RP-HPLC method was used to determinate phenolic compounds by an Agilent 1260 series HPLC instrument (Agilent Technologies, Palo Alto, CA, USA) equipped with a quaternary pump (G1311B), degasser, column thermostat (G1316A), auto-sampler (G1329B) and diode array detector (G1315 B, DAD). Chromatographic separation was carried out according to Molinu et al. [20 (link)]. The column was a Zorbax Eclipse plus C18 (250 × 4.6 mm, 5 µm; Agilent). Column temperature was set to 30 °C and the flow rate was 0.8 mL min−1. The injection volume was 10 μL and the detection wavelengths were set to 280, 330 and 350 nm. Data were processed using the Agilent OpenLAB CDS ChemStation edition 2012. Peak assignment of phenolic compounds was carried out comparing their retention times and UV-vis spectra with those of analytically pure standard (Table 7 ), and as well as by adding the standard solution to the sample.
Agilent 1260 series hplc instrument
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1260 series HPLC instrument is a high-performance liquid chromatography system designed for analytical and preparative separations. It features a modular design that allows for the integration of various components, including pumps, autosamplers, column compartments, and detectors, to create a customized solution for a wide range of applications.
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Lab products found in correlation
Openlab cds chemstation edition 2012, by Agilent Technologies (2 mentions)
Zorbax eclipse plus c18, by Agilent Technologies (2 mentions)
Hss t3, by Waters Corporation (1 mentions)
Agilent openlab cds chemstation software, by Agilent Technologies (1 mentions)
Dad detector, by Agilent Technologies (1 mentions)
Zetasizer nano zs90, by Malvern Panalytical (1 mentions)
Ht7700, by Hitachi (1 mentions)
Su8010, by Hitachi (1 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
10 protocols using agilent 1260 series hplc instrument
1
Quantifying Phenolics by RP-HPLC
2
HPLC Analysis of Drug-Loaded Nano-Microcapsules
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The LC and Chl drug loading contents were determined using an Agilent 1260 series HPLC instrument fitted with a Zorbax Carbon hydrate analysis column (150 mm long, 4.6 mm i.d., 5 µm particle size; Agilent Technologies). The HPLC settings are shown in Table 1 . A specified amount of the freeze-dried nano-microcapsules was dissolved in tetrahydrofuran; then, the LC and Chl concentrations were determined by HPLC. The encapsulation efficiency was determined by determining the LC and Chl concentrations in the nano-microcapsules and supernatant after centrifugation using the same method. The equations used to calculate the drug loading content and encapsulation efficiency are shown below.
3
HPLC Analysis of Marker Compounds
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An Agilent 1260 series HPLC instrument (Agilent, USA) equipped with a DAD detector (Agilent, USA) was used for chromatographic analysis of quantitative determination of the three marker compounds. The system control and data analysis were processed using Agilent OpenLAB CDS ChemStation software (Agilent, USA). The chromatographic separation was performed on a Waters HSS T3 (4.6 × 250 mm, 5 μm, Waters, USA) column at 40°C: (A) 0.3% phosphorous acid in water and (B) acetonitrile as mobile phases. The sample was injected (5 μL injection volume) onto the column and eluted at a flow rate of 1 mL/min according to the following gradients: initial 16% B; 0–40 min/16–18% B; 40–43 min/18–80% B; and 43–48 min/80–80% B. Ultraviolet detection was set to 330 nm.
4
HPLC Analysis of Phenolic Compounds
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Analyses of individual phenolic compounds were carried out using an Agilent 1260 series HPLC instrument (Agilent Technologies, Palo Alto, CA, USA) equipped with a quaternary pump, degasser, column thermostat, auto-sampler and diode array detector. Chromatographic separation was carried out according to Molinu et al. [52 (link)]. The column was a Zorbax Eclipse plus C18 (250 × 4.6 mm, 5 µm; Agilent). Column temperature was set to 30 °C and the flow rate was 0.8 mL min−1. The injection volume was 10 μL and the detection wavelengths were set to 280, 330 and 350 nm. Data were processed using the Agilent OpenLAB CDS ChemStation edition 2012. The concentration of six phenolic compounds (neoclorogenic acid, clorogenic acid, criptoclorogenic acid, verbascoside, diosmin, luteolin) were carried out comparing their UV-vis spectra and retention times with those of analytically pure standard.
