Lc solution software version 1

Molecular weight (M_w) of samples was carried out by high-performance size-exclusion chromatography using an HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a LC-20 AD liquid chromatograph, SIL-20A auto sampler, a Yarra 3 µm SEC-2000 column (300 × 7.8 mm; Phenomenex, Torrance, CA, USA) and coupled with a RID-10A refractive index detector (Shimadzu Corporation, Kyoto, Japan). The samples were made according to Dranca et al. [13 (link)]. The LC solution software version 1.21 (Shimadzu Corporation, Kyoto, Japan) was utilized to collect the data.

Honey solutions were prepared following the steps presented in Section 2.3.3 [30 (link)]. The samples were filtered through 0.45 µm PTFE membrane filters and then injected (with a volume of 10 µL) into the HPLC instrument (Schimadzu, Kyoto, Japan) for analysis using an SPD-M-20A diode array detector. The separation was carried out on a Phenomenex Kinetex 2.6 μm Biphenyl 100 Å HPLC Column 150 × 4.6 mm thermostated at 25 °C. Elution was carried out with a solvent system consisting of 0.1% acetic acid in water (solvent A) and acetonitrile (solvent B) as previously described by Palacios et al. [32 (link)] with modifications. The solvent flow rate was of 1 mL·min⁻¹. The determined phenolic compounds were gallic acid, vanillic acid, protocatechuic acid and p-hydroxibenzoic acid at 280 nm, and chlorogenic acid, p-coumaric acid, caffeic acid, rosmarinic acid, myricetin, quercetin, luteolin and kaempherol at 320 nm. The obtained standard calibration curves showed high degrees of linearity (R² > 0.99). Data collection and subsequent processing were performed using the LC solution software version 1.21 (Shimadzu, Kyoto, Japan).

Molecular weight (Mw) was determined by high-performance size-exclusion chromatography using a HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a LC-20 AD liquid chromatograph, SIL-20A auto sampler, a Yarra 3 µm SEC-2000 column (300 × 7.8 mm; Phenomenex, Torrance, CA, USA) and coupled with a RID-10A refractive index detector (Shimadzu, Kyoto, Japan). The samples were made according to Dranca et al. [31 (link)]. Elution was carried out with 0.1 M sodium nitrate solution containing 0.024% sodium azide at a flow rate of 0.5 mL/min and temperature of 25 °C. Pectin solutions (0.3% w/w) were filtered through 0.45 μm membranes before injection. The calibration was performed using pullulan standards (Shoko Science Co., Tokyo, Japan). The LC solution software version 1.21 (Shimadzu Corporation, Kyoto, Japan) was utilized to collect the data.