Thomas L. Barry

Isolation and identification of process impurities in trimethoprim drug substance by high-performance liquid chromatography, atmospheric pressure chemical ionization liquid chromatography/mass spectrometry and nuclear magnetic resonance spectroscopy.

Twenty-two lots of recently synthesized trimethoprim drug substance, from five different manufacturers, in three different countries of origin, China, Israel and the United States, were investigated for the presence of impurities. A liquid chromatographic system, using gradient elution, and a mobile phase consisting of 0.25% TEA/0.1% formic acid (pH 5.8)--acetonitrile, was used to separate and detect two significant, recurring impurities in trimethoprim drug substance. The two impurities were isolated by preparative liquid chromatography and identified, using a combination of liquid chromatography/mass spectroscopy and nuclear magnetic resonance, as 2,4-diamino-5-(4-ethoxy-3,5-dimethoxybenzyl) pyrimidine and 2,4-diamino-5-(3-bromo-4,5-dimethoxybenzyl) pyrimidine. These impurities were not detected by the compendial method and were present at significant levels in 17 of the lots tested. Total impurity concentrations were in the range of 0.1-2.1%.

Isolation of a 2:1 hydrochlorothiazide–formaldehyde adduct impurity in hydrochlorothiazide drug substance by preparative chromatography and characterization by electrospray ionization LC–MS

Hydrochlorothiazide drug substance (19 lots) from five different manufacturers and four different countries of origin (USA, Italy, Hungary, and Croatia) were analyzed for the presence of impurities using a gradient elution chromatographic system, with acetonitrile-water as the mobile phase. Two known impurities of hydrochlorothiazide, 4-amino-6-chloro-1,3-benzenedisulfonamide and chlorothiazide, were separated, as well as a late-eluting, unknown, recurring impurity. The unknown impurity was isolated by preparative liquid chromatography followed by preparative thin-layer chromatography. It was characterized by electrospray ionization LC-MS as a 2:1 hydrochlorothiazide-formaldehyde adduct of the parent drug substance. The adduct is believed to form through the double condensation reaction of hydrochlorothiazide with excess formaldehyde during the parent compounds synthesis. The concentration of this impurity ranged from 0.02 to 1.1% (area%), and was above the 0.1% USP Other Impurities threshold in 16 of the 19 lots examined.

LC determination of impurities in methoxsalen drug substance: isolation and identification of isopimpinellin as a major impurity by atmospheric pressure chemical ionization LC/MS and NMR.

A gradient elution LC method was developed to separate methoxsalen from three of its known impurities: isopimpinellin, bergapten, and ammidin. The method employs a methanol-6%THF (aq) mobile phase, phenyl column, and detection at 254 nm. The gradient LC procedure was applied to seven lots of methoxsalen from five different manufacturers. Six of the seven lots tested contained isopimpinellin as the major impurity at a concentration range of 0.2-2.5%. Identification of the impurity as isopimpinellin was accomplished by a combination of analytical and preparative LC, atmospheric pressure chemical ionization liquid chromatography/mass spectrometry, and NMR.

Identification of pentachlorobenzonitrile residues in field crops.

We recently received several carrot extracts from another FDA district laboratory for identification of a material present in each of the extracts which had a GLC retention time inconsistent with that of any known pesticide. The material, which was eluted from Florisil with 15% ether in petroleum ether (AOAC, par. 29.015, 1975) exhibited both electron capture and microcoulometric responses (LASKI and LEONE, 1976). This paper reports structural elucidation, GLC characteristics and recovery data for the material.