5
Nano-microcapsule Characterization and Analysis
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The nano-microcapsules were dispersed in deionized water and diluted to a concentration of 0.5%. The morphological features of the nano-microcapsules were investigated by scanning electron microscopy (SEM), using an SU8010 instrument (Hitachi, Tokyo, Japan) with an acceleration voltage of 5 kV, and by transmission electron microscopy (TEM), using an HT7700 instrument (Hitachi, Tokyo, Japan) with a voltage of 80 kV. The nano-microcapsule particle sizes and polydispersity index (PDI) were determined using a dynamic laser scattering (DLS) instrument (Zetasizer Nano ZS90, Malvern Panalytical, Malvern, UK). Each sample was analyzed in triplicate to ensure that the data were reliable. The nano-microcapsules were analyzed by Fourier-transform infrared spectroscopy using the KBr method. The LC and Chl concentrations were determined by high-performance liquid chromatography (HPLC) using an Agilent 1260 series HPLC instrument (Agilent Technologies, Santa Clara, CA, USA). Environmental SEM images of the nano-microcapsules were acquired using an FEI Quanta FEG 250 SEM instrument (Thermo Fisher Scientific, Waltham, MA, USA).
6
Stability Analysis of SSN Granules
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The SSN granule was manufactured by Deyuantang Industry Co. Ltd, Shanxi, China. To verify the stability of SSN granule’s composition, ten different batches of SSN granules were tested using HPLC method. A 100 mg SSN granule was added into 2 ml methanol ultrasonic extraction and incubated for 30 min, then filtered through a 0.45-μm nylon membrane filter for HPLC analysis. Chromatographic analysis was performed on an Agilent 1260 series HPLC instrument (Agilent, Inc., MA, U.S.A.) coupled to a binary pump (G1311C), an autosampler (G1329B), a thermostatted column compartment (G1316A), and a UV detector (G4212B-DAD). The sample was separated on a SunFire™ C18 (250 mm × 4.6 mm, 5 μm) with the column temperature maintained at 25°C. The analytical separation was run using a gradient elution composed of solvent A (acetonitrile) and solvent B (0.1% formic acid). A linear gradient elusion was used as follows: 0–1 min, 5–23% A; 1–18 min, 23–25% A; 18–19 min, 25–30% A; 19–31 min, 30–75% A; 31–60 min, 75–85% A; 60–90 min, 85–100% A. The flow rate was 1.0 ml/min and the UV spectra were set at 254 nm. The injection volume was 5 μl.
7
HPLC Analysis of GRR and GRR-CDs
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High-performance liquid chromatography (HPLC) was used to evaluate the individual components before and after pyrolysis and dialysis to compare the difference between GRR and GRR-CDs. The fingerprint analysis was conducted using a ZORBAX-C18 column (250 × 4.6 × 5 mm, Orochem Technologies, Illinois, USA) based on an Agilent series 1260 HPLC instrument (Agilent Technologies, Waldbronn, Germany) equipped with a quaternary pump, a diode-array detector, a degasser, an autosampler, as well as a column compartment. The methanol extract of GRR and GRR-CDs was prepared and then processed under the same detection conditions. The mobile phase A (acetonitrile) and B (4% phosphoric acid) were filtered through 0.22 µm cellulose acetate membrane filters (JinTeng, Tianjin, China) prior to use. The gradient program is elucidated in the following: 0–15 min, 10–20% A and 90–80% B; 15–30 min, 20% A and 80% B; 30–40 min, 20–30% A and 80–70% B; 40–50 min, 30–10% A and 70–90% B. Furthermore, the column was held at 35 °C; the flow rate was set to 1.0 mL/min; and the detection wavelength was 254 nm.
8
Quantitative HPLC Analysis of Compounds
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HPLC analysis was performed on an Agilent series 1260 HPLC instrument (Agilent, Germany) consisting of a quaternary pump, a Diode Array Detector (DAD), an autosampler and a column compartment. Chromatographic separations were performed on an Agilent Zorbax SB C18 column (250 mm × 4.6 mm, 5 μm, USA). The mobile phase consisted of acetonitrile (B) and 0.2% (v/v) formic acid in water (A) with a gradient of 10–50% B at 0–65min, 50–10% B at 65–70 min. The analysis was performed at 30 °C with a flow-rate of 1 mL/min. The detection wavelength was 280 nm. An aliquot of 20 μL was injected for HPLC. The data were processed with Agilent Technologies ChemStation Revision B.04.03 software.
9
Microbial Functional Capacity and Vitamin K
10
HPLC-ELSD Gradient Separation for Compound Analysis
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The analysis was performed on an Agilent series 1260 HPLC instrument coupled with an Agilent 1290 ELSD (Agilent Technologies, Santa Clara, CA, USA). The system was equipped with Diamonsil TM C18 column (250 mm × 4.6 mm, 5 μm) with the flow rate of 1.0 mL/min at 35 °C. The mobile phase was 0.1% acetic acid water (A) and methanol (B) with a gradient elution of 10–25% B at 0–20 min, 25–45% B at 20–50 min, 45–55% B at 50–55 min, 55–65% B at 55–70 min, 65–70% B at 70–90 min, 70–85% B at 90–115 min, 85–95% B at 115–125 min, 95–10% B at 125–130 min. The drift tube temperature of ELSD was 70 °C and nitrogen cumulative flow rate was 1.6 L/min.
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