Ethanolic extract of propolis investigated in the present study was analyzed by the method described by Gültekin-Özgüven et al., 2015, using a Shimadzu 20A series ultra-fast liquid chromatograph (UFLC, Shimadzu Corporation, Kyoto, Japan) coupled to MS detector with electrospray ion source (ESI) and a triple quadrupole analyzer (API-3200 QTRAP, AB Sciex, Framingham, MA, USA). Identification and quantification of phenolic acid composition were performed using standard phenolic acids, namely, p-Hydroxybenzoic acid, gallic acid, vanillic acid, syringic acid, sinapic acid, caffeic acid, trans-cinnamic acid, ferulic acid, p-coumaric acid, and chlorogenic acid, by comparison of retention time and peak area of the used standard and compounds detected in EEP. In addition, rutin was also used. Separation was performed on an Inertsustain C18 column (150 mm × 4.6 mm, 3 μm) with a guard column (4.0 × 10 mm × 2) using a gradient of mobile phase A and B (7.5 mM formic acid and acetonitrile). An increasing gradient of B starting from 5% up to 95% in 20 min was used. The column temperature was set to 40 °C. A volume of injection of 20 μL for each standard mixture was used with a flow rate of 0.5 mL min−1. [32 (link)]
20a series ultra fast liquid chromatograph uflc
Manufactured by Shimadzu
Sourced in Japan
The 20A series ultra-fast liquid chromatograph (UFLC) is a high-performance liquid chromatography (HPLC) system designed for rapid and efficient separation of chemical compounds. The core function of the 20A series UFLC is to provide fast, sensitive, and accurate analysis of a wide range of samples.
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2 protocols using 20a series ultra fast liquid chromatograph uflc
1
Ethanolic Propolis Extract Analysis by UFLC-MS
2
Phenolic Compounds Extraction and UFLC Analysis
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The phenolic extracts were submitted to a liquid-liquid extraction with diethyl ether (2 x 20 mL) and ethyl acetate (2 x 20 mL). Anhydrous sodium sulphate was added to the combined organic phases and the extracts were filtrated through Whatman No. 4 paper, evaporated to dryness and re-dissolved in methanol:water (20:80, v/v). The extracts (1 ml) were filtered through a 0.22 µm disposable LC filter disk. Phenolic acids determination was performed using a Shimadzu 20A series ultra-fast liquid chromatograph (UFLC, Shimadzu, equipment described above) as previously described by Reis et al. (2013) . Detection was carried out in a photodiode array detector (PDA), using 280 nm as the preferred wavelength. The phenolic compounds were quantified by comparison of the area of their peaks recorded at 280 nm with calibration curves obtained from commercial standards of each compound. The results were expressed in µg per 100 g of dry weight.
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