The remaining silica gel of the bioactive band was scraped from the TLC plate and then eluted with 20 mL acetone. The acetone eluate was dried under vacuum, and the residues were dissolved in 0.5 mL HPLC grade methanol. After centrifuged at 12,000 rpm for 5 min, 200 µL of the methanol solution was subjected to HPLC (Shimadzu LC-20AT) equipped with Agilent ZORBAX SB-C18 column (9.4 × 250 mm, 5 µm). The mobile phase was acetonitrile-water solution at the flow rate of 2 ml/min in gradient elution method: 10% acetonitrile for 0–5 min; 10% to 70% acetonitrile for 5–45 min; 70% to 90% acetonitrile for 45–55 min; 90% acetonitrile for 55–60 min. The detection wavelength used was 215 nm. For identification of the bioactive peaks, the fraction collected per minute and yielded 58 fractions (0–3 min as fraction 1). All fractions were dried under vacuum and dissolved in 50 μL methanol. Twenty microliters of each fraction was screened against methicillin-resistant Staphylococcus aureus ATCC 33591 by the disc diffusion method. After incubated at 37 °C for 24 h, the diameters of the inhibition zones of bands were measured by Vernier caliper.
Lc 20at
Manufactured by Agilent Technologies
Sourced in Japan
The LC-20AT is a high-performance liquid chromatography (HPLC) system manufactured by Agilent Technologies. It is a core component of the company's analytical instrumentation portfolio. The LC-20AT is designed to provide precise and reliable liquid chromatography analysis, meeting the demands of a wide range of applications in various industries.
Automatically generated - may contain errors
Lab products found in correlation
Spd m20a, by Agilent Technologies (2 mentions)
Zorbax sb c18 column, by Agilent Technologies (1 mentions)
Hc c18 column, by Agilent Technologies (1 mentions)
Avance 3 600 mhz, by Bruker (1 mentions)
Avance 3 500 mhz, by Bruker (1 mentions)
Eclipse xdb c18, by Agilent Technologies (1 mentions)
Eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Agilent 1260 infinity, by Agilent Technologies (1 mentions)
Hplc system, by Shimadzu (1 mentions)
Aa700, by PerkinElmer (1 mentions)
8 protocols using lc 20at
1
Bioactive Compound Extraction and HPLC Analysis
2
Efficient 5-FU Loading in MOFs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5-Fluorouracil (5-FU) was selected as the model drug, and the carrier with the highest loading capacity of 5-FU was subjected to further analysis. 5-FU (20 mg) and MOFs (10 mg) in 10 mL of ethanol were dispersed by sonication and stirred at 25 °C for 6 h. Then, solvent removal was performed under reduced pressure, and the obtained products were washed with ethanol for three times. The drug-loading efficiency of MOFs was determined by high performance liquid chromatography (HPLC; Agilent LC-20AT).
3
Tetracycline Degradation Experiments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TC degradation experiments were conducted in 500 mL conical flasks containing 250 mL of TC solution (50 mg L−1) and a certain quantity of the catalyst (0.25 g) at 30 °C to maintain the same reacting temperature and mixing rate. All batch experiments were implemented in an air bath shaker at 180 rpm. At a given time interval, 0.5 mL of the reaction solution was taken, and then the TC concentration was detected with a 0.22 μm cellulose acetate membrane. The concentration of TC was analyzed by high performance liquid chromatography (HPLC, Shimadzu LC-20AT, Japan) with Agilent HC-C18 column (5 μm, 150 × 4.6 mm) at λ = 356 nm. The mobile phase was the organic phase (acetonitrile : methanol = 2 : 1, v = 0.28 mL min−1) and the aqueous phase (0.01 M, oxalic acid, v = 0.56 mL min−1) at an isocratic flow rate of 0.84 mL min−1. After a cycle of TC adsorption and degradation, the used catalyst was collected by centrifugation, washed with deionized water, and vacuum dried at 60 °C.
4
Lignin Structural Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The samples for HSQC and DOSY analysis were prepared by dissolving 30 mg lignin or the leftover material in to 0.6 mL DMSO-d6. The samples were sonicated at room temperature for 1 h to ensure dissolution. The HSQC experiments were conducted on a Bruker Avance III 600 MHz spectrometer at room temperature. The DOSY experiments were conducted on a Bruker Avance III 500 MHz spectrometer, and the temperature was set and maintained at 348 K. The data analysis was performed using the Bruker Dynamics Center 3.7 according to the procedure reported in the literature. The HPLC analysis was conducted on an Agilent LC-20AT liquid chromatograph with a C18 chromatographic column and a differential refraction detector (RID-20C). After the reaction, the reaction mixture was filtered through a 0.22 μm PTFE syringe filter, and the filtrate was used for HPLC analysis. The mobile phase was methanol aqueous solution (MeOH : H2O volume ratio of 55 : 45) with a flow rate of 0.8 mL min–1, and the temperature was kept at 50 °C. The trace was collected for 80 min.
