Determination of andrographolide and dehydroandrographolide in rabbit plasma by on-line solid phase extraction of high-performance liquid chromatography.
The high-performance liquid chromatography (HPLC) coupled with on-line solid phase extraction (SPE) and ultraviolet (UV) detection was developed for determining andrographolide and dehydroandrographolide in rabbit plasma. Plasma samples (100 microL) were injected directly into a C18 SPE column and the biological matrix was washed out for 6 min using 15% aqueous methanol. By rotation of the switching valve, andrographolide and dehydroandrographolide were eluted in the back-flush mode and transferred to the analytical column by the chromatographic mobile phase consisted of methanol:acetonitrile (ACN):water (50:10:40; v/v). The UV detection was performed at 225 nm. The calibration curves showed excellent linear relationship (R> or =0.9993) over the concentration range of 0.05-5.0 microg mL(-1). The within- and between-day precisions (R.S.D.) of two analytes were in the range of 1.2-6.5% and the accuracies were between 92.0% and 102.1%. Their recoveries were all greater than 94%. The limits of detection were 0.019 microg mL(-1) for andrographolide and 0.022 microg mL(-1) for dehydroandrographolide. This method was successfully applied to the plasma concentration-time curve study after oral administration of Andrographis paniculata Nees extract in rabbit.
Chen
Ligang
L
College of Chemistry, Jilin University, 2699 Qianjin Street, Changchun 130012, PR China.
Yu
Aimin
A
Zhuang
Xiaodong
X
Zhang
Kui
K
Wang
Xiupin
X
Ding
Lan
L
Zhang
Hanqi
H
D035685:Andrographis / Q000737:chemistry*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes / Q000097:blood*; D005260:Female; D008297:Male; D009862:Online Systems; D010936:Plant Extracts / Q000493:pharmacokinetics*; D011817:Rabbits; D015203:Reproducibility of Results; D052616:Solid Phase Extraction
D004224:Diterpenes; D010936:Plant Extracts; C030419:andrographolide; C478098:dehydroandrographolide