Applicability of Phenolic Profile Analysis Method Developed with RP-HPLC-PDA to some Bee Product

Abstract

In this work, a reverse phase high-performance liquid chromatographic method (RP-HPLC) with photo diode array (PDA) at 4 different wavelengths (250, 280, 320 and 360) for the determination of some polyphenols in some bee products (honey, pollen, and propolis) is developed. The analyses were carried out on C18 column (250 mm x 4.6 mm, 5 µm; GL Sciences), and the gradient program was used with a mobile phase A reservoir with 70% acetonitrile and the B reservoir with 2% acetic acid. The HPLC method was founded that limit of detection was in the range 0.022-0.0908 µg/mL; the limit of quantification was in the range 0.074-0.3027 µg/mL, all calibration curves were linear R 2 >0.995 within the range, the recovery range was 91.43-111.37% for 10 ppm and 98.44-101.68% for 40 ppm and relative error levels 0.0330-0.0290 respectively. The developed method was applied to some bee products available on the Turkey market. The study aimed at phenolic profiles of the bee products extracts were revealed by using 25 phenolic standards. The proposed method was optimized, quickly, and simple validated by evaluating the linear range, the limits of detection and quantification, the accuracy, the precious, the repeatability, and recoveries suitable for the phenolic analysis. It is concluded that evaluated and the quantitative determination of the bee products can be made quickly and reliably with the optimized method.

In Vitro Determination of Nitric Oxide Synthase Inhibition, Antioxidant Capacity and Phenolic Content of Various Natural Products (Bee and Herbal Products)

“…The solvents were first evaporated in a rotary evaporator (IKA®‐Werke RV 05 Basic) at 40 °C, after which 10 mL of pH 2 distilled water was added, vortexed, and extracted with diethyl ether and ethyl acetate. After all the solvents had been removed, the residue remaining in the flasks was dissolved with 2 ml of absolute methanol, filtered with a 0.45 μm filter (Isolab 094.01.003), and finally injected [21] …”

“…The solvent program began with a linear gradient starting at 95 % A for 3 minutes, followed by a decrease to 80 % A at 10 minutes, 60 % A at 20 minutes, 20 % A at 30 minutes, and finally returning to 95 % A at 50 minutes. A standard chromatogram was generated for 19 phenolic standards, including gallic acid, protocatechuic acid, p ‐OH benzoic acid, catechin, caffeic acid, syringic acid, epicatechin, p ‐coumaric acid, ferulic acid, rutin, myricetin, resveratrol, daidzein, lutein, t ‐cinnamic acid, hesperidin, chrysin, pinocembrin, and CAPE (caffeic acid phenethyl ester) using HPLC with UV detection and the results were expressed as μg/g sample [21,59–62] …”

“…After all the solvents had been removed, the residue remaining in the flasks was dissolved with 2 ml of absolute methanol, filtered with a 0.45 μm filter (Isolab 094.01.003), and finally injected. [21] Each specimen was introduced into the High-Performance Liquid Chromatography (HPLC) system (EliteLaChrom Hitachi, Japan) using a reverse-phase C18 column (150 mm x 4.6 mm, 5 μm; Fortis) with detection at 280 nm. The mobile phase consisted of (A) 2 % acetic acid (Sigma Aldrich.…”

“…A standard chromatogram was generated for 19 phenolic standards, including gallic acid, protocatechuic acid, p-OH benzoic acid, catechin, caffeic acid, syringic acid, epicatechin, p-coumaric acid, ferulic acid, rutin, myricetin, resveratrol, daidzein, lutein, t-cinnamic acid, hesperidin, chrysin, pinocembrin, and CAPE (caffeic acid phenethyl ester) using HPLC with UV detection and the results were expressed as μg/g sample. [21,[59][60][61][62] Total Phenolic Content (TPC) Folin-Ciocalteu's assay was used to measure the extracts' TPC. [21] First, 680 μL extract and 400 μL of 0.2 N Folin-Ciocalteu's reagent (Sigma Aldrich, F9252) were mixed.…”

“…[21,[59][60][61][62] Total Phenolic Content (TPC) Folin-Ciocalteu's assay was used to measure the extracts' TPC. [21] First, 680 μL extract and 400 μL of 0.2 N Folin-Ciocalteu's reagent (Sigma Aldrich, F9252) were mixed. Following 3 min of incubation, 400 μL of 10 % sodium carbonate was added, and the mixture was incubated for 2 h at room temperature.…”