Sacculi (or glycan chains) were digested with cellosyl, boiled to deactivate enzyme and then reduced with sodium borohydride as previously described5 (link). The resulting soluble material was separated by reverse-phase high-performance liquid chromatography (RP-HPLC) using a Hypersil aQ C18 column (3 μm, 2.1 by 200 mm) coupled to an Agilent 6500 series quadrupole time of flight mass spectrometer (Q-TOF LC–MS). Flow rate was 0.15 ml/min. Buffer A was water and buffer B was acetonitrile, both containing 0.1% (v:v) formic acid. The gradient was 0 to 15% buffer B over 35 min.
Aq c18 column
Manufactured by Agilent Technologies
Sourced in United States
The AQ-C18 column is a reverse-phase high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a wide range of polar and non-polar compounds. The column features a C18 stationary phase that provides high-quality chromatographic performance and selectivity.
Automatically generated - may contain errors
5 protocols using aq c18 column
1
Glycan Chain Analysis by RP-HPLC-QTOF
2
Quantification of Ergosterol in Aspergillus fumigatus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total ergosterol in the A. fumigatus strains was extracted using a previously described protocol (Alcazar-Fuoli et al., 2008 (link)). Briefly, mycelia were cultured for 18 h in 100 ml of liquid MM media. Subsequently, they were harvested, dried and ground to a fine powder. Two hundred milligrams of the ground mycelia were treated with 3 ml of 25% alcohol potassium hydroxide solution (3:2 methanol:ethanol) and mixed by vortexing for 1 min. After the mixture was incubated in an 85 °C water bath for 1 h, the ergosterol was treated with 1 ml of sterile distilled water and 3 ml of pentane followed by vigorous vortexing for 3 min. The upper pentane layer was transferred to a clean glass tube and evaporated in a fume hood at room temperature. The dried-down samples were re-dissolved in 1 ml of methanol and syringe-filtered through 0.2 μm-pore-size filters. Total ergosterol was analyzed using HPLC (Agilent Technologies) and detected at 280 nm using an AQ-C18 column (250 mm by 4.6 mm, 5 μm). Elution was conducted at a flow rate of 1 ml min-1 with a mobile phase containing water and methanol eluent (100% HPLC grade).
3
Quantifying Ergosterol in A. fumigatus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The extraction of total ergosterol from A. fumigatus mycelia was performed with minor modifications as described previously (Long et al., 2017 (link)). Briefly, approximately 1 × 108 spores were incubated in 100 ml liquid media with relevant concentrations or drugs for 24 h at 37°C at a speed of 220 rpm. Mycelia were harvested via filtration and washed with distilled water. Then, 200 mg lyophilized mycelia were treated with 3 ml of 25% alcoholic potassium hydroxide (methanol to ethanol, 3:2), vortexed for 1 min, and incubated for 1 h at 85°C. After the samples cooled to room temperature, they were combined with 3 ml of pentane and 1 ml of distilled water and then vortexed for 3 min. After the samples stood still for 10 min, the upper hexane layer was transferred to a new tube. Then, the samples were evaporated in a fume hood and dissolved in 1 ml of methanol. After filtration through a pore-size 0.2 μm filter, the samples were analyzed by high-performance liquid chromatography (HPLC) (Agilent Technologies) with an AQ-C18 column (250 mm by 4.6 mm, 5 μm) for 15 min at 282 nm with a flow rate of 1 ml/min.
4
Sustained Anticancer Drug Release in Tumor Microenvironment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The normal microenvironment was pH = 7.4 PBS buffer. The tumor microenvironment was pH = 6.5 PBS buffer + H2O2 + GSH and the concentration of H2O2 and GSH was 1 mM. An 808 nm laser was added to the tumor microenvironment to verify efficacy. With PBS buffer as the release medium under the above three conditions, 1 mL NCTD Gel was removed to the dialysis bag with the interception molecular weight of 3000, and 1 mL PBS was added for dispersion. The dialysis bag was placed into 19 mL PBS buffer and released dynamically at 25 °C under 500 r·min−1. At 1, 2, 4, 6, 8, 10, 12, 24 h, 200 μL of dialysis solution was removed (and the same amount of releasing medium was added), respectively. The content of GA was determined by High performance Liquid Chromatography (Agilent, Agilent Technologies Co., LTD. USA), and the cumulative release of GA in NCTD Gel was calculated. The results were repeated three times.
Chromatographic conditions: AQ-C18 column (250 mm × 4.6 mm, 5 μm, Agilent) was used. The mobile phase consisted of (A) acetonitrile and (B) 0.1% (v/v) aqueous phosphoric acid solution. The gradient elution conditions were 0–5 min, 60% A. The flow rate was 1 mL/min and the injection volume was 5 μL. The detection wavelength was 250 nm and the column temperature was 25 °C.
Chromatographic conditions: AQ-C18 column (250 mm × 4.6 mm, 5 μm, Agilent) was used. The mobile phase consisted of (A) acetonitrile and (B) 0.1% (v/v) aqueous phosphoric acid solution. The gradient elution conditions were 0–5 min, 60% A. The flow rate was 1 mL/min and the injection volume was 5 μL. The detection wavelength was 250 nm and the column temperature was 25 °C.
5
Quantification of Liver, Kidney, and Fecal Urate and Purines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To detect the urate and purine contents of the liver, kidney and feces, 200 mg of sample was added to 5 mL trifluoroacetic acid, 5 mL formic acid, and 10 mL H2O and hydrolyzed at 90 °C for 15 min. After hydrolysis, the hydrolysate was cooled quickly, and the solvent was dried using a vacuum centrifugal concentrator (Thermo Fisher Scientific, Waltham, MA, USA). The sample was redissolved in the mobile phase and filtered through a 0.45 μm filter disk. HPLC analysis was performed using an Agilent 1200 series HPLC System. Sample solution (20 μL) was injected onto an Aq-C18 column (4.6 mm × 250 mm, 5 μm) (Agilent, Santa Clara, CA, USA). Elution was carried out using 0.007 mol/L KH2PO4-H3PO4. The column was maintained at room temperature. The flow rate was 1 mL/min, and detection was performed at 254 nm.
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!