Gradient HPLC analysis was done by using Shimadzu Prominence series UFLC system with a CBM-20A controller bus module, a LC-20 AD liquid chromatograph, a CTO-20A column oven and a SPD-20A UV-visible detector. UV-visible absorption was measured at 295 nm. 20 μL of sample were loaded in the solvent injection ratio: 95% solvent A – 5% solvent B (A = Milli-Q water/TFA 99.9:0.1 v/v; B = CH3CN/Milli Q water/TFA 90:9.9:0.1 v/v/v) onto a Jupiter C4 column (150 × 4.60 mm, 5 μm, 300 Å, Phenomenex) at a flow rate of 1 mL/min over 5 min. In a second step, samples were eluted by a gradient developed from 5 to 90% of solvent B in solvent A over 15 min. The concentration of solvent B was maintained over 5 min. Then, the concentration of solvent B was decreased to 5% over a period of 5 min to re-equilibrate the system, followed by additional 5 min at this final concentration. Before each sample measurement, a baseline was performed following the same conditions by loading Milli-Q water into the injection loop.
Lc 20ad liquid chromatograph
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu LC-20AD is a liquid chromatograph designed for the separation and analysis of chemical compounds. It features a high-precision dual-plunger pump for reliable solvent delivery, and is capable of operating at pressures up to 40 MPa. The LC-20AD is a core component of analytical instrument systems used in various industries for applications such as quality control, environmental testing, and pharmaceutical research.
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Lab products found in correlation
Prominence series uflc system, by Shimadzu (2 mentions)
Jupiter c4 column, by Phenomenex (1 mentions)
Cbm 20a controller, by Shimadzu (1 mentions)
C18 4.6 mm 50 mm column, by Shimadzu (1 mentions)
Sil 20a ht autosampler, by Shimadzu (1 mentions)
Rezex roa organic acid column, by Phenomenex (1 mentions)
Spd 20a uv vis detector, by Shimadzu (1 mentions)
Trypsin solution, by Thermo Fisher Scientific (1 mentions)
Dorzolamide, by Merck Group (1 mentions)
Ac 400f, by Bruker (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Sil 20a autosampler, by Shimadzu (1 mentions)
Kinetex phenyl hexyl column, by Phenomenex (1 mentions)
Lcms 8050 triple quadrupole tandem mass spectrometer, by Shimadzu (1 mentions)
Atlantis t3 column, by Waters Corporation (1 mentions)
Xcalibur, by Thermo Fisher Scientific (1 mentions)
Atlantis t3 guard column, by Waters Corporation (1 mentions)
Tsq ultra triple quadrupole mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Perchloric acid, by Fujifilm (1 mentions)
14 protocols using lc 20ad liquid chromatograph
1
Gradient HPLC Analysis of Samples
2
HPLC Analysis of Bacterial Metabolites
3
Quantifying Volatile Fatty Acids in Anaerobic Digestion
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The concentration of VFAs (acetate, formate, butyrate, propionate, and valerate) formed during anaerobic digestion was measured through High Pressure Liquid Chromatography (HPLC). Culture samples were centrifuged at 13,000 rpm for 3 min and filtered using 0.22 mm syringe filters. A Shimadzu LC-20AD liquid chromatograph (Shimadzu, Milton Keynes, UK) equipped with a Shimadzu SPD-20A UV/VIS detector, a Shimadzu SIL-20A HT auto sampler, and a CTO-10AS VP column oven was used. The column was eluted isocratically at a rate of 0.7 mL min−1 from an organic analysis column (Rezex ROA-Organic Acid column, Phenomenex, Torrance, CA, USA) with 5 mM H2SO4 at 55 °C, while the injection volume was 1 μL.
