Tocopherols in the seed oils were quantified using the standard method of the agricultural industry of China (NY) (NY/T 1598-2008) with slight modifications. Tocopherols were analyzed by a Shimadzu (Kyoto, Japan) LC2010HT liquid chromatographic system comprising two LC-10ADvp pumps, an SIL-10ADvp autosampler, an SPD-10A(V)vp ultraviolet-visible detector, a CTO-10a(C)vp column oven, and an LC solution work station. HPLC analysis was performed on a Global Chromatography GS-120-5 C18 column (250 × 4.6 mm, 5 μm, Unimicrotech). The mobile phase was methanol-ultrapure water, 98 : 2 (v/v), at a flow rate of 1 mL/min. The column temperature was maintained at 30°C. The detection wavelength was 294 nm. The volume of sample injected was 10 μL. Quantification was performed based on calibration using standard tocopherols.
Sil 10advp autosampler
Manufactured by Shimadzu
Sourced in Japan
The SIL-10ADvp autosampler is a component of Shimadzu's high-performance liquid chromatography (HPLC) systems. Its core function is to automatically inject samples into the HPLC system for analysis.
Automatically generated - may contain errors
Lab products found in correlation
Lc 10advp pump, by Shimadzu (3 mentions)
Security guard column, by Phenomenex (2 mentions)
Lc 10at vp, by Shimadzu (2 mentions)
Lc 2010ht, by Shimadzu (1 mentions)
Lc 10, by Shimadzu (1 mentions)
Hplc elsd system, by Shimadzu (1 mentions)
Dgu 14a, by Shimadzu (1 mentions)
Spd m10avp dad detector, by Shimadzu (1 mentions)
Scl 10a system controller, by Shimadzu (1 mentions)
Cto 10avp column oven, by Shimadzu (1 mentions)
Eclipse xdb c18, by Agilent Technologies (1 mentions)
Dgu 14a degasser, by Shimadzu (1 mentions)
Rf 10axl fluorescence detector, by Shimadzu (1 mentions)
Autosampler series 3000, by Agilent Technologies (1 mentions)
Spd m10a vp diode array detector, by Shimadzu (1 mentions)
High performance liquid chromatography (hplc), by Shimadzu (1 mentions)
Luna 5 m c18 column, by Phenomenex (1 mentions)
Cto 10, by Shimadzu (1 mentions)
Spd m20a, by Shimadzu (1 mentions)
Lc 20ad pump, by Shimadzu (1 mentions)
12 protocols using sil 10advp autosampler
1
Quantification of Seed Oil Tocopherols
2
RP-HPLC Analysis of Glutathione Metabolites
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The levels of the reduced (GSH) and oxidized (GSSG) glutathione, cysteine and cystine in the incubation mixtures were determined using the RP-HPLC method of Dominic et al. [27 (link)] with modifications [28 (link)]. The samples were separated on a 4.6 mm × 250 mm Luna C18 (5 µm) column with a Phenomenex Security Guard column filled with the same packing material. The chromatographic system consisted of LC-10 Atvp Shimadzu Corp. pumps, four channel degassers, column oven, a Shimadzu SIL-10 Advp autosampler and a Shimadzu Corp. SIL-10 SPD-M10Avp-diode array detector; Lab Solution LC software was used to control system operation and facilitate data collection. The standard curves were generated in the supernatant obtained from cellular homogenates in the range from 13 to 75 nM of each compound per ml. All the standard curves generated for the analyte were linear in the investigated concertation range.
3
HPLC-ELSD for Disaccharide Separation
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To enable separation of individual disaccharides, the honey samples were chromatographed on a Shimadzu HPLC-ELSD system consisting of a Shimadzu Class VP Pump LC-10AD VP/Valve FCL-10AL VP/Degasser DGU-14A with a Class VP Software/ SCL-10A VP controller, SIL-10AD VP autosampler and a CTO-10A VP column oven operated at 40 °C. Separations were performed on a Phenomenex Luna 5 µm NH2 100 Å 250 × 4.6 mm column with an isocratic mobile phase comprised of 85% acetonitrile and 15% RO water at a flow rate of 2.5 mL/min. Eluted sugars were monitored with a Shimadzu ELSD- LT (Low Temperature) detector operated at 350 kPa and 45 °C. The elution flow was diverted to a collection tube when the target peak emerged and 5 mL (2 min) fractions collected before reconnection of the column eluant to the ELSD detector (to see the tail end of the peak). Collected fractions were analysed ‘as is’ by UPLC-MS/MS (as above), and a second portion freeze-dried and dissolved in D2O for NMR analysis.
4
Serotonin Quantification in Rat Brain Regions
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Rats were sacrificed by decapitation and their brains were rapidly removed. Various brain areas, including cortex, striatum, and hippocampus were rapidly dissected on an icy cold plate, then weighed and stored at -80°C until homogenization with ultrasonication in 0.2 mL of 7N perchloric acid (Sigma Chemical Industries, Ltd., Saint Louis, MO, USA). Homogenates were centrifuged at 12,000×g for 30 min at 4°C.
