Chromatographic separations of amino acids were performed on a Shimadzu HPLC system consisting of two LC-20AD isocratic pumps, a DGU-14A degasser, an SIL-10AD autosampler and a fluorescence detector (RF-535) (Shimadzu, Tokyo, Japan). The analytical column was a reversed phase LC-C18 DB (250 × 4.6 mm, 5.0 μm, Supelco (Bellefonte, United States). LC-solution® software version 1.25 SP4 was utilized for hardware control and data manipulation. All separations were accomplished using a binary gradient elution program. The mobile phases A and B were 20 mM phosphate buffer (pH 2.8) and methanol, respectively. Their initial ratio was 40% v/v of B and the flow rate was set at 0.7 mL min−1. The ratio of the mobile phase B was linearly increased to 65% in 15 min and kept constant for 9 min. Then, it was further increased to 80% in 1 min, while the flow was decreased at 0.6 mL min−1 and stayed constant for up to 30 min. Then, it reverted to its initial conditions (40% B and flow 0.7 mL min−1) in 5 min and kept constant for up to 40 min to obtain reproducible separations. The injection volume was set at 20 μL (for the analysis of the product) and 100 μL (to determine the LOD value on the chocolate substrate). The column was thermostated at 25 °C. Amino acid-OPA derivatives were detected spectrofluorimetrically at_λex/λem = 340/455 nm.
Dgu 14a degasser
Manufactured by Shimadzu
Sourced in Japan
The DGU-14A degasser is a laboratory equipment that removes dissolved gases from liquid samples. It is designed to enhance the performance of high-performance liquid chromatography (HPLC) systems by preventing the formation of air bubbles and ensuring a stable baseline. The DGU-14A degasser operates by applying a vacuum to the liquid samples, effectively removing dissolved gases such as oxygen and nitrogen.
Automatically generated - may contain errors
Lab products found in correlation
Hplc system, by Shimadzu (4 mentions)
Scl 10a vp system controller, by Shimadzu (3 mentions)
Rf 535, by Shimadzu (2 mentions)
Sil 10ad autosampler, by Shimadzu (2 mentions)
Lc 20ad isocratic pumps, by Shimadzu (2 mentions)
Spd m20a, by Shimadzu (2 mentions)
Scl 10a system controller, by Shimadzu (2 mentions)
Lc 10at pump, by Shimadzu (2 mentions)
Cto 10avp column oven, by Shimadzu (2 mentions)
Sil 10af autosampler, by Shimadzu (2 mentions)
Lc 10atvp pump, by Shimadzu (1 mentions)
Avance 400, by Bruker (1 mentions)
Class vp software, by Shimadzu (1 mentions)
Spd m10avp diode array, by Shimadzu (1 mentions)
Sil 10advp auto injector, by Shimadzu (1 mentions)
Ascentis express 90a c18 column, by Merck Group (1 mentions)
Lc 10advp solvent delivery system, by Shimadzu (1 mentions)
Zorbax sb c18, by Agilent Technologies (1 mentions)
Spd 10av uv vis detector, by Shimadzu (1 mentions)
Sil 10ad auto injector, by Shimadzu (1 mentions)
13 protocols using dgu 14a degasser
1
HPLC Analysis of Amino Acids
2
NMR and HPLC Analysis of Compounds
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1H, 19F and 31P-NMR spectra and 2D NMR experiments were recorded on Bruker Avance 400 or 500 MHz instruments using D2O as solvent at 25 °C or deuterated borate buffer, pH 7.2 (pD 7.62).
HPLC measurements were performed on a Shimadzu HPLC system composed of a SPD-10UV-Vis detector, a LC-10AT VP pump with a DGU-14A degasser and a C18 Supelco HPLC column [(L × OD) = 25 cm × 4.6 mm, with 5 μm particle size]. Injection volumes were 20 μL. The detection wavelength was set at λ = 254 nm. The mobile phase was acetonitrile: water (75:25, v/v) at a flow rate of 0.5 mL/min.
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1H, 19F and 31P-NMR spectra and 2D NMR experiments were recorded on Bruker Avance 400 or 500 MHz instruments using D2O as solvent at 25 °C or deuterated borate buffer, pH 7.2 (pD 7.62).
