All reagents and solvents from commercial sources were of the highest purity available and were used as received. Elemental analyses were obtained at the microanalysis service of the Milano CIMA department. CD spectra were obtained with a Jasco J500 spectropolarimeter (JASCO Corporation, Tokyo, Japan); spectra were recorded in the range 300–700 nm, at 50 nm/min, with three scans acquired for each spectrum, and 0.2 nm resolution. NMR spectra were recorded on a Bruker AVANCE 400 spectrometer (Bruker Italia, Milan, Italy), operating at 9.37 T. Data acquisition and processing were performed using a standard Bruker software package (Topspin 1.3). UV-vis spectra were recorded on an Agilent 8453 spectrophotometer. HPLC analyses were performed on a Jasco HPLC instrument (JASCO Corporation, Tokyo, Japan) equipped with two PU-1580 pumps and an MD-1510 diode array detector (working range: 195–659 nm) or on a Shimadzu HPLC instrument (Shimadzu Corporation, Kyoto, Japan) equipped with two LC-20AD pumps and an SPD-M20A diode array detector (working range: 190–800 nm). Mass spectra were recorded using a Thermo-Finnigan LCQ ADV MAX spectrometer (Thermo-Finnigan, San Jose, CA, USA).
Hplc instrument
Manufactured by Jasco
Sourced in Japan
The HPLC instrument is a high-performance liquid chromatography system used for the separation, identification, and quantification of various chemical compounds. It is a versatile analytical tool that employs high pressure to move a liquid mobile phase containing the sample through a stationary phase within a column, allowing for the separation and detection of the individual components in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Agilent 8453 spectrophotometer, by Agilent Technologies (1 mentions)
Avance 400 spectrometer, by Bruker (1 mentions)
Redisep column, by Teledyne (1 mentions)
Superose 6 column, by GE Healthcare (1 mentions)
Refractive index detector, by Jasco (1 mentions)
Cation h refill guard column, by Bio-Rad (1 mentions)
As 2057 plus intelligent sampler, by Jasco (1 mentions)
Fp 2020 plus fluorescence detector, by Jasco (1 mentions)
Pu 2089, by Jasco (1 mentions)
8 protocols using hplc instrument
1
Analytical Characterization of Novel Compounds
2
Chromatographic Purification of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following organic synthesis, chromatographic purification of substances was carried out on either a Combiflash Rf+ flash chromatographic instrument (TELEDYNE Isco, Lincoln, NE, USA) equipped with DAD-ELS detection using commercially available RediSep columns (TELEDYNE Isco, USA), or on an Armen Spot Prep II preparative HPLC purification system (Gilson, Middleton, WI, USA) equipped with a dual-wavelength UV-VIS detector. Flash chromatographic purifications were performed on 4–24 G silica columns with adequately chosen eluent ratios of dichloromethane—methanol. For preparative HPLC separations, a Phenomenex Luna® 5 µm Silica (2) 100 Å 250 mm x 21.2 mm column (Phenomenex Inc., Torrance, CA, USA) was used, typically in isocratic elution mode with adequately chosen eluent ratios of cyclohexane—isopropanol. In general, the applied flow rates were 15 ml/min, and the wavelengths of detection were 210 andd 250 nm. Purity of the isolated compounds was determined on a Jasco HPLC instrument (Jasco International Co. Ltd., Hachioji, Tokyo, Japan) equipped with LC capillary cables proper for analytical purposes. The analysis was carried out on a 250 mm x 4.6 mm analogue of the preparative silica column using the peak area % data of the PDA chromatogram recorded between 210 and 410 nm. During the analytical-scale HPLC measurements, the applied flow rate was a constant 1 ml/min.
3
Identifying Polyamine-Protein Conjugates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The polyamine derivatives that covalently bound to the proteins were identified in the TCA-insoluble fractions obtained from the TGase radiolabeled assay, as described by Beninati et al. (2013 (link)), except that the Jasco HPLC instrument was used for chromatographic separation.
4
Size Exclusion Chromatography of Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Size exclusion chromatography (SEC-HPLC) was carried out using a Superose-6 column (GE Healthcare). The protein concentration was 10–30 μM, a volume of 50 μL was typically injected, and the running buffer was 20 mM Tris–HCl, 100 mM NaCl, 1 mM EDTA, pH 7.0. The experiment was carried out at room temperature (∼ 25 °C) at a 0.5 mL min−1 flow rate. A JASCO HPLC instrument was used. It was equipped with an automatic injector, a quaternary pump and a UV–VIS UV-2075 (the elution was monitored at 280 nm).
