High performance liquid chromatography (HPLC) was performed on a Dionex Chromelon TM chromatography data system with P580 and UVD100 detectors (Thermo Fisher Scientific Inc., Waltham, MA, USA). Chromatographic separation was performed on a Waters Sunfire C18 column (250 × 4.6 mm, 5‐μm particle size). Elution was performed with a methanol/acetonitrile (3:1) gradient containing 1% formic acid. The gradient was linearly increased from 10% to 90% methanol over 35 min. The injection volume was 10 μl and the flow rate was 1 ml/min.
Dionex chromelon tm
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Dionex Chromelon TM is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative separations. It offers precise control of flow rates, pressures, and temperature, providing efficient and reliable chromatographic separations.
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5 protocols using dionex chromelon tm
1
HPLC Analysis of Compounds
2
Quantification of Apigenin in Plant Extract
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The plant extract was prepared in 50% methanol at the concentration of 2 mg/mL. Serial dilutions (2.5, 25, 125, 250, 500, and 1000 µg/mL) of standard compounds (apigenin) were prepared in methanol. High-performance liquid chromatography (HPLC) was performed on a Dionex Chromelon TM chromatography data system with P580 and UVD100 detectors (Thermo Fisher Scientific Inc., Waltham, MA, USA). Chromatographic separation was performed on a Discovery C18 (250 × 4.6 mm, 5-µm particle size). Column temperature was 25 °C; flow rate was 1.0 mL/min; injected volume was 10 µL.
3
Quantification of Camu-Camu Bioactive Compounds
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The method to identify active components of camu-camu fruit extract was described in a previous study [19 (link)]. In brief, HPLC was operated on a Dionex Chromelon TM chromatography data system with P580 and UVD100 detectors (Thermo Fisher Scientific Inc., Waltham, MA, USA). Chromatographic separation was performed on a SUPELCO, Discovery C18 column (250 × 4.6 mm, 5 µm particle size); flow rate, 1 mL/min; injection volume, 10 µL, column temperature, 30 °C. An ascorbic acid standard was prepared at concentrations between 1 μg/mL and 500 μg/mL. The chromatographic condition for ascorbic acid was set as follows: 2.5% ethanol in 25 mmol/L sodium dihydrogenphosphate (v/v); detection wavelength, 256 nm. The cyanidin-3-glycoside standard was diluted in a range of 0.1 to 50 μg/mL. The mobile phase was composed of (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile. The gradient was linearly increased from 5% B to 95% B over 40 min. The absorbance wavelength of the cyanidin-3-glycoside was detected at 520 nm.
4
HPLC Quantification of Chlorogenic Acid and Epicatechin
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High-performance liquid chromatography (HPLC) was performed on a Dionex Chromelon TM chromatography data system with P580 and UVD100 detectors (Thermo Fisher Scientific Inc., Waltham, MA, USA). Analyzed sample (CLE) was prepared in 50% methanol at 5mg/mL. Ranges of diluted concentrations (3.125–100 µg/mL) of standard chlorogenic acid and (−)-epicatechin were dissolved in 50% methanol. Chromatographic separation was performed on a Waters 120 ODS-AP (250 × 4.6 mm2, 5-µm particle size). Elution was performed with a water/acetonitrile (3:1) gradient containing 1% formic acid. The gradient was linearly increased from 5% to 90% acetonitrile over 35 min. The injection volume was 10 µL and the flow rate was 1 mL/min. Analysis of chlorogenic acid and (−)-epicatechin in CLE was validated by using the external standard method, according to the International Conference on Harmonization [56 ] (see Supplementary Materials ). Linear correlations (R2) of the calibration curves for each compound were 0.999. The limit of detections (LOD) and limit of quantifications (LOQ) were less than 0.028 µg/g and 0.074 µg/g, respectively (see Supplementary Materials ). Thus, established HPLC method was suitable for the quantitative analysis.
5
HPLC Analysis of Plant Extract Compounds
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The plant extract was prepared in 50% methanol at a concentration of 2 mg/ml. Serial dilutions (2.5, 25, 125, 250, 500, and 1000 µg/mL) of standard compounds (tannic acid, chlorogenic acid, and apigenin) were prepared in methanol. High-performance liquid chromatography (HPLC) was performed on a Dionex Chromelon TM chromatography data system with P580 and UVD100 detectors (Thermo Fisher Scientific Inc., Waltham, MA, USA). Chromatographic separation was performed on a Discovery C18 (250 × 4.6 mm, 5-µm particle size). Column temperature was 25 °C; flow rate was 1.0 mL/min; injected volume was 10 µL. The condition for chromatographic separation was described in Supplementary Materials Table S1 . The chemical content was quantified by determining the area of the peak in the HPLC analysis, following the formula below:
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