HPLC measurements were performed on a Shimadzu HPLC system composed of a SPD-10UV-Vis detector, a LC-10AT VP pump with a DGU-14A degasser and a C18 Supelco HPLC column [(L × OD) = 25 cm × 4.6 mm, with 5 μm particle size]. Injection volumes were 20 μL. The detection wavelength was set at λ = 254 nm. The mobile phase was acetonitrile: water (75:25, v/v) at a flow rate of 0.5 mL/min.
3
Quantifying 5-Fluorouracil Residues on Surfaces
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Surface wipe samples were taken from the surface of the cabinets or isolators (where the drugs were prepared) before and after cleaning, as recommended by the international standards. The method was adopted from previous works (Ziegler et al., 2002[40 (link)]). The samples from surfaces were taken by wiping thoroughly the surface with sterile gauze (10 cm×10 cm) which had been wetted with 5 ml of 30 mM sodium hydroxide. All samples were collected into the separate sealed plastic boxes and kept at -18 °C before the analysis. The formic acid with the PH of 3 (5 ml) was added to each sample. The prepared samples were then sonicated and injected in to the HPLC. Analyses were carried out with the HPLC system model SPDM-10ADvp from Shimadzu (Kyoto, Japan) consisting of LC-10Advp binary pumps, a SCL-10Avp controller, a SPD-M10Avp PDA detector and a DGU-14A degasser model and operated with Class-VP software (Shimadzu Scientific instruments, Inc.). Separations were done on a C18-M51002546 250 × 4.6 mm column from Hector (Venture Town, South Korea). The mobile phase used for chromatography was formic acid with the PH of 3, with a wavelength of 200 and flow rate of 1.4. The selectivity of the method and accurate recognition of the substance (5-FU) was verified by the UV spectra.
4
HPLC Analysis of Herbal Compounds
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For HPLC, we used the Shimadzu LC-10ATvp liquid chromatograph with DGU-14ALvp, DGU-14A degasser, SIL-10ADvp auto injector, SCL-10Avp system controller, SPD-M10Avp diode array detector, and LCsolution ver. 1.25 software (Shimadzu, Kyoto, Japan). The Ascentis Express 90A C18 column (4.6 mm × 25 cm; 5 µm; Supelco) was used for the HPLC analysis of gentiopicrin, geniposide, astilbin, berberine chloride, and baicalin. The mobile phase consisted of water and 0.05% trifluoroacetic acid–acetonitrile–methanol under the following gradient program: 0–55 min, 90 : 5:5 ⟶ 50 : 5:45 linear gradient; 55–56 min, 50 : 5:45 ⟶ 0 : 0:100 linear gradient; 56–60 min, 0 : 0:100 isocratic; 60–61 min, 0 : 0:100 ⟶ 90 : 5:5 linear gradient; and 61–65 min, 90 : 5:5 isocratic. The flow rate was 1.0 mL/min, and 10 μL was injected into the column. The column temperature was maintained at 40°C. Chromatograms were recorded at 230 nm.
Coptisine, palmatine, and berberine chloride were determined following the assay method recommended in Taiwan Herbal Pharmacopeia, Third Edition, by using the Purospher STAR RP-18e column: elution buffer, 45 : 55 acetonitrile:H2O (containing monopotassium phosphate, 3.4 g/L and sodium lauryl sulfate, 1.7 g/L); flow rate, 1.0 mL/min; column temperature, 40°C; and detection wavelength, 345 nm.
Coptisine, palmatine, and berberine chloride were determined following the assay method recommended in Taiwan Herbal Pharmacopeia, Third Edition, by using the Purospher STAR RP-18e column: elution buffer, 45 : 55 acetonitrile:H2O (containing monopotassium phosphate, 3.4 g/L and sodium lauryl sulfate, 1.7 g/L); flow rate, 1.0 mL/min; column temperature, 40°C; and detection wavelength, 345 nm.
5
HPLC Analysis of Mulberry Root Bark Extract
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Dried ground root bark of M. alba (10 g) was soaked exhaustively with cold water (0.5 L) for 1 h. Subsequently the material was decocted for 20 min in a pot with a lid. After filtration through cotton wool, the decoction procedure was repeated with the remaining plant material. The filtrated decoctions were combined (MAD) and an aliquot was used for HPLC analyses. HPLC of MAE, MAF, and MAD (Supplementary Fig. 1 ) was performed on a Shimadzu device consisting of an LC-10ADVP solvent delivery system, a FCV-10ALVP low-pressure gradient flow control valve, an SCL-10AVP system controller, a DGU-14A degasser, and a SPD-M20A photodiode array (PDA) detector. LC-parameters: stationary phase: Agilent Zorbax SB-C18 3.5 μm (150 × 4.6 mm); temperature: 25 °C; mobile phase: A = water; B = acetonitrile; flow rate 1.0 mL/min; PDA detection wavelength: 205 nm; injection volume: 10 μL; Separations were performed by gradient elution (70/30 A/B in 5 min to 55/45 A/B, then within 15 min to 45/55 A/B, and within 2 min to 2/98 A/B), followed by a 10 min column wash (2A/98B) and a re-equilibration period of 10 min. All chemicals and solvents used were analytical grade.
