The conversion of FF, NF, MAL, and PA was quantified relative to the unheated samples (t = 0 min) by HPLC-DAD. Samples were diluted (1:20) and analyzed with the following setup: degasser, Shimadzu DGU-20As; pump, Shimadzu LC-20AD; autosampler, Shimadzu SIL-10AF; column oven, Shimadzu CTO-20A; column, Nucleosil® 120-5 C18 (Macherey-Nagel GmbH & Co. KG, Düren, Germany); detector Shimadzu SPD-M20A; software, Shimadzu LabSolutions Version 5.90. The following settings were used: column temperature, 45 °C; flow rate, 1.0 mL/min; eluent A, 0.075% acetic acid in water (v/v); eluent B, methanol; eluent gradient, 0 min, 5% B; 10 min, 20% B; 15 min, 90% B; 20 min, 90% B; 21 min, 5% B; wavelength for quantitation, 285 nm.
Dgu 20a
Manufactured by Shimadzu
Sourced in Japan, Germany
The DGU-20A is a degassing unit designed for use with high-performance liquid chromatography (HPLC) systems. Its core function is to remove dissolved gases from the mobile phase, which helps to improve the stability and performance of the HPLC system.
Automatically generated - may contain errors
Lab products found in correlation
Lc 20ad, by Shimadzu (9 mentions)
Spd m20a, by Shimadzu (8 mentions)
Cto 20a, by Shimadzu (8 mentions)
Cbm 20a, by Shimadzu (7 mentions)
Sil 20ac, by Shimadzu (6 mentions)
Sil 30ac, by Shimadzu (5 mentions)
Lc 30ad, by Shimadzu (5 mentions)
Spd 20a, by Shimadzu (5 mentions)
Hplc system, by Shimadzu (4 mentions)
Cto 20ac, by Shimadzu (4 mentions)
Lc 20ab, by Shimadzu (4 mentions)
Cto 10a, by Shimadzu (3 mentions)
Lc 20at, by Shimadzu (3 mentions)
Sil 20a ht, by Shimadzu (3 mentions)
Labsolutions version 5, by Shimadzu (2 mentions)
Sil 10af, by Shimadzu (2 mentions)
Nucleosil 120 5 c18, by Macherey-Nagel (2 mentions)
Cto 20a, by Macherey-Nagel (2 mentions)
Gemini c18 column, by Phenomenex (2 mentions)
Sil 20a, by Shimadzu (2 mentions)
37 protocols using dgu 20a
1
HPLC-DAD Quantification of Phytochemical Conversions
2
Quantitative Ginkgo Biloba Analysis
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The Prominence LC-20 series (Shimadzu) chromatograph with a diode-matrix detector SPD20 M, autosampler SIL-20AS, pumps LC-20AD, degasser DGU-20 As, column thermostat CTO-20A, detector with diode matrix SPD-M20A, fluorometric detector, and CBM-20A system controller was used to analyze the component content of aqueous–alcoholic extracts of Gingko biloba. In this study, LC (Shimadzu, Corporation) software, version 5.23 SP1, was used. A Phenomenex Gemini C18 column (110 Å, 5 µm, 4.6 mm×250 mm) was used as a stationary phase, with a mobile phase consisting of deionized water–acetonitrile with the addition of formic acid. The following mixtures were used: water–acetonitrile 95:5 + 0.1% formic acid (eluent A) and acetonitrile–water (95:5) + 0.1% formic acid (eluent B). The flow rate was 0.8 mL/min, at a temperature of 40 °C. The separation was carried out in the gradient elution mode according to [53 (link)]. Injection volumes were 20 µL, in five repetitions. Relative retention times were determined using flavonoid and terpene lactone standards (Acros, J and K, Aldrich-Sigma). The quantitative determination of the isolated components was performed under the HPLC method conditions. A diode-matrix detector with a wavelength range of 190–750 nm was used for this research.
