Amotosalen hydrochloride was purchased from MedChem-Express (> 98%). Coumarin 120 was supplied by Radiant Laser Dyes. Pure water (Fisher Chemical, HPLC gradient grade) and methanol (VWR Chemicals, HPLC gradient grade, ≥ 99.8%) were used as solvents. The HPLC-purified and lyophilized DNA oligomers were purchased from Sigma-Aldrich. A,T-only (5′-(TA) 20 -3′) single strands were employed to form double strands consisting of 40 base pairs. G,C-only (5′-(GC) 4 -3′) single strands were employed to form double strands consisting of 8 base pairs. Longer A,Tstrands were employed because of the lower propensity of A,T-only DNA to form double strands [32] . Solutions of the oligonucleotides were buffered with PBS (Sigma-Aldrich, one tablet dissolved in 200 mL yielded 10 mM phosphate buffer, 2.7 mM potassium chloride, 137 mM sodium chloride, pH 7.4 at 25 °C). Annealing of the oligonucleotide strands in solution was performed 24 h before the measurements. The solution was heated in a water bath up to 95 °C and cooled down to room temperature within several hours. All DNA concentrations refer to base pairs (bp). This is required due to the different strand lengths of A,T-only and G,C-only DNA.
Hplc gradient grade
Manufactured by Avantor
Sourced in United States, Denmark
HPLC gradient grade is a specialized type of high-performance liquid chromatography (HPLC) solvent. It is designed for use in gradient elution HPLC analysis, providing a consistent and reliable mobile phase composition for enhanced separation and detection of analytes.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Mucin, by Merck Group (1 mentions)
Acetic acid, by Avantor (1 mentions)
Nitric acid, by Avantor (1 mentions)
Pepsin, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
Kaempferol 3 o glucoside, by Extrasynthese (1 mentions)
Lipase, by Merck Group (1 mentions)
Bovine bile extract, by Merck Group (1 mentions)
Tyrosol, by Extrasynthese (1 mentions)
Ferulic acid, by Extrasynthese (1 mentions)
Isorhamnetin 3 o rutinoside narcissin, by Extrasynthese (1 mentions)
α amylase from bacillus sp, by Merck Group (1 mentions)
Lc 20at, by Shimadzu (1 mentions)
Sil 20ht autosampler, by Shimadzu (1 mentions)
Milli q system, by Merck Group (1 mentions)
Methanol, by Avantor (1 mentions)
Sodium bicarbonate nahco3, by Merck Group (1 mentions)
Amino acid calibration standard mix, by Merck Group (1 mentions)
6 protocols using hplc gradient grade
1
Oligonucleotide Annealing Protocol for DNA Biophysics
2
Multimethod for Pesticide Extraction
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Stored samples (−20°C) were further cooled to −45°C over night and then homogenized using a knife mill (Grindomix GM 3000, Retsch). An aliquot of the homogenate (10 g) was processed according to the QuEChERS multimethod for pesticides12 using acetonitrile (HPLC gradient grade, VWR) as solvent and QuEChERS tubes I and II (DisQuE 50 mL Tube/AOAC – Acetate and DisQuE 2 mL Tube – AOAC, Waters) for clean‐up. For SDS, DBNS, and docusate extract I (after clean‐up with tube I) was analyzed, and for DMDA, spiroxamine, and trifloxystrobin extract II (after clean‐up with tube I and II). Two replicates of each homogenate were processed and analyzed in separate series on consecutive days. For quality control, fortified samples were processed and analyzed in each series.
3
Antioxidant Capacity Measurement Protocol
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For use as reagents, 6-Hydroxy-2, 5,7,8-tetramethylchromane-2-carboxylic acid (Trolox n 648471, Calbiochem San Diego, CA, USA,), 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS, A 1888, Sigma Aldrich, Taufkirchen, Germany)), 1,1-Diphenyl-2-picrylhydrazyl (DPPH, Sigma Aldrich), phosphate buffer solution (PBS), Ethanol (EtOH), Methanol (MtOH HPLC grade), Trifluoracetic acid, (TFA, Sigma Aldrich), nitric acid and acetic acid (HPLC gradient grade) were obtained from VWR Chemicals (Radnor, PA, USA). All other reagents were of analytical grade and the water was ultra-pure-grade. Phenolic standards: ferulic acid, tyrosol, isorhamnetin 3-O-rutinoside (narcissin) and kaempferol 3-O-glucoside were purchased from Extrasynthese (Genay, France). The in vitro digestion enzymes: α-amylase from Bacillus sp; pepsin, pancreatin, mucin, lipase from pigs; and bovine bile extract were obtained from Sigma-Aldrich (Milan, Italy).
4
Quantification of Total Phenolics
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Total phenolics were determined by using Folin–Ciocalteu reagent (Singleton and Rossi, 1965 ). Twenty milligrams of powdered samples (freeze-dried) was extracted for 10 min with 500 μL of 70% methanol (HPLC-Gradient grade, VWR chemicals) at 70 °C. The mixtures were centrifuged at 3500g for 10 min and the supernatants were collected in separate tubes. The pellets were re-extracted under identical conditions. Supernatants were combined and used for total phenolics assay and for HPLC analysis. For total phenolics assay 20 μL of extract was dissolved into 2 mL of distilled water. 200 μL of dissolved extract was mixed with 1 mL of Folin–Ciocalteu reagent (previously diluted tenfold with distilled water) and kept at 25 °C for 3–8 min; 0.8 mL of sodium bicarbonate (75 g L−1) solution was added to the mixture. After 60 min at 25 °C, absorbance was measured at 765 nm. The results were expressed as gallic acid equivalents.
5
HPLC Analysis of Compound Extraction
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About 100 mg of sample powder were extracted in 10 mL of 70% methanol for 15 minutes (HPLC Gradient grade, VWR chemicals) and filtered using a 0.45 μm PTFE syringe filter. The analysis used a Shimadzu LC-20AT high performance liquid chromatography system with a UV-VIS SPD20A detector and SIL-20HT autosampler made in Japan. Furthermore, 20 μL of the extract were analyzed using a column temperature of 35°C and a flow rate of 0.7 mL/min. A nonpolar C18 column (150 × 4.6 mm) with a 5 μm particle size was used. Eluent A was a 40-minutes gradient program using 1% (v/v) acetic acid in ultrapure water, while eluent B used acetonitrile as follows: 0 to 5 minutes: 10% B, 5 to 15 minutes: 40% B, 15 to 20 minutes: 60% B, 20 to 30 minutes: 90% B, 30 to 40 minutes: 10% B. Subsequently, the absorbance was measured at 272 nm [22 (link)].
6
Defatted Hempseed Meal Amino Acid Analysis
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The hempseed meal used in this study was produced as a by‐product after cold oil processing and supplied by a local Irish farm. Defatted hempseed meal pellets were ground using an electric mill (Lloytron E5012WI, Kitchen Perfected Blender with Mill, Lancshire, UK) and vacuum preserved until further analysis. Sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium chloride (NaCl) and sodium bicarbonate (NaHCO3) were of reagent grade and purchased from Sigma‐Aldrich (St Louis, MO, USA). The amino acid calibration standard mix (analytical standard grade) was purchased from Sigma‐Aldrich (Copenhagen, Denmark). Acetonitrile (high‐performance liquid chromatography (HPLC) gradient grade) and methanol (HPLC gradient grade) were from VWR International (Søborg, Denmark). The ultrapure water used in all the experiments was obtained using a Milli‐Q system (Millipore, MA, USA).
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