微透析与高效液相色谱在线联用技术校正大鼠皮下阿魏酸线性探针

Calibration of Ferulic Acid Linear Probe in Rat Skin by Microdialysis High Performance Liquid Chromatography On-line Technique

采用微透析与高效液相在线联用技术(MD-HPLC on-line)建立并验证大鼠皮下阿魏酸微透析线性探针体内外校正方法。以磷酸盐缓冲溶液为灌流液,微透析灌注液流速为2μL/min,10孔自动进样阀间隔为8 min,在以Hypersil-C18(250 mm×4.6 mm,5μm)为色谱柱,流动相为甲醇-水(含0.5%乙酸)(35∶65,V/V),流速1 mL/min,检测波长314 nm的HPLC色谱条件下进行在线检测,阿魏酸在0.1～80 mg/L范围内回归方程线性关系良好,A=159044C-2607(r=1),阿魏酸日内精密度RSD分别为0.8%,0.3%和0.5%(n=5);日间精密度RSD分别为0.2%,0.3%和0.4%(n=5),重现性与稳定性RSD分别为0.7%和1.1%(n=5)。增量法和减量法测定阿魏酸线性探针体内外回收率分别为47.23%±0.94%和20.37%±1.37%,阿魏酸适宜进行微透析实验。应用MD-HPLC on-line对中药当归阿魏酸进行测定,使取样、进样和分析监测同时进行。

Microdialysis（MD）-HPLC on-line technique was develop to validate the calibration of ferulic acid linear probe in rat skin,by using phosphate buffer（pH 7.4） as perforates and at perfusion flow rate of 2 μL/min with 8-min intervals.The collective samples were separated by Hypersil-C18 column（250 mm×4.6 mm,5 μm） with mobile phase of methanol-water（0.5% acetic acid）（35：65,V/V） at a flow rate of 1 mL/min and detected at wavelength of 314 nm.The linearity was established for the range of concentrations of 0.1-80 mg/L with a coefficient of determination（r） of 1.The Intra-precision RSDs were 0.8%,0.3%,0.5%（n=5）;inter precision RSDs were 0.2%,0.3%,0.4%（n=5）,and reproducibility and stability RSD were 0.7%,1.1%（n=5）.In vitro and in vivo recovery was calculated by Loss and Gain respectively,and the results were 47.23%±0.94% and 20.37%±1.37%,respectively.We can conclude that microdialysis sampling can be used for the calibration study of ferulic acid.The Chinese angelica ferulic acid was determined by MD-HPLC on-line technique,so that the sampling,injection and analysis can be implemented at the same time.