The concentration of serotonin (5-HT) was determined by high performance liquid chromatography (HPLC) equipped with electrochemical detector (ECD, LC-4C, BAS, West Lafayette, IN, USA) (10 nA, filter 2.0 Hz, AppE cell 0.750 V), autosampler (Shimadzu SIL-10ADvp autosampler, Shimadzu, Japan). The supernatant was filtered through a 0.22 um filter, and was analyzed by HPLC equipped with an AlltimaTM (Grace Davison Discovery Scienes, Ltd., Deerfield, IL, USA) reversed-phase C18 column (4.6×150 mm, 5 μm). Injection volume was 20 uL. The mobile phase contains 100 mM NaH2PO4.H2O, 0.74 mM sodium octanesulfonate (SOS), 0.02 mM EDTA and 20% methanol adjusted the pH to 3.0 by H3PO4 and with a flow rate of 1.0 mL/min. The calibration curve was obtained based on 6 levels (20, 40, 60, 80, 100, and 200 ppm) of 5-HT standard (Sigma Chemical Industries, Ltd.). The coefficient of determination (r2) was greater than 0.995.
The concentration of serotonin (5-HT) was determined by high performance liquid chromatography (HPLC) equipped with electrochemical detector (ECD, LC-4C, BAS, West Lafayette, IN, USA) (10 nA, filter 2.0 Hz, AppE cell 0.750 V), autosampler (Shimadzu SIL-10ADvp autosampler, Shimadzu, Japan). The supernatant was filtered through a 0.22 um filter, and was analyzed by HPLC equipped with an AlltimaTM (Grace Davison Discovery Scienes, Ltd., Deerfield, IL, USA) reversed-phase C18 column (4.6×150 mm, 5 μm). Injection volume was 20 uL. The mobile phase contains 100 mM NaH2PO4.H2O, 0.74 mM sodium octanesulfonate (SOS), 0.02 mM EDTA and 20% methanol adjusted the pH to 3.0 by H3PO4 and with a flow rate of 1.0 mL/min. The calibration curve was obtained based on 6 levels (20, 40, 60, 80, 100, and 200 ppm) of 5-HT standard (Sigma Chemical Industries, Ltd.). The coefficient of determination (r2) was greater than 0.995.
5
Serotonin and Metabolite Analysis in Rat Brain
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Rats were sacrificed by decapitation and their brains were rapidly removed. Various brain areas, including prefrontal cortex, hippocampus, and amygdala were rapidly dissected on an icy cold plate, then weighed and stored at -80°C until homogenization with ultrasonication in 0.2 mL of 7N perchloric acid (Sigma Chemical Industries, Ltd., Saint Louis, MO, USA). Homogenates were centrifuged at 12,000×g for 30 min at 4°C. The concentration of serotonin (5-HT) and its metabolite 5-HIAA was determined by high performance liquid chromatography (HPLC) equipped with electrochemical detector (ECD, LC-4C, BAS, West Lafayette, IN, USA) (10 nA, filter 2.0 Hz, AppE cell 0.750 V), autosampler (Shimadzu SIL-10ADvp autosampler, Shimadzu, Japan). The supernatant was filtered through a 0.22 um filter, and was analyzed by HPLC equipped with an AlltimaTM (Grace Davison Discovery Scienes, Ltd., Deerfield, IL, USA) reversed phase C18 column (4.6×150 mm, 5 μm). Injection volume was 20 uL. The mobile phase contains 100 mM NaH2PO4.H2O, 0.74 mM sodium octanesulfonate (SOS), 0.02 mM EDTA and 20% methanol adjusted the pH to 3.0 by H3PO4 and with a flow rate of 1.0 mL/min. The calibration curve was obtained based on 6 levels (20, 40, 60, 80, 100, and 200 ppm) of standard of 5-HT and 5-HIAA (Sigma Chemical Industries, Ltd.). The coefficient of determination (r2) was greater than 0.995.
6
Quantification of Sulfur-Containing Compounds
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The RP-HPLC method of Dominick et al. [49 (link)] with the modifications described by Bronowicka-Adamska et al. [50 (link)] was used to determine the level of low molecular weight sulfur-containing compounds, such as reduced (GSH) and oxidized (GSSG) glutathione, cysteine and cystine (the chromatographic system consisted of LC-10 Atvp Shimadzu Corp. (Kyoto, Japan) pumps, four channel degassers, column oven and a Shimadzu Corp. SIL-10 Advp autosampler; the chromatographic peaks were measured by a Shimadzu Corporation SPD-M10Avp-diode array detector; LabSolution (Warsaw, Poland) LC software was used to control system operation and facilitate data collection). The standard curves were generated in the supernatant obtained from cellular homogenates in the range from 13 to 75 nM of each compound per mL. All the standard curves generated for the analyte were linear in the investigated concentration range.