4
Synthesis and Characterization of Amphiphilic Copolymers
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Hydrophobic Dorzolamide and IMC were purchased from Sigma Aldrich (St. Louis, MO, USA). All solvents were used as received without further purification. The human dermal fibroblast cell line (HDFa) and the necessary supplies (antibiotic cocktail: penicillin and streptomycin; non-essential amino acids; trypsin solution; and foetal bovine serum—FBS) for the in vitro cytotoxicity assay were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Two copolymer samples were investigated in the present study. The PCL-g-P(NVCL-co-NVP) copolymers were obtained by a grafting-from technique in three steps, as previously reported [24 (link)]. The backbone was synthesised by ring-opening copolymerisation of ε-caprolactone (ε-CL) and α-chloro-ε-caprolactone (α-Cl-ε-CL). In the second step, a RAFT macroinitiator was prepared by the substitution of pendant chloro groups with a xanthate salt. Finally, RAFT-Madix copolymerisation of N-vinyl caprolactam (NVCL) with N-vinylpyrrolidone (NVP) was carried out. The obtained copolymers were analysed by 1H NMR (Bruker AC-400F, MA, USA) and size exclusion chromatography (Shimadzu LC-20AD liquid chromatograph, Japan), and the molecular characteristics of the two samples are provided inTable 4 .
Two copolymer samples were investigated in the present study. The PCL-g-P(NVCL-co-NVP) copolymers were obtained by a grafting-from technique in three steps, as previously reported [24 (link)]. The backbone was synthesised by ring-opening copolymerisation of ε-caprolactone (ε-CL) and α-chloro-ε-caprolactone (α-Cl-ε-CL). In the second step, a RAFT macroinitiator was prepared by the substitution of pendant chloro groups with a xanthate salt. Finally, RAFT-Madix copolymerisation of N-vinyl caprolactam (NVCL) with N-vinylpyrrolidone (NVP) was carried out. The obtained copolymers were analysed by 1H NMR (Bruker AC-400F, MA, USA) and size exclusion chromatography (Shimadzu LC-20AD liquid chromatograph, Japan), and the molecular characteristics of the two samples are provided in
5
HPLC Determination of Geraniol in Microparticles
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Both DL and EE were determined using HPLC (LC-20AD liquid chromatograph, Shimadzu, Japan) consisting of two LC-10ADvp pumps, a DGU-20A continuous degassing unit equipped with an SIL-20A autosampler and an SPD-20A UV–vis detector. Chromatographic separations were achieved by a Pronto SIL® RP-C18 (150 × 4.6 mm, 5 μm) column (type SC-150, Bischoff Chromatography, Berlin, Germany).
Data acquisition and processing were performed on a personal computer using LC solution Software, version 1.25 SP4 (Shimadzu, Kyoto, Japan). The detector was set at 210 nm and the temperature was maintained at 25 °C. The mobile phase consists of an isocratic mixture of acetonitrile and deionized water (50/50, v/v) at a flow rate of 1 mL/min [29 (link)]. The retention time obtained was 6.9 min for geraniol.
Briefly, a known amount of MM was dissolved in methanol to quantify the geraniol concentration. The DL and EE were calculated by the following equations:
Data acquisition and processing were performed on a personal computer using LC solution Software, version 1.25 SP4 (Shimadzu, Kyoto, Japan). The detector was set at 210 nm and the temperature was maintained at 25 °C. The mobile phase consists of an isocratic mixture of acetonitrile and deionized water (50/50, v/v) at a flow rate of 1 mL/min [29 (link)]. The retention time obtained was 6.9 min for geraniol.
