Samples were prepared using our regular recordings solution without adding trifluoroacetic acid to maintain a neutral pH. We injected 50 nmol of 2-APB and 5 nmol of CBD, either by themselves or together, and the final volume injected into the HPLC column was 400 µL with the compound diluted in dH2O. We injected samples into a 5 µM Ultrasphere C18 column (Beckman Coulter, Brea, CA) on a 1525 Binary pump HPLC system (Waters Corporation, Milford, MA), and used a gradient of 0% to 100% acetonitrile over 40 min and monitored sample absorbance at 228 nm.
Ultrasphere c18 column
Manufactured by Beckman Coulter
Sourced in Canada
The Ultrasphere C18 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a silica-based stationary phase with octadecyl (C18) functional groups, which allows for the effective separation of both polar and non-polar analytes. The column's construction and packing ensure efficient and reproducible chromatographic performance.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using ultrasphere c18 column
1
Quantification of 2-APB and CBD
2
Synthesis and Purification of Fluorine-Containing Peptide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To the peptide 10 (253 mg, 0.22 mmol) in degassed PBS (150 mL) was added
a solution
of compound9 (196 mg, 0.27 mmol) in degassed EtOH (30
mL). The mixture was stirred at rt under argon and monitored by analytical
reversed-phase high performance liquid chromatography (RP-HPLC). The
mixture was quenched by 0.1% aqueous TFA and concentrated through
rotary evaporation. The residue was purified by preparative HPLC.
The proper fraction was collected and lyophilized to afford fluorine-containing
peptide as a white solid (316 mg, 76% yield). Mass (ESI) m/z 942.6 [M + 2H]2+. 19F NMR
(282 MHz, D2O) δ 70.50. For semipreparative HPLC,
a Beckman Ultrasphere C18 column (10 × 250 mm) and
a gradient elution profile were used with 0.5% phosphoric acid in
water (solvent A) and 0.5% phosphoric acid in CH3CN (solvent
B). The elution profile was isocratic at 5% solvent B for 5 min, then
a gradient to 80% solvent B over 45 min. The flow rate was 4 mL/min.
The major peak at about 27.0 min was collected. The purity of the
resulting compound was conducted by analytical HPLC.
a solution
of compound
mL). The mixture was stirred at rt under argon and monitored by analytical
reversed-phase high performance liquid chromatography (RP-HPLC). The
mixture was quenched by 0.1% aqueous TFA and concentrated through
rotary evaporation. The residue was purified by preparative HPLC.
The proper fraction was collected and lyophilized to afford fluorine-containing
peptide as a white solid (316 mg, 76% yield). Mass (ESI) m/z 942.6 [M + 2H]2+. 19F NMR
(282 MHz, D2O) δ 70.50. For semipreparative HPLC,
a Beckman Ultrasphere C18 column (10 × 250 mm) and
a gradient elution profile were used with 0.5% phosphoric acid in
water (solvent A) and 0.5% phosphoric acid in CH3CN (solvent
B). The elution profile was isocratic at 5% solvent B for 5 min, then
a gradient to 80% solvent B over 45 min. The flow rate was 4 mL/min.
The major peak at about 27.0 min was collected. The purity of the
resulting compound was conducted by analytical HPLC.
3
Reverse-Phase HPLC Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reaction progress was monitored by reverse-phase HPLC (Rp-HPLC) using a Beckman System Gold HPLC (Fullerton, CA) equipped with a 126 solvent module and 168 UV detector (λ = 254 nm) controlled by 32 Karat software and Beckman Ultrasphere C18 column (ODS, 4.6 × 250 mm, 5 μm), mobile phase: (A) 90% (0.1% TFA in water); (B) 10% (0.1% TFA in acetonitrile); 0–5 min, 10–90% B, 5–15 min, 90% B, 15–25 min. The flow rate was set to 1 mL/min. TLC was carried out on Merck silica gel 60 TLC plates F254 and visualized by UV at 254 nm. Column chromatography was performed using silica gel 60 (70–230 mesh). 1H and 13C were NMR recorded on a Bruker Avance 300 instrument with a deuterated solvent. Mass spectra were recorded with a Waters (Waltham, MA USA) LCT Premiere ESI TOF mass spectrometer. The instrument was operated in positive ion ESI mode at a mass resolution of 10,000.
4
Synthesis of Fluorinated DOTA-Peptide Conjugate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A DOTA-containing
peptide (75 mg) was dissolved in PBS, GdCl3·6H2O (5 equiv) was added, and the pH of the
solution was adjusted to 4–5. The mixture was heated at 80
°C, and the reaction was monitored by HPLC; typically the reaction
was completed in 4 h. The mixture was centrifuged and subject to semipreparative
HPLC. A Beckman Ultrasphere C18 column (10 × 250 mm)
and a gradient elution profile were used with 0.5% phosphoric acid
in water (solvent A) and 0.5% phosphoric acid in CH3CN
(solvent B). The gradient elution profile was from 5% solvent B to
80% solvent B in 50 min, then to 100% solvent over the next 5 min.
The flow rate was 4 mL/min. The major peak at 34.4 min was collected
and lyophilized. Analytical HPLC was used to confirm the purity (Condition:
4.6 × 150 mm Phenomenex C18 column, 1 mL/min, detection
wavelength at 214 nm, elution profile, a gradient from 5% solvent
B to 60% solvent B in 15 min, and to 100% solvent B over next 5 min).
The retention time of the fluorinated peptide was 13.7 min. Mass (ESI) m/z 1019.6 [M + 2H]2+.
peptide (75 mg) was dissolved in PBS, GdCl3·6H2O (5 equiv) was added, and the pH of the
solution was adjusted to 4–5. The mixture was heated at 80
°C, and the reaction was monitored by HPLC; typically the reaction
was completed in 4 h. The mixture was centrifuged and subject to semipreparative
HPLC. A Beckman Ultrasphere C18 column (10 × 250 mm)
and a gradient elution profile were used with 0.5% phosphoric acid
in water (solvent A) and 0.5% phosphoric acid in CH3CN
(solvent B). The gradient elution profile was from 5% solvent B to
80% solvent B in 50 min, then to 100% solvent over the next 5 min.
The flow rate was 4 mL/min. The major peak at 34.4 min was collected
and lyophilized. Analytical HPLC was used to confirm the purity (Condition:
4.6 × 150 mm Phenomenex C18 column, 1 mL/min, detection
wavelength at 214 nm, elution profile, a gradient from 5% solvent
B to 60% solvent B in 15 min, and to 100% solvent B over next 5 min).
The retention time of the fluorinated peptide was 13.7 min. Mass (ESI) m/z 1019.6 [M + 2H]2+.
5
Cy5.5-labeled Compound Synthesis
6
Cy5.5 Dye Conjugation to Compound 5
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!