3
Serum Retinol Levels in Maternal and Offspring Rats
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The maternal rats at preconception and five to six pups from each group at postnatal 24 hours, 4 weeks, 8 weeks, 12 weeks, 18 weeks, 22 weeks and 26 weeks were randomly selected to detect serum retinol levels. The pups younger than 8 weeks old were decapitated to collect the blood; and the blood from the pups at the age of 8 weeks and older were collected via the caudal vein. All pups were decapitated after the last behavioral tests to detect serum retinol levels and confirm the success of the animal models. Serum retinol was measured using high performance liquid chromatography (HPLC) as previously described [31 (link)]. Briefly, 200 µl of serum was deproteinized with an equal volume of dehydrated alcohol. The retinol was extracted from the serum by adding 1,000 µl of hexane and evaporated with nitrogen gas. The retinol residue was dissolved in 100 µl of the mobile phase mixture (methanol:water=97:3). Then, 20 µl of the dissolved liquid was tested in the HPLC apparatus (DGU-20As, Shimadzu Corporation, Kyoto, Japan) on a silica column with a 315-nm ultraviolet photodiode array detector. All procedures were performed in a dark room to protect the samples from light at the Children’s Nutrition Research Center in the Children’s Hospital of Chongqing Medical University.
4
Analytical HPLC characterization of peptides
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Lyophilized peptides and XCL1 were dissolved at 1 mg·mL-1 in ACN:ddH2O (1:1 v/v) supplemented with 0.01% TFA, and analytical HPLC was carried out on Shimadzu instrument composed of a CBM-20A communication Bus module, DGU-20AS degasser, 2 LC20AD pumps, SIL-20AC autosampler, SPD-M20A diode array detector and CTO-20AC column oven. For XCL1(CC3) and S7Abu peptides, a linear gradient from 5% to 95% of ACN (+0.036% TFA) into ddH2O (+0.045% TFA) were run at flow rate of 1 mL·min-1 over 15 min. For Y7A, R7A and F7M peptides, a linear gradient from 5% to 95% of ACN (+0.01% TFA) into ddH2O (+0.01% TFA) were run at flow rate of 1 mL·min-1 over 37 min. XSelect Peptide CSHTM C18, 130 Å, 3.5 µm, 4.6 mm × 100 mm (Waters) column was used for the analysis. Chromatographic peak area was determined at λ=220 nm.
5
Serum Retinol Quantification by HPLC
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Serum retinol concentrations were measured using high-performance liquid chromatography (HPLC) according to our previously described methods [20] (link), with slight modifications. Briefly, 200 µl of serum was deproteinized with an equal volume of dehydrated alcohol. A total of 1,000 µl of hexane was used to extract the retinol from the serum; the hexane was then evaporated using nitrogen gas. The retinol residue was dissolved in 100 µl of the mobile phase mixture (methanol: water = 97∶3). Finally, the prepared sample was measured using an HPLC apparatus (DGU-20As, Shimadzu Corporation, Kyoto, Japan) on a C18 analytical column with a 315-nm ultraviolet photodiode array detector.
6
Measuring Serum Retinol by HPLC
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Serum retinol concentrations were measured using high-performance liquid chromatography (HPLC) according to our previously described methods (14 (link)), with slight modifications. Briefly, 200 mL of serum was deproteinized with dehydrated alcohol, and then retinol was extracted with hexane and evaporated with nitrogen gas. The residue of retinol was dissolved in 100 μL of the mobile phase mixture (methanol: water = 97:3). Finally, the prepared sample was measured using an HPLC apparatus (DGU-20As, Shimadzu Corporation, Japan) equipped with a C18 analytical column and a 315 nm ultraviolet photodiode array detector.
7
Liquid Chromatography-Mass Spectrometry Quantitation of Vitamin K Derivatives
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LC-MS/MS was performed using an LCMS-8050 Liquid Chromatograph Mass Spectrometer (Shimadzu, Kyoto, Japan) and Shimadzu HPLC System (system controller (CBM-20A), pump (LD-20AD), degasser (DGU-20As), UV detector (SPD-20A), and auto injector (SIL-20AC HT)). Separations were performed on a CAPCELL PAK C18 UG120 (3 μm, 2.0 mm × 100 mm, Shiseido Co., Ltd., Tokyo, Japan) using a mobile phase of 10 mmol/L ammonium acetate and 0.1% acetic acid in methanol and water (97:3) at a flow rate of 0.4 mL/min. Column temperature was maintained at 40 °C. The mass spectrometer was equipped with an electrospray ionization and was run in positive ion mode. Identification and quantitation were based on MS/MS-multiple reaction monitoring mode using transition ions as follows: m/z 445 → 187 for the [M+H]+ MK-4 adduct, m/z 445 → 187 for the [M + H]+ chromenol adduct, 619 → 58 for the [M + H]+ MKH-DMG adduct, m/z 532 → 58 for the [M + H]+ MKH-mono-DMG adduct, m/z 664 → 187 for the [M + H]+ MKH-SUC adduct, m/z 564 → 187 for the [M + H]+ MKH-mono-SUC adduct, and m/z 461 → 81 for the [M + H]+ MKO adduct. Retention times were: MK-4, 3.3 min; MK-4 chromenol, 2.0 min; MKH-DMG, 1.7 min; MKH-mono-DMG, 1.6 min; MKH-SUC, 1.1 min; MKH-mono-SUC, 1.2 min; and MKO, 2.5 min.
