A modified HPLC-based method to quantify bilirubin levels was used [25 (link)]. Plasma (200–300 µL) was diluted 1:1 with acetonitrile (Honeywell, Germany) and centrifuged at 20,000× g for 10 min at 4 °C, and 200 µL of clear supernatant was transferred to a glass vial and put into the HPLC autosampler together with the bilirubin standards (Sigma-Aldrich). Standards of 5 µM bilirubin in H2O/acetonitrile (1:1) were used with a total volume of 200 µL. The HPLC system comprised a control unit, two pumps, a mixer, detectors, a degasser, an autosampler (AS-2057 plus) from Jasco Series 2000 (Groß-Umstadt, Germany), and a UV–VIS detector (UV-2077 Plus). The column (C18-Nucleosil 100−3 (125 × 4)) was purchased from Macherey-Nagel, Düren, Germany. The high-pressure gradient was based on the mobile phases 90% acetonitrile and 5 mM citrate buffer, pH 2.2. The gradient conditions were as follows: minute 00:00, 50% A: 50% B; minute 10:00–18:00, 0% A: 100% B; minute 19:00–20:00, 50% A: 50% B. The flow was set to 1 mL/min, and bilirubin was detected by its absorption at 450 nm at a retention time of 15 min and 45 s.
Uv 2077 plus
Manufactured by Jasco
Sourced in Germany
The UV-2077 Plus is a high-performance UV-Visible spectrophotometer designed for accurate and reliable measurements in laboratory settings. It features a wide wavelength range, advanced optics, and precise data acquisition capabilities, making it suitable for a variety of analytical applications.
Automatically generated - may contain errors
3 protocols using uv 2077 plus
1
Quantification of Bilirubin Levels by HPLC
2
HPLC Analysis of Hydrochlorothiazide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC system consisted of a Jasco chromatograph equipped with a quaternary pump (PU-2089s Plus) and a Jasco Multiple Wavelength detector (Jasco UV-2077 Plus) set at 270 nm. Chromatographic separations were performed on a C18 Restek (15 cm x 4.5 mm x 5 µm) column thermostatized at 30 ºC. A Phenomenex Security Guard Fusion® RP (4 x 30 mm) guard column was also used. Chromatographic analyses were performed in the gradient mode, with the mobile phase consisting of water (pH = 3.6, adjusted with acetic acid):
Methanol HPLC grade (MeOH HPLC ) (80:20, v/v) at a flow rate of 1 mL/min from 0 to 5 minutes (min), which linearly changed to a 50:50 v/v mixture from 5 to 9 min and that was kept constant from 9 to 11 min. Finally, the solvent composition was linearly modified to its original rate (80:20, v/v) in 1 min. In addition to the time required for the injection cycle of the instrument, the injection time, and the needle wash cycle (3 min), the full equilibration time between runs was approximately 4 min to ensure the column re-equilibration prior to the following injection. 10 consecutive injections of HCT standards were performed to ensure complete system re-equilibration, and no significant changes were found in HCT retention times. The total run time was 12 min, with 15 µL injection volumes being applied.
Methanol HPLC grade (MeOH HPLC ) (80:20, v/v) at a flow rate of 1 mL/min from 0 to 5 minutes (min), which linearly changed to a 50:50 v/v mixture from 5 to 9 min and that was kept constant from 9 to 11 min. Finally, the solvent composition was linearly modified to its original rate (80:20, v/v) in 1 min. In addition to the time required for the injection cycle of the instrument, the injection time, and the needle wash cycle (3 min), the full equilibration time between runs was approximately 4 min to ensure the column re-equilibration prior to the following injection. 10 consecutive injections of HCT standards were performed to ensure complete system re-equilibration, and no significant changes were found in HCT retention times. The total run time was 12 min, with 15 µL injection volumes being applied.
3
HPLC Analysis of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HPLC analyses were done using a Jasco chromatograph equipped with a quaternary pump and a Jasco Multiple Wavelength detector (Jasco UV-2077 Plus) set at 270 nm. To perform chromatographic separations, a Phenomenex Synergy Fusion C18 analytical column (4.0 x 250 mm, 5 mm particle size) coupled to a Phenomenex Security Guard Fusion RP (4.0 x 30 mm) guard column was used. chromatographic analyses were carried out using mobile phase gradients at a flow rate of 1 mL/min as follows: 0-2.5 mins: a mixture of a pH 2.0 phosphate buffer (0.1 mM): methanol (53:47, v/v), with a linear change between 2.5-9.5 mins to a 30:70 v/v composition. The mobile phase composition was kept constant from 9.5-12.0 mins. To ensure the re-equilibration of the mobile phase composition and the column conditions, the solvent gradient was reverted to initial conditions for 4.0 mins before the next injection was performed. The total run time for each injection was 15 min, with a sample injection volume of 10 μL.
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!