5
Quantitative Analysis of Fungal Ophiobolins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells and dodecane phase were separated by centrifugation at 12,000 rpm for 5 min. The extracellular excreted ophiobolins were captured by dodecane, and diluted with ethyl acetate to an appropriate concentration for detection. The cells were suspended in acetone to extract the ophiobolins not excreted, then mixed on a vortex for 5 min and ultrasound extracted for 10 min, followed by centrifugation to collect the supernatant for analysis. Liquid chromatography (SHIMADZU LC-20 AT) equipped with an Eclipse XDB-C18 column (250 × 4.6 mm, 5 μm, Agilent Technologies, Germany) was used for the quantitative analysis of ophiobolins. The quantification of yields is based on the integration of UV signals. The mobile phase was 100% acetonitrile, with a flow rate of 1 mL/min for 30 min, and the column temperature was 25 °C. Detection wavelengths were set at 190 nm for OphF and 240 nm for OphU and 5-hydroxy-21-formyl-Ophiobolin F. The retention times of OphF, 5-hydroxy-21-formyl-Ophiobolin F, and OphU were 17.5, 5.5, and 4.7 min, respectively (Additional file 1: Figure S4 ). The ophiobolins isolated by semi-preparative HPLC (Agilent 1260 Infinity, Agilent Technologies, Germany) equipped with an Eclipse XDB-C18 (250 × 9.4 mm, 5 μm, Agilent Technologies, Germany). The mobile phase was 100% acetonitrile, with a flow rate of 3 mL/min for 30 min, and the column temperature was 25 °C.
6
HPLC Analysis of Compound Mixture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A previously reported method was slightly modified for HPLC analysis (18 ). The Shimazu HPLC system consisted of a pump (LC-20AT), a diode array detector (DAD, SPD-M20A), and a 300SB-C18 column (Agilent 250×4.6 mm). The sample solution was filtered through a 0.45-µm nylon membrane filter (Millipore), and 10 µL was injected into the liquid chromatographer. The mobile phase was methyl alcohol–water–phosphoric acid, with a ratio of 500:500:0.4 (v/v/v). The flow rate was 1 ml/min, and the eluate absorbance was monitored at 370 nm using a scanning range of 200–600 nm.
7
Hydrolysis and HPLC Analysis of EE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A previously reported method was slightly modified for hydrolysis and high-performance liquid chromatography (HPLC) analysis (28 (link)). EE was dissolved in ethanol–hydrochloric acid–water (7/2/1, v/v/v) solution and at 75°C by ultrasound (40 kHz) for 60 min. Then, the supernatant was used in the following analysis.
HPLC was conducted by using a Shimadzu HPLC system (Shimadzu, Japan), which consisted of a pump (LC-20AT), diode array detector (DAD, SPD-M20A), C18 column (Agilent 250 × 4.6 mm), and LC-solution system manager program. The mobile phase comprised methyl alcohol–water–acetic acid in a ratio of 500/500/0.4 (v/v/v). The flow rate was 1 mL/min, and the eluate absorbance was monitored at 370 nm using a scanning range of 200–600 nm. The injection volume of the extract was 10 μL.
HPLC was conducted by using a Shimadzu HPLC system (Shimadzu, Japan), which consisted of a pump (LC-20AT), diode array detector (DAD, SPD-M20A), C18 column (Agilent 250 × 4.6 mm), and LC-solution system manager program. The mobile phase comprised methyl alcohol–water–acetic acid in a ratio of 500/500/0.4 (v/v/v). The flow rate was 1 mL/min, and the eluate absorbance was monitored at 370 nm using a scanning range of 200–600 nm. The injection volume of the extract was 10 μL.
8
Quantitative Analysis of TC, Cu2+, and Cd2+ in Supernatants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentration of TC in the supernatants was determined by high performance liquid chromatography (Shimadzu, LC-20 AT, Japan) equipped with an Agilent Eclipse XDB-C18 reversed-phase column (4.6 × 150 mm, 5 μm, Supelco, USA) with column temperature at 30 °C. The mobile phase was 20 : 80 (v/v) of acetonitrile and 0.01 M oxalic acid at a flow rate of 1.0 mL min−1. TC was analyzed by a UV detector at 360 nm. The concentration of Cu2+ and Cd2+ in the supernatants was determined using an atomic absorption spectrophotometer (PerkinElmer, AA700, USA).
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!