The extracts were prepared using 0.2 g of dry leaf matter at 35°C with 4 mL of methanol for HPLC (Neon®) in an ultrasound bath for 30 min. The filtration was performed through a membrane Millipore (Advantec HP020AN−20 μm). Then, chromatographic analysis was performed using a Shimadzu HPLC with an SPD-M20A photodiode matrix detector (λ = 254 nm) and an LC18 column (25 cm × 4.6 mm, 5 μm, Supelcosil™) coupled to a 2 cm LC18 precolumn (Supelguard, Supelco). The mobile phase consisted of Milli-Q water (A) and acetonitrile (B) at a flow rate of 0.6 ml/min. The oven was adjusted to 30°C. The sample injection volume was 20 μL, and the detection was 254 nm, following a method adapted from Morais et al. (2018 (link)). The furanocumarin concentration present in the extracts was calculated from a four data point calibration curve psoralen and bergapten standard, respectively. The concentration was expressed in micrograms of metabolite per gram of leaf matter (μg/g−1 LM). The yield was calculated as the dry weight (g) of the leaves multiplied by the production of the metabolite (μg/g−1).
Supelcosil
Manufactured by Merck Group
Sourced in United States
Supelcosil is a line of high-performance liquid chromatography (HPLC) columns manufactured by Merck Group. The columns are designed for a variety of analytical applications and provide reliable and consistent separation of various chemical compounds.
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Lab products found in correlation
2 protocols using supelcosil
1
HPLC Analysis of Furancoumarin Compounds
2
HPLC Analysis of Monomer Leaching from Dental Adhesives
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The extracts from the adhesives were analyzed by HPLC using an instrument equipped with a photodiode array detector (208 nm), an auto sampler, a pump and a C18 steel column 250 mm in length and 4.6 in diameter, with a particle size of 5 µm (SUPELCOSIL™; Supelco, Bellefonte, PA, USA). The mobile phase was a solution of 80% acetonitrile and 20% water that flowed at a rate of 1 ml/minute. The injection volume was 20 µL. Calibration curves were drawn for each monomer at different concentrations. In this way, standard chromatograms of Bis-GMA, UDMA, HEMA and TEGDMA (Sigma Aldrich, St. Louis, MO, USA) were obtained. The quantity of monomers leaching from the adhesives was obtained by a linear regression analysis of the results from the monomer standard series. A linear calibration equation for the monomers is shown in Table II.
One-way analysis of variance was used to analyze data on the quantity of monomers released by the adhesive materials after different polymerization times. Multiple comparisons were analyzed using the Tukey HSD test at a significance level of p<0.05.
One-way analysis of variance was used to analyze data on the quantity of monomers released by the adhesive materials after different polymerization times. Multiple comparisons were analyzed using the Tukey HSD test at a significance level of p<0.05.
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