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Abstract
A simple and reliable method of ultra-performance liquid chromatography coupled with a photodiode array detector (UPLC-PAD) was developed for the quantitative analysis of 10 active nucleosides and nucleobases. The separation was performed on a reversed-phase C18 column with linear gradient elution. This method provided good reproducibility and sensitivity with excellent intra- and inter-day precision and the validated method was successfully used for the determination of the 10 components of interest of F. unibracteata samples. The quantitative data of the fingerprints were analyzed by similarity analysis (SA) and hierarchical clustering analysis (HCA). Results showed that samples from different regions could be successfully divided into two groups in accordance with their locations of origin. A rapid and convenient discriminatory function using UPLC fingerprint combined with HCA was then established to differentiate the F. unibracteata from different growing regions, which could be helpful for quality evaluation and control of F. unibracteata and related medicinal plants in the future.
ACKNOWLEDGMENT
This research was supported by a grant from the National Key Technology R&D foundation of China (Grant number: 2006BAI09B05-3) and The Chinese Pharmaceutical Industry R&D Project (No. 200707007).
Notes
a Collection time means year and month (i.e., 2007.8 means August 2007).
a y, peak area; x, concentration of compound (µg/mL).
b limit of detection (LOD), S/N = 3.
c limit of quantification (LOQ), S/N = 10.
a RSD (%) = (SD/mean) × 100.
b Accuracy (%) = (mean of measured concentration/spiked concentration) × 100.
a Content = mean ± SD (n = 3).
b RSD (%) = (SD/mean) × 100.
c Recovery (%) = (detected amount – original amount)/spiked amount × 100.
a Content = mean ± SD (n = 3).
ND = not detected.