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RECEIVED for review August 19, 1985. Accepted October 30, lg85*This work was supported by a grant from the National Science Foundation (Grant No. CHE-8312494).
Subambient Temperature High-Performance Liquid Chromatographic Determination of the Enantiomers of (f)-((6,7-Dichloro-2,3-dihydro-2-methyl- I-oxo-2-phenyl4 Hinden-5-yl)oxy)acetic Acid David J. Mazzo,* C a r l J. Lindemann, a n d Gerald S. B r e n n e r Department of Pharmaceutical Research and Development, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486
The enantiomers comprising the antihypertensive compound MK-286 were determined by subambient temperature hlghperformance liquid chromatography. A Pirkie covalent Lphenylglycine chirai stationary phase was used in conjunction with a predominantly (90% ) organic buffered ternary reversed-phase mobile phase to effect the separation. Subambient column temperatures to an optimum of 0 O C improved enantiomer resolutlon and provided reproducible chromatography. Under the assay conditions the separation was accompllshed in less than 15 mln. Both enantiomers were detectable to the low pg/mL concentratlon range with an assay accuracy within f2% and an assay precision (relative standard deviation) of 0.999. The subambient temperature reversed-phase HPLC determination of the enantiomers comprising MK-286 using a chiral stationary phase has become a routine assay in our laboratories for the testing of formulated product. This method has been shown to give statistically equivalent results
m g of MK-286 normalized"
m g of (+) enantiomer normalized"
m g of (-) enantiomer normalized"
0.945 0.941 0.934 0.953 0.957 0.968 0.972 0.968 0.975 0.981
0.973 0.935 0.928 0.943 0.950 0.961 0.969 0.960 0.967 0.975
1.00 1.00 0.994 1.01 1.02 1.03 1.04 1.03 1.03 1.05
0.959 1.50 1.57
0.952 1.54 1.61
1.02 1.78 1.75
'Indicates t h a t t h e assay value reported has been normalized t o t h e tablet weight.
for total drug concentration in tablets when compared to a conventional nonchiral specific HPLC method and, as such, has been used to monitor the optical stability (as the ratio of enantiomers) of MK-286 in aged tablets. The method has been reproducible and has prompted further investigation into the effects of low temperatures on chiral and nonchiral HPLC separations. Registry No. (f)-MK-286,57296-63-6; MK-199, 56049-89-9; MK-197, 57297-16-2.
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RECEIVED for review August 12, 1985. Accepted October 21, 1985.
