Citation: WANG Dan , LIU Chun-Sheng , LIU Yan-Xia , AN Zhuo-Ling , LI Peng-Fen , LIU Li-Hong , SHI Li-Fu , HU Jin-Hong . Calibration of Ferulic Acid Linear Probe in Rat Skin by Microdialysis-High Performance Liquid Chromatography On-line Technique. Chinese Journal of Analytical Chemistry, 2012, (01): 164-168. doi: 10.3724/SP.J.1096.2012.10344 [复制]
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2 μL/min,10孔自动进样阀间隔为8 min,在以Hypersil-C18(250 mm×4.6 mm,5 μm)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.