis sampling method for the determination of in vitro and in vivo microdialysis recovery and influence factors of three alkaloids of Phellodendri Chinensis Cortex in Sanmiao Wan. Methods Gain method and loss method were used for the study. In vitro experiments, the concentrations of the three alkaloids in dialysates were measured by HPLC, and the effects of flow rate and concentration on recovery were determined; the probes for blood and knee joint sampling were implanted within the jugular vein and the knee joint. In vivo experi- ments were performed on rats to measure the stability of the recovery of the three alkaloids in dialysates. Results All calibration curves gave good linearity （ r = 0. 999 8, n = 7 ） over the concentrations range from0. 04- 1.0 mg. L- 1 for all components. The precision and accuracy were evaluated by intra- and inter-day as- says and specificity, stability were all good. Microdialysate contained jateorhizine, palmatine, berberine was passed through the microdialysis probe at a constant flow-rate（ 1.0 μL·min -1 ） in knee joint,the probe rela- tive recovery （RR） and relative loss （RL） were （ 29.38± 2.02 ） % and （ 31.80±1.88 ） %, （ 30. 48 ±1.93 ） % and （ 30. 93 ± 2.09 ） % , （ 28.95 ± 0.94 ） % and （ 30. 53 ± 1.74 ） %, respectively. The probe recovery obtained by gain method was similar to that of loss method over the concentrations range from 0.20- 0. 80 mg. L- 1 for all components. The recoveries of blood and knee joint probes were inversely proportional to the perfusing rate and independent of the concentrations of alkaloids for the microdialysis experiments. The recovery results showed good stabilities in 5 hours in vivo. Conclusions Providing that the relative re- coveries of experiment in blood and knee joint are determined by retrodialysis, microdialysis can be used for the pharmacokinetic study of three alkaloids of Phellodendri Chinensis Cortex in San
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2014年第9期697-705,共9页
Journal of Shenyang Pharmaceutical University
基金
国家自然科学基金资助项目(81274106)
关键词
微透析
回收率
药根碱
巴马汀
小檗碱
血
膝关节
microdialysis
recovery
jateorhizine
palmatine
berberine
blood
knee joint
