KCR in the eluted fraction was determined using a Waters Nova-Pak® C18 reversed-phase column (3.9 × 300 mm) with a 4 µm particle size. The injection volume was 10 µL; the column oven was at room temperature; the mobile phase was methanol/acetonitrile/aqueous formic acid (0.3%), 60/3/37, v/v, at a flow rate of 0.75 mL/min. Detection was made at 313 nm. The results of the analysis were expressed in µg of KCR per mL of plasma.
Nova pak c18 reversed phase column
Manufactured by Waters Corporation
Sourced in United States
The Nova-Pak® C18 reversed-phase column is a chromatographic column used for liquid chromatography. It features a stationary phase of octadecylsilane (C18) bonded to silica particles, allowing for the separation and analysis of a wide range of organic compounds.
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4 protocols using nova pak c18 reversed phase column
1
Quantification of KCR in Plasma
2
Chromatographic Separation of MK, AZD, and SUN
3
Flavonoid Composition Analysis Using UPLC
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Composition of flavonoids was determined using a method described by [22 (link)]. The extracted flavonoids were separated and identified by an Agilent UPLC equipped with a Nova‐Pak C18 reversed‐phase column (3.9 × 150 mm, 5‐μm particle size; both from Waters, Milford, MA, USA). Solvent A used was 0.3% (v/v) HCOOH in H2O, while solvent B used was acetonitrile of HPLC purity grade. The flow rate of the solvents was maintained at 1 mL/min. The gradient profile was as follows: 85% of A at 0 min and 25% of A at 40 min. The mobile phase of the gradient elution was as follows: A, acetonitryl with 0.1% formic acid; and B, 1% aqueous formic acid mixture (pH = 2). Chromatograms were recorded using a UV–Vis detector at λ = 370 nm. The separated compounds were identified based on retention time mapping using a set of standards. The quantity of the following flavonoids was determined using standard solutions (0.001–0.01 μg/mL) of individual compounds (Sigma‐Aldrich, Poznan, Poland). Was determined of seven flavonoids: naringenin, vitaxin, rutin, quercetin, apigenin, kaempferol and lutein.
4
Flavonol Composition Analysis by UPLC
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The percentage composition of fl avonols was determined according to the method described by Kobus et al. (2009) . Extracted fl avonols were separated and identifi ed by Agilent UPLC using a Nova-Pak C18 reversed-phase column (3.9 × 150 mm, 5-μm particle size; both from Waters, Milford, MA, USA). Solvent A was 0.3% (v/v) HCOOH in H 2 O, while solvent B was 100% CH 3 CN. The fl ow rate was maintained at 1 mL/min. The gradient profi le was as follows: 85% of A at 0 min and 25% of A at 40 min. Chromatograms were recorded using a UV-Vis detector at λ = 370 nm. Separated compounds of individual fl avonols were determined as isoquercetin, quercetin, hyperoside and kaempferol and their percentage composition was calculated.
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