8
Serum Retinol Determination by HPLC
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The serum retinol levels in the collected mouse blood were determined using HPLC. VA standard curve preparation and testing methods were modified slightly following methods described previously (Li et al., 2017 (link)), and VA standard compound was purchased from Sigma (R7632, United States). Briefly, 200 μL of serum was deproteinized with the same volume of anhydrous ethanol. Then, 1000 μL of hexane was used to extract the retinol from the serum, and the hexane was evaporated using nitrogen gas. The retinol residue was dissolved in 100 μL of the mobile phase mixture (methanol:water = 97:3). Finally, the prepared sample was measured using an HPLC apparatus (DGU-20As, Shimadzu Corporation, Japan). The retinoids were separated by chromatography on an analytical column (Hypersil phenyl 120 A 5 mm, 250 mm × 4.6 mm, Phenomenex, United States) via gradient elution of the mobile phase in a liquid chromatograph equipped with a 315-nm ultraviolet photodiode array detector.
9
UHPLC Quantification of Polyphenol Compounds
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Phenolic compounds were extracted following the International Oleic Council method [18 ] and hydrolyzed according to the method proposed by Rovellini et al. [19 ]. The hydrolyzed sample was then analyzed by UHPLC using an Agilent Poroshell 120 EC-C18 reversed-phase column (2.7 µm particle size, 4.6 × 150 mm) on a Shimadzu Nexera UHPLC System (Shimadzu Nexera, Kyoto, Japan) equipped with dual pump LC-30AD, on-line degasser DGU-20AS, column oven CTO-30A, autosampler SIL-30AC and diode array detector (SPD-M20A). Gradient separation was created from solvent A (water with 2% of acetic acid) and solvent B (acetonitrile) as follows: starting from 95% A; 0.01–12 min linear gradient from 5% to 70% B; 12–13 min linear gradient from 70% to 90% B; isocratic condition kept up to 17 min; 17 min back to initial condition at 5% B; isocratic step kept up to 22 min for column re-conditioning. The mobile phase flow rate was 450 μL min−1. The column temperature was 30 °C. Injected volumes for each sample was 5 μL. The detector was set at 280 nm. Polyphenols quantification was obtained using calibration curves obtained by injection on the column of different amounts of both tyrosol and hydroxytyrosol (10–600 ng) with R2 values higher than 0.999, in all cases.
10
Quantification of Methylglyoxal via HPLC-DAD
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MGO was analyzed as its corresponding quinoxaline derivative and quantified relative to the unheated samples (t = 0 min) by HPLC-DAD [41 (link)]. For derivatization, aliquots of the model systems were incubated with ortho-phenylendiamine (OPD): 0.1 mL of OPD solution (50 mmol/L in water/methanol 1:1, v/v) was added to the sample (0.1 mL). Subsequently, samples were stored in darkness at room temperature for 24 h. After derivatization, samples were diluted (1:50) and analyzed by HPLC-DAD. The following setup was used: degasser, Shimadzu DGU-20As; pump, Shimadzu LC-20AD; autosampler, Shimadzu SIL-10AF; column oven, Shimadzu CTO-20A; column, Nucleosil® 120-5 C18 (Macherey-Nagel GmbH & Co. KG, Düren, Germany); detector Shimadzu SPD-M20A; software, Shimadzu LabSolutions Version 5.90. The following settings were used: column temperature, 35 °C; flow rate, 0.5 mL/min; eluent A, 0.075% acetic acid in water (v/v); eluent B, methanol; eluent gradient, 0 min, 40% B; 10 min, 40% B; 15 min, 60% B; 25 min, 60% B; 26 min, 90% B; 34 min, 90% B; 35 min, 40% B; wavelength for quantitation, 318 nm.
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