Catechin, aurantio-obtusin, and chicoric acid were identified and quantified in the extracts of Ulmus pumila L., Cassia tora L., and Taraxacum officinale, and in AF-343 using HPLC–UV (Agilent 1200 series, Agilent, USA; or SHIMADZU Prominence, Shimazu, Japan). Chromatographic separation was performed using various gradients or isocratic elution systems under different mobile phases and wavelengths to establish the optimal separation conditions for each chemical (Figures S1–S3 ). The column used was a Capcell-Pak UG120 C18 column (5 µm, 4.6 × 250 mm, Shiseido, Japan) with a flow rate of 1.0 mL/min. The sample injection volume was 20 µL.
Capcellpak ug120 c18 column
Manufactured by Shiseido
Sourced in Japan
The Capcellpak UG120 C18 column is a laboratory equipment used for high-performance liquid chromatography (HPLC) and ultrahigh-performance liquid chromatography (UHPLC) applications. It is designed to provide efficient separation and analysis of a wide range of compounds.
Automatically generated - may contain errors
Lab products found in correlation
Agilent 1200 series, by Agilent Technologies (1 mentions)
Alliance 2695 hplc system, by Waters Corporation (1 mentions)
Waters 2996 pda detector, by Waters Corporation (1 mentions)
Empower 2 chromatography software, by Waters Corporation (1 mentions)
Agilent 1100 series hplc value system, by Agilent Technologies (1 mentions)
Whatman, by Cytiva (1 mentions)
5 protocols using capcellpak ug120 c18 column
1
Quantification of Bioactive Compounds in Medicinal Plants
2
HPLC Analysis of CME and CPC Fractions
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CME and CPC peak fractions were analyzed using a Waters Alliance 2695 HPLC system coupled with a Waters 2996 PDA detector (Waters, Milford, MA, USA) with a Capcellpak UG120 C18 column (4.6 × 250 mm, 5 μm, Shiseido, Tokyo, Japan). Acetonitrile (0.1% formic acid, solvent A) and water (0.1% formic acid, solvent B) were eluted in a gradient mode: 0–20 min, 10–40% A; 20–25 min, 40–55% A; 25–30 min, 55–95% A; and 40 min, 95% A. The flow rate was 1 mL/min, and the injection volume was 10 μL. The diode array detector measured the UV spectrum over a range of 200–450 nm, and the chromatogram of the effluents was recorded at 254 nm. The system was controlled using Waters Empower™ 2 Chromatography Software.
3
HPLC-DAD Analysis of Bioactive Compounds
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HPLC-DAD analysis was carried out on a Shiseido (Tokyo, Japan) Capcell Pak UG120 C18 column (4.6 × 250 mm, 5 µm); the column temperature was set to 35 °C. The mobile phase consisted of (A) acetonitrile and (B) water at a flow rate of 0.8 mL/min, with gradient elution as follows: 0-5 min, 15% A; 40 min, 46% A; 60 min, 55% A; 70 min, 60% A (v/v); 75 min, 40% A; 76-90 min, 15% A (v/v). Details are explained in the section on method validation in Supplementary Materials .
4
HPLC Profiling of Crude Extract
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The crude extract and its CPC peak fraction were profiled by HPLC equipped with a Capcellpak UG120 C18 column (4.6 × 250 mm, 5 μm, Shiseido, Tokyo, Japan). The mobile phase was composed of acetonitrile containing 0.1% formic acid (A) and water containing 0.1% formic acid (B). The gradient elution conditions were as follows: 0–10 min, 10–50% A; 10–45 min, 50–95% A; and 50 min, 95% A. The column temperature was 40 °C, the mobile phase flow rate was 1 mL/min, the detection wavelength was 230 and 254 nm and the injection volume was 10 µL.
5
Quantitative Analysis of Nucleotides in Milk
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Laboratory instruments were used for general quantitative analysis. Whatman No.1
filter paper (Maidstone, UK) was used for filtration and Macherey-Nagel
Chromabond SB 6 mL/1,000 mg column (Duren, Germany) was used for SPE. The vacuum
manifold used was from Supelco (Maidstone, UK). To filter the final extracted
solution, a 0.22-μm nylon filter (Adventec, Japan) was used. The Agilent
1100 Series HPLC Value System (Agilent, Santa Clara, CA, USA) and the Agilent
1200 Series DAD HPLC System (Santa Clara, CA, USA) were used. The analytical
conditions of the instrument (HPLC-DAD) were optimized prior to initial use. To
effectively detect nucleotides in milk powder and human milk, the sensitivity
was analyzed using DAD, and the Capcellpak UG 120 C18 column
(4.6×250 mm, 5 cm, Shiseido, Japan) was used for separation of
nucleotides. Five optimum separation conditions were used. The mobile phase used
was KH2PO4 (10 mM, pH 5.6), flow rate was 0.6 mL/min,
column temperature was 40°C, total analysis time was 25 min, and
injection volume was 10 μL.
filter paper (Maidstone, UK) was used for filtration and Macherey-Nagel
Chromabond SB 6 mL/1,000 mg column (Duren, Germany) was used for SPE. The vacuum
manifold used was from Supelco (Maidstone, UK). To filter the final extracted
solution, a 0.22-μm nylon filter (Adventec, Japan) was used. The Agilent
1100 Series HPLC Value System (Agilent, Santa Clara, CA, USA) and the Agilent
1200 Series DAD HPLC System (Santa Clara, CA, USA) were used. The analytical
conditions of the instrument (HPLC-DAD) were optimized prior to initial use. To
effectively detect nucleotides in milk powder and human milk, the sensitivity
was analyzed using DAD, and the Capcellpak UG 120 C18 column
(4.6×250 mm, 5 cm, Shiseido, Japan) was used for separation of
nucleotides. Five optimum separation conditions were used. The mobile phase used
was KH2PO4 (10 mM, pH 5.6), flow rate was 0.6 mL/min,
column temperature was 40°C, total analysis time was 25 min, and
injection volume was 10 μL.
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