6
Bromethalin Quantification by HPLC
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The HPLC used consisted of an SIL-10 AD Auto-injector, LC-10AD dual pump solvent delivery module, SCL-10A system controller, a DGU-14A degasser, and an SPD-10 AV UV-Vis detector (all are products of Shimadzu Corporation, Kyoto, Japan). Data collection and processing was done using Class-8000 LC/MS software, also from Shimadzu Corporation. Separation was done using Zorbax SB-C18 Rapid Resolution HT column 4.6 cm X 50 mm, 1.8 micron. The injected volume was 60 μl, UV detector set at 261 nm and the eluent was methanol-water (3:1) at a combined flow rate of 0.6 ml/minute. The retention times of desmethyl-bromethalin and bromethalin were 9.1 minutes and 21.4 minutes, respectively.
7
Nanoparticle Drug Loading Efficiency
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The loading efficiency and loading capacity was determined as follows. In brief, 10 mg of lyophilized NP was dissolved in 5 ml of DMSO and sonicated for 15 min. The organic solution was centrifuged and the supernatant was used to calculate the amount of drug loaded. The drug loading was quantified using HPLC method. The HPLC system (Shimadzu, Kyoto, Japan) consisted of LC-10AT pump, a SPD-10A UV/Vis detector and a DGU-14A degasser model. The flow rate was maintained at 1 ml/min. The wavelength of detection was 254 nm. 50 mM of KH 2 PO 4 (pH 5.0) was used as a mobile phase.
8
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.
9
Analytical Characterization of Organic Compounds
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All reagents were purchased from commercial suppliers and were used without further purification. Prominence HPLC system (Shimadzu, Kyoto, Japan), equipped with a Shimadzu DGU-14 A degasser, LC-20 A dual piston pump, CTO-10 ASVP column oven and SPD-MI20A PDA detector Sıl-20ACXR autosampler and a stainless-steel GL Science Inertsil ODS-3 (4.6 250 mm) column. Solvents (acetonitrile and water, 7:3) for the separation obtained compounds were isocritically mixed at a flow rate of 0.25 mL/min and injection were performed by volume of 10 µL. IR spectra was recorded on an IR Affinity-1S Infrared spectrophotometer (Shimadzu, Tokyo, Japan). Melting points (M.p.) were determined using the Mettler Toledo-MP90 Melting Point System and were uncorrected. 1H NMR and 13C NMR spectra was recorded on a Bruker Fourier 300 (Bruker Bioscience, Billerica, MA, USA) respectively, in DMSO-d6. (Bruker, Billerica, MA, USA), respectively. MS studies were performed on an LCMS-8040 tandem mass system (Shimadzu, Tokyo, Japan).
10
HPLC Analysis of Ethanolic Extracts
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The ethanolic extract was diluted in methanol until a final concentration of 10 mg mL -1 , and then submitted to HPLC analysis using a Shimadzu SCL-10AVP equipped with DGU-14A degasser, LC-10AD binary pump, CTO-10AS oven, SIL-10AF auto-injector and SPD-M10A diode array detector. The analyses were performed on a reverse-phase LiChrospher-100 RP-18 Merck column (250 mm × 4.6 mm i.d.; 5 μm particle size; Darmstadt, Germany) equipped with a Merck guard column (10 mm × 4.0 mm i.d.), packed on the same packing material, and placed before the inlet of the analytical column. A gradient of ultrapure water acidified with glacial acetic acid (pH = 3.0) (A) and acetonitrile (B) was used as follows: 10% (0 min), 90% (80 min) and 10% (90 min); nonpolar compounds: H 2 O -90% (0 min), 10% (80 min) and 90% (90 min) at a rate of 1.0 mL/min, 40 °C (λ 290 nm), oven temperature set at 50 °C. Detection was performed at 220 nm and 340 nm. Chromatograms were visualized and manipulated using the software Class-VP.
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