7
Phenolic Profiling of Olive Pomace Extract
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Phenolic profiles of OPE were determined by HPLC coupled to a diode array (HPLC–DAD). Standard calibration curves were prepared by using gallic, protocatechuic acid, catechin, p-hydroxybenzoic, syringic, elagic, m-coumaric, o-coumaric, myricetin, quercetin, kaempferol, hydroxytyrosol, tyrosol, and luteolin. The HPLC analysis was performed using a modified method [5 (link)]. The samples and stock solutions were filtered through a 0.45 µm membrane filter and analyzed in a Shimadzu HPLC system (LC-10AD vp pump, SPDM10A vp DAD detector, SIL-10AD vp autosampler, CTO-10AVP column oven, DGU-14A degasser, and SCL-10A system controller; Shimadzu Corp., Kyoto, Japan). Separations were performed at 30 °C on Agilent Eclipse XDB-C18 reversed-phase column (250 mm × 4.6 mm length, 5 μm particle size). The mobile phase contained solvent A (3% (v/v) acetic acid) and solvent B (methanol). A gradient elution was carried out as shown: 28% B (0–20 min), 28–30% B (21–50 min), 31–50% B (51–70 min), and 50–100% B (70–81 min) and at 90 min was returned to initial conditions. The flow rate was 0.8 mL/min. Chromatograms were recorded at 278 nm. Identification and quantitative analysis were made based on the retention times and external standard curves. The amounts of polyphenols were stated in μg/g of dried olive pomace extract.
8
Contaminant Accumulation and Enzymatic Response
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Water samples were taken at days 0, 14, and 21 after addition of the contaminants and were analyzed for their concentrations of metals and organic contaminants. The soluble metals were determined by inductively coupled plasma mass spectrometry (7500cx coupled to Autosampler Series 3000, both from Agilent Technologies). BPA, CIP, and SMX concentrations were measured by high-performance liquid chromatography (HPLC) (Shimadzu Corp., Kyoto, Japan), equipped with LC-10 ADVP solvent delivery module, SPD-M10 AVP Diode Array Detector, RF-10AXL Fluorescence Detector, and SIL-10 ADVP autosampler. Separation of BPA was accomplished on a Nucleosil 100–5 C-18 column and separation of CIP and SMX was performed on an Alltech PrevailTM Organic Acid 5u as previously described by Christofilopoulos et al. (2016) (link).
At the end of the experiment, the plants were harvested and washed first with water and then with distilled water. The fresh weights of roots and leaves were determined, plant parts were cut in small pieces and 0.4 g of each plant compartment was sampled for enzymatic analysis while 0.3 g of roots was taken for DNA extraction. The plant samples for further analysis were immediately snap-frozen in liquid nitrogen and stored at -80°C; the remaining material was dried at 45°C and weighed.
At the end of the experiment, the plants were harvested and washed first with water and then with distilled water. The fresh weights of roots and leaves were determined, plant parts were cut in small pieces and 0.4 g of each plant compartment was sampled for enzymatic analysis while 0.3 g of roots was taken for DNA extraction. The plant samples for further analysis were immediately snap-frozen in liquid nitrogen and stored at -80°C; the remaining material was dried at 45°C and weighed.
9
Quantitative Analysis of Redox Biomarkers
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In order to determine the levels of the reduced (GSH) and oxidized (GSSG) glutathione, cysteine, and cystine, the RP-HPLC method of Dominic and others [53 (link)], with modifications described by Bronowicka-Adamska and others [54 (link)], was used. The samples were separated on a 4.6 mm × 250 mm Luna C18 (5 µm) column (Phenomenex, Warsaw, Poland) with a Phenomenex Security Guard column filled with the same packing material. The chromatographic system consisted of LC-10 Atvp Shimadzu Corp. pumps, four channel degassers, column oven, a Shimadzu SIL-10 Advp autosampler, and a Shimadzu Corp. SIL-10 SPD-M10Avp-diode array detector (Shim-Pol, A.M. Borzymowski, Warsaw, Poland); LabSolutions LC software was used to control the system operation and facilitate data collection. The standard curves were generated in the supernatant, which was obtained from cellular homogenates in the range from 13 to 75 nM of each compound per ml. All the standard curves generated for the analyte were linear in the investigated concertation range.
10
Quantification of Phenolic Compounds
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The procedure for the phenolic contents has been described by Capanio et al. (1999) . Highperformance liquid chromatography was used. Detection and quantification was carried out with a SLC-10Avp system controller (Shimadzu, Japan) SIL-10 AD vp Autosampler, LC-10 AD vp pump, DGU-14a degasser, CTO-10 A vp column heater and diode array detector set at 278 nm. AnAgilent Eclipse XDB C-18 column (250x4.6mm,5mikrometre) was used. The flow rate was 0.8 mL/min, injection volume was 10 microlitre and the column temperature was set at 30°C. Methanol and 3% acetic acid were used for mobile phases. The data were integrated and analyzed using the Shimadzu Class-VP (Chromatography Laboratory Automated Software System (Tokyo, Japan). Plant samples, standard solutions, and mobile phases were filtered using a 0.45 micrometer pore size membrane filter (Vivascience AG, Hannover, Germany). The amount of phenolic contents in the plant sample was calculated as g kg -1 herb using external calibration curves, constructed for each pure phenolic standart. All determinations were carried out in triplicate and the results were presented as mean ± standard error.
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