5-Bromo-2-Chloropyrimidine CAS 32779-36-5 Macitentan Intermediate Purity ≥99.5% (HPLC)

1 Method of Analysis
High performance liquid chromatography method for the purity detection of 5-Bromo-2-Chloropyrimidine.
2 Sample Information
Sample name: 5-Bromo-2-Chloropyrimidine
CAS: 32779-36-5
Properties: White to off-white powder
3 Sample purity analysis
3.1 Instruments and equipment
Shimadzu LC-20AT liquid chromatograph
Purified water
Analytical balance
Chromatographic column YSC-PACK ODS-A/S-5um/12nm(150mm*6.0mm)
3.2 Reagents and samples
Purified water
Acetonitrile (ACN, chromatographically pure)
Trifluoroacetic acid (TFA, chromatographically pure)
QN-A12 Sample
3.3 Solvent Preparation
3.3.1 Solvent
Acetonitrile: Water = 50:50
3.3.2 Probe rinse solution
Acetonitrile
3.3.3 Mobile phase
Phase A: Water (0.1% TFA, v/v)
For example: precisely remove 0.5 mL of TFA and add it to 1000 mL of pure water, mix it and filter it and degass it ultrasonic
B phase: acetonitrile
Example: acetonitrile filtration and ultrasonic degassing.
3.3.4 Sample Solution (0.4mg /mL)
Make two copies in parallel. For example, a 20 mg sample of 5-Bromo-2-Chloropyrimidine was accurately weighed in a 50 mL volumetric bottle, dissolved with solvent and fixed to the scale, shaken and then used for injection, labeled as sample solution #1. Another sample solution, labeled #2 sample solution, was prepared in the same way.
3.4 Test Procedures
3.4.1 Chromatographic conditions
Chromatographic column YSC-PACK ODS-A/S-5um/12nm(150mm*6.0mm)
Wavelength UV 220 nm
Column temperature 35 ℃
Flow rate: 1.0 mL/min
Injection volume: 5 µL
Mobile phase
A: 0.1% TFA in Water (v/v)
B: ACN
Equal degree program A: 50%
B:50%
Data collection time: 30 minutes
Solvent: Acetonitrile: Water = 50:50
Probe rinse solution: Acetonitrile
Note: The above conditions are based on Shimadzu LC-20AT instrument, some conditions such as rebalancing time can be adjusted according to the actual situation.
3.4.2 Sampling sequence
Please refer to the following table for sampling sequence.
Number Injection solution Number of injection needles
1 Blank (solvent) ≥1
2 #1 1
3 #2 1
3.4.3 Reference retention time
Compound ID Retention Time (min)
5-Bromo-2-Chloropyrimidine 10.8
3.5 Calculation and report
The purity of the sample is calculated according to the peak area normalization method, if the results of both samples meet the quality specification and the RSD between the two results is not greater than 2.0%, the reported average is retained to one decimal place.