HPLC Analysis of Bendamustine and Related Impurities

  Application Notes: Bendamustine is a hydrophobic and basic drug, classified as nitrogen mustard. Two approaches were used to separate bendamustine from related impurities. In one approach, a Primesep 200 HPLC column was used in reversed-phase cation-exchange mode. Primesep D column was also used in reversed-phase cation-exclusion mode. Both methods are alternative to each other. Both can be used to quantify bendamustine in various sample matrices (drug composition, blood, plasma, urine, etc.). Sample preparation is required in the case of biofluids using the Primesep 200 column. Primesep D can be used for direct analysis of bendamustine in blood, plasma and urine. Proteins in the biofuids repel from stationary phase. Sulfuric acid in the mobile phase can be replaced with ammonium formate or acetate for LC/MS and with TFA for ELSD/CAD. Application Columns: Primesep 200, Primesep D Application compounds: bendamustine and related impurities Detection technique: UV, LC/MS, ELSD/CAD

Condition 1

Column Primesep 200, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN - 30%
Buffer H2SO4 - 0.1%
Flow Rate 0.6 ml/min
Detection UV, 215 nm

Condition 2

Column Primesep D, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN - 15%
Buffer H2SO4 - 0.1%
Flow Rate 1.0 ml/min
Detection UV, 230 nm


Class of Compounds Drug, Chemotherapy, Hydrophilic, Ionizable
Analyzing Compounds Bendamustine
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Application Analytes:
HPLC Analysis of Bendamustine and Related Impurities

Application Detection:
UV Detection
ELSD/MS Detection