Validated
Method for the Analysis of Frangulins A
and B and Glucofrangulins A and B Using HPLC and UHPLC
Immanuel Rosenthal
Evelyn Wolfram
Samuel Peter
Beat Meier
10.1021/np400736s.s001
https://acs.figshare.com/articles/journal_contribution/Validated_Method_for_the_Analysis_of_Frangulins_A_and_B_and_Glucofrangulins_A_and_B_Using_HPLC_and_UHPLC/2311078
In the present study, robust and
validated HPLC and UHPLC methods
for the quantitative determination of frangulins A and B (<b>3</b> and <b>4</b>) and glucofrangulins A and B (<b>1</b> and <b>2</b>) in the bark of <i>Frangula alnus</i> have been
developed. The HPLC method allowed the separation of the analytes
in 25 min and the UHPLC method in just 13 min. The HPLC method used
an MN Nucleodur C<sub>18</sub> 125 × 4 mm column with 3 μm
particles, while the UHPLC method used a Waters Acquity UPLC BEH C<sub>18</sub>, 100 × 2.1 mm column with 1.7 μm particles. Mobile
phase A consisted of water and 1.25 mL/L phosphoric acid (85%), while
mobile phase B consisted of CH<sub>3</sub>CN/MeOH (20:80). The flow
rates were set to 1 mL/min for the HPLC method and 0.4 mL/min for
the UHPLC method, with the column temperature held at 50 °C and
the detection wavelength being 435 nm for either method. A fractional
factorial design was used to check the robustness of the methods.
The resolution of the analytes was never less than 1.5 when the factors
were varied in the tested range. The conditions for ultrasonic extraction
were optimized by response surface methodology and found to be 68%
CH<sub>3</sub>CN in the extraction solvent, 35 °C extraction
temperature, and a duration of 25 min. The extraction procedure was
determined to be very robust against small deviations of these factors.
2014-03-28 00:00:00
3CN
1.7 μ m particles
BEH
UHPLC method
UPLC
25 min
MN
extraction
CH
3 μ m particles
response surface methodology
HPLC method