5
Fermentation Assay for Sugar Metabolism
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fermentation assays were carried out as described previously with slight modifications [36 (link), 37 (link)]. In brief, cells were pre-cultured to the stationary phase at 30 °C in 50 mL YPD medium supplemented with an appropriate antibiotic and washed once with YPD85X35 medium. Cells (initial cell density of 20 ODA600 unit) were then inoculated into 70 mL fresh YPD85X35 media in 100-mL flasks capped with rubber stoppers with inserted syringe needles to release CO2. The flasks were also fitted with T-shape stopcocks for ventilation and sampling. During fermentation, 500 µL from each culture medium was collected via the sampling port at specified intervals (0, 1, 3, 6, 12, 24, 36, 48, 60 and 72 h). Samples were stored at − 80 °C until assessment by HPLC. All fermentation assays were performed in triplicate unless otherwise stated. To determine the concentration of sugars and fermentation products such as glucose, xylose, xylitol, glycerol, acetate and ethanol in the culture media, specimens were diluted fivefold with deionized water and analyzed by an HPLC instrument coupled to a refractive index detector (Jasco, Tokyo, Japan) and fitted with an Aminex HPX-87H column and Cation H Refill Guard column (Bio-Rad, Hercules, CA). HPLC assays were performed at 65 °C with 5 mM H2SO4 buffer as the mobile phase, a flow rate of 0.6 mL/min, and an injection volume of 25 µL.
6
Analytical Method for Phenolic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phenolic acid analysis was performed following a previous method after minor modifications [19 (link)]. A Jasco HPLC instrument (Tokyo, Japan), equipped with a quaternary gradient pump Jasco PU-2089, an autosampler Jasco AS-2057 Plus Intelligent Sampler and two detectors, a Jasco UV/Vis MD-910 PDA detector and a Jasco FP-2020 Plus Fluorescence detector, was used. The column was a C18 Poroshell 120 (Agilent technologies, Santa Clara, CA, USA), 2.7 μm, (4.6 mm × 150 mm), operating at 35 °C with a flow of 0.8 mL/min. Elution solvents were 2% acetic acid in HPLC grade water (Eluent A) and 2% acetic acid in HPLC grade acetonitrile (Eluent B). Gradient elution was as follow: from 98% to 95% A in 10 min, 95% to 90% A in 7 min, 90 to 82% A in 6 min, 82% to 80% A in 3 min, 80% to 70% A in 3 min, 70% to 50% A in 3 min, 50% to 0% A in 4 min and 98% A in 1 min. Quantification of phenolic compounds was carried out using an external calibration curve obtained by injecting solutions of standard compounds at known concentrations and plotting peak areas vs. concentrations. The amount of tartrate esters of caffeic, coumaric and ferulic acids and Grape reaction Product GRP were expressed as the respective hydroxycinnamic acid.
7
Chloroform-Based Cardiac Bioanalysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Heart chloroform extracts were evaporated under the stream of nitrogen. The evaporated sample was then dissolved in approximately 0.5 ml of 80 % acetonitrile in water. 50 µl of this solution was applied on C-18 column (4x250 mm, 5 μm) attached to a JASCO HPLC instrument equipped with fluorescence detector. The detector was set at the excitation of 360 nm and the emission of 410 nm. The chromatography was run in the linear gradient mode between 20-80 % acetonitrile in water during 40 min. The flow rate was 0.7 ml min -1 , the column was thermostated at 30 °C.
8
Mitochondrial Lipid Extraction and HPLC Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondria chloroform extracts were evaporated under the stream of nitrogen. The evaporated sample was then dissolved in approximately 0.5 ml of 80 % acetonitrile in water. 50 µl of this solution was applied on C-18 column (4x250 mm, 5 μm) attached to a JASCO HPLC instrument equipped with fluorescence detector. The detector was set at the excitation of 357 nm and the emission of 430 nm. The chromatography was run in the linear gradient mode between 20-80 % acetonitrile in water during 40 min. The flow rate was 0.7 ml min -1 , and the column was thermostated at 30 °C.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!