Determination of flavone glycosides in pollen of Brassica campestris by HPLC

Objective: To develope a HPLC method for determination of quercetin and kaempferol in the pollen of Brassica campestris. Methods: Quantitation of quercetin and kaempferol was conducted by HPCL, whose conditions were as follows: an Eclipse XDB-C_8 column (150mm× 4.6mm,5μm), a mixture of methanol-0.4% phosphoric acid (50:50) as an eluant with an eluant rate of 1.0 mL·min~(-1), and the detection at 360nm. The sample was initially loaded onto the HPLC in a volume of 10μL,and quantified via the external standard of the peak area. Results: Quercetin and kaempferol were separated well from other ingredients. Quercetin and kaempferol showed a good linear curve in the range of 0.029～0.73μg (r=0.9999) and 0.053～1.315μg (r=0.9999), respectively, and a linear equation Y=3727.7X-6.0 and Y=3932.6X+3.5,respectively. The mean recovery of quercetin and kaempferol was 99.7% with RSD of 1.6% and 99.6% with RSD of 2.0%. Conclusion: This reproducible and reliable HPLC analysis provided a quantitation of flavone glycosides in the pollen.