S. cerevisiae cell pellets from 5 ml cultures were extracted with MeOH, and the supernatant was extracted with nBuOH. Extracts were concentrated and analyzed by HPLC (Shimadzu SPD-20A system, YMC ODS-A column [4.6 id × 250 mm], MeOH—5 mM phosphate buffer (1% MeOH for 20 min followed by a gradient from 1 to 95% MeOH in 20 min), flow rate 0.3 ml/min, 296 nm.
Spd 20a system
Manufactured by Shimadzu
Sourced in Japan
The SPD-20A is a UV-Vis detector system designed for high-performance liquid chromatography (HPLC) analysis. It provides reliable and precise absorbance measurements across a wide wavelength range. The core function of the SPD-20A is to detect and quantify the components in a liquid sample as they elute from the HPLC column.
Automatically generated - may contain errors
Lab products found in correlation
Ods a column, by Shimadzu (1 mentions)
Infinite f200 pro microplate reader, by Tecan (1 mentions)
Jnm eca600, by JEOL (1 mentions)
Jnm ecs400, by JEOL (1 mentions)
Jms t100td, by JEOL (1 mentions)
Auto gamma system arc 7010b, by Hitachi (1 mentions)
Avance 3 500 mhz, by Bruker (1 mentions)
1260 infinity 2 system, by Agilent Technologies (1 mentions)
6530 accurate mass q tof, by Agilent Technologies (1 mentions)
P 2000 polarimeter, by Jasco (1 mentions)
J sphere ods h80, by YMC (1 mentions)
400 mhz nmr spectrometer, by Agilent Technologies (1 mentions)
4 protocols using spd 20a system
1
Yeast Metabolite Extraction and HPLC Analysis
2
Analytical Techniques for Chemical Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Auto Gamma System ARC-7010B (Hitachi, Ltd., Tokyo, Japan) recorded radioactivity. Thin-layer chromatography (TLC) on silica plates 60 F254 (Merck, Darmstadt, Germany) was used to monitor the reactions. High-performance liquid chromatography (HPLC) of SPD-20A system (Shimadzu Corp., Kyoto, Japan), using a Cosmosil® 5SL-II (20 ID × 250 mm) column (Nacalai Tesque, Inc., Kyoto, Japan). Nuclear magnetic resonance (NMR) spectroscopy was conveyed on JNM-ECS 400 and JNM-ECA 600 (JEOL Ltd., Tokyo, Japan). Direct analysis in real-time mass spectrometry (DART-MS) and electrospray ionization mass spectrometry (ESI-MS) were conducted on JMS-T100TD (JEOL Ltd., Tokyo, Japan). The optical density in 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H tetrazolium monosodium salt (WST-8) assay was conducted on Infinite® F200 Pro microplate reader (TECAN, Männedorf, Switzerland).
3
NMR, Optical Rotation, and HPLC Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
General procedures: The NMR spectra were recorded on a Bruker Avance III 500 MHz. Optical rotation was recorded on a Jasco P2000 polarimeter. HR-ESI-MS was acquired on an Agilent 6530 Accurate-Mass QTOF. Column chromatography (CC) was performed using silica gel (40-63 µm) , sephadex LH-20, or reversed phase C-18 (RP18, 150 µm) as adsorbents. TLC was performed on pre-coated plates. Semi-preparative HPLC was performed on an Agilent 1260 infinity II system including binary pump, autosampler, DAD detector, fraction collector, and equipped with YMC J'sphere ODS-H80 (20×250 mm, 4µm) column. Mobile phase was an isocratic system of acetonitrile/water at flow rate of 3 mL/min. HPLC chromatogram was carried out on the Shimadzu SPD-20A system (C18 column; 250 mm, 4.6 mm, 5µm) (Shimadzu Co., Ltd., Kyoto, Japan) . A mixture of methanol and water solution was used as the mobile phase.
4
Characterization of Radiolabeled Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All the chemicals were purchased from commercial sources and used without further purification. 1H NMR spectra were recorded on a Varian 400‐MHz NMR spectrometer. 1H NMR chemical shifts were determined relative to CD3CN at δ 1.94 ppm. All radioactive reactions and products were analyzed with high‐performance liquid chromatography (HPLC) using a Shimadzu SPD‐20A system and LabSolutions 5.85 software (Shimadzu Corporation, Japan) with a Grace Smart C18 column (5 μ 4.6 × 250 mm), MeCN/H2O/TFA 30:70:0.1 or an Alltima C18 (5 μ 4.6 × 250 mm), MeCN/H2O/TFA 20:80:0.1 as eluent and a flow of 1 ml/min. UV active compounds were detected at 254 nm. Radioactive products were identified by comparison with the unlabeled reference compounds. Radioactivity was quantified with a Veenstra VDC‐304 dose calibrator.
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!