The simultaneous separation and determination of six flavonoids and troxerutin in rat urine and chicken plasma by reversed-phase high-performance liquid chromatography with ultraviolet-visible detection.
The method of high-performance liquid chromatography (HPLC) with UV-vis detection was used and validated for the simultaneous determination of six flavonoids (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and troxerutin in rat urine and chicken plasma. Chromatographic separation was performed using a VP-ODS column (150 mm x 4.6 mm, 5.0 microm) maintained at 35.0 degrees C. The mobile phase was a mixture of water, methanol and acetic acid (57:43:1, v/v/v, pH 3.0) at the flow rate of 0.8 mL/min. Six flavonoids and troxerutin were analyzed simultaneously with good separation. On optimum conditions, calibration curves were found to be linear with the ranges of 0.10-70.00 microg/mL (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and 0.50-350.00 microg/mL (troxerutin). The detection limits were 0.010-0.050 microg/mL. The method was validated for accuracy and precision, and it was successfully applied to determine drug concentrations in rat urine and chicken plasma samples from rat and chicken that had been orally administered with six flavonoids and troxerutin.
Yang
Gong-Jun
GJ
College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, 225002, PR China. yanggongjun888@163.com
Liu
Ping
P
Qu
Xi-Long
XL
Xu
Min-Juan
MJ
Qu
Qi-Shu
QS
Wang
Cheng-Yin
CY
Hu
Xiao-Ya
XY
Wang
Zhi-Yue
ZY
D000818:Animals; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005419:Flavonoids / Q000097:blood / Q000302:isolation & purification* / Q000652:urine; D006896:Hydroxyethylrutoside / Q000031:analogs & derivatives* / Q000097:blood / Q000302:isolation & purification / Q000652:urine; D051381:Rats; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
D005419:Flavonoids; D006896:Hydroxyethylrutoside; C005865:troxerutin