Developed and validated HPLC method for simultaneous analysis of key flavonoids and phenolcarboxylic acids in the hawthorn-based cardiotonic Forticor®

Abstract

<p>Relatively simple, but very reliable, one-step qualitative/quantitative HPLC analysis of four key flavonoids, and a phenolcarboxylic acid, has been introduced. For substrate/sample preparations, either triple percolations, using 70% ethanol-water (V/V), or/and ultrasonic bath methanol extractions, were carried out to obtain the desired isolates yielding quercetin, isoquercitrin, hyperoside, vitexin and chlorogenic acid. The method is linear, over the studied range of 1.05 <span style="background-color: var(--bs-body-bg); font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align);">– </span><span style="background-color: var(--bs-body-bg); font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align);">210.00, 6.25 </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">–</span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> 250.00, 25.00 – </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">250.00, 7.50 – 150.00 and 1.04 - 10.40 µg/mL for chlorogenic acid, vitexin, hyperoside, isoquercitrin and </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">quercetin, respectively. The correlation coefficient for each of the analytes was greater than 0.999. The </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">intra-day and inter-day precision of the analysis was below 2.00 and 3.00 %, respectively. The accuracy of the analysis is verified by the standard addition method, using three different concentrations of each component in the tested materials, with recovery values obtained in the range of 98.04 </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">–</span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);"> </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">102.47% </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">(RSD ≤ 1.85%). The detection limits were 0.6, 0.5, 0.5, 0.8 and 0.3 µg/mL chlorogenic acid, vitexin, </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">hyperoside, isoquercitrin and quercetine, respectively. The developed method is convenient in the </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">routine control of hawthorn raw materials and pharmaceutical dosage forms made from hawthorn </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">berries or flowerbearing branches. The new HPLC method may be used for the quality control of the </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">commercially used cardiotonic Forticor<sup>®</sup> intended for increasing/strengthening cardiac muscles, in </span><span style="font-weight: var(--bs-body-font-weight); text-align: var(--bs-body-text-align); background-color: var(--bs-body-bg);">order to lower the risks of atherosclerosis, hypertension and congestive heart failure.</span></p>