Briefly, a known amount of MM was dissolved in methanol to quantify the geraniol concentration. The DL and EE were calculated by the following equations:
6
LC-MS/MS Quantification of Estradiol
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Drug quantification by LC–MS/MS was based on the principles of isotope dilution [28 (link)]. ORDX SD rats (n = 5–7) were treated with DHED (100 μg/kg, p.o.) and then euthanized 0.25, 0.5, 1, 2, 4, and 8 h after treatment. The hypothalami were dissected from the harvested brains, followed by tissue homogenization (20% w/v, pH 7.4 phosphate buffer) and adding 13C-labeled internal standard (IS) (13C6-E2, 100 pg) for E2 quantitation, as reported previously [16 (link),19 (link),28 (link)]. Liquid–liquid extraction was performed with 4 volumes of methyl tert-butyl ether. The organic layers obtained from the extractions were removed and evaporated under a nitrogen stream to yield samples for derivatization and subsequent LC–MS/MS analysis using electrospray ionization. Derivatization of the analyte and the IS was done by dansyl chloride [28 (link)]. The dansylated samples were centrifuged at 4500 rpm for 3 min, transferred to autosampler vials, sealed, and assayed using an LC–MS-8050 triple quadrupole tandem mass spectrometer connected to an LC-20AD liquid chromatograph (Shimadzu, Tokyo, Japan). Separation was performed according to our earlier publication using a Phenomenex (Torrance, CA, USA) Kinetex phenyl-hexyl column (50 mm × 2.1 mm i.d., 2.6 µm particles with 100 Ǻ pores), and quantification relied on selected-reaction monitoring [28 (link)].
7
Iris pallida Rhizome Extraction and HPLC Analysis
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A total of 50 mg of the rhizome of Sweetie Iris (Iris pallida) was accurately weighed into a 50 mL tube and extracted as shown in the extraction procedure. An aliquot of the extract after centrifugation was filtered through a 0.2 μm syringe filter before injection (10 µL). HPLC analysis was performed using an LC-20AD liquid chromatograph (Shimadzu, Kyoto, Japan). An Itertsil ODS 2 column (250 × 4.6 mm, 5 μm particles, GL Science Inc., Tokyo, Japan) was used. Mobile phases A and B were 0.1% phosphoric with water and MeOH, respectively. The column temperature was maintained at 30 °C, and the flow rate of the mobile phase was set at 0.5 mL min−1. The following multi-step gradient with different proportions of mobile phase B was applied: 0 min, 10% B; 5 min, 40% B; 10 min, 80% B; 20 min, 20% B and maintained until 30 min before injection (20 μL). The analysis was monitored using an SPD-M20A detector at 265 nm. Quantification was obtained by comparing the peak areas of the target compounds with the abundance of these compounds in the corresponding standards used in the calibration curve. All chemical analyses were performed in three replications.
8
GO Trapping Capacity Determination
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The GO trapping capacity was tested using the method described by Peng et al. (20 (link)) with some modifications. PD (derivatization agent, 20 mM) was freshly prepared in PBS (pH 7.4, 0.02 M). The sample was mixed with the peritoneal dialysis (PD) solution in the ratio of 1:1 and maintained at 37°C for 0.5 h. The derivative product was then injected into an high-performance liquid chromatography (HPLC) system equipped with an LC-20AD liquid chromatograph (Shimadzu, Japan) and a Diamonsil TM-C18 column (4.6 × 200 mm, 5 μm, Shimadzu Co., Kyoto, Japan).
9
HPLC Analysis of Phytochemicals
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HPLC experiments were carried out on a Shimadzu LC-20AD Liquid Chromatograph (SPD-M20A diode array detector, CBM-20A controller, SIL-20ACHT autosampler, DGU-20A5R degasser unit, CTO-20AC column oven) using a Kinetex 5 μm C-18 100A (150 mm × 4.6 mm) with a gradient of 0,01% trifluoroacetic acid in H2O (A) and acetonitrile (B) as follows: 0–5 min 25% B, 14 min 28% B, 15 min 70% B, 16 min 70% B, 16.5 min 25% B, and 20 min 25% B. The flow was 1.2 ml/min, column oven temperature was 55°C. Detection was carried out within the range of 190–800 nm. For quantification, chromatograms were integrated at 344 nm. The reference standards and the evaporated extracts were dissolved in MeOH, filtered through a PTFE syringe filter and injected in volumes of 5 or 10 μl. Calibration curves were established for all the four reference standards.
10
Quantitative Analysis of 5-Fluorouracil in Plasma
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