Document oe8e4B4DdQbV2Vad18jb4v8N8

RM* - 3 * 6 0 442 DuPont Haskell Laboratory for Health and Environmental Sciences AIU ltf_3* TO: FROM: WMR FILE POCKET H-25425 1 J. C. Maslanka November 4, 2002 cc: M R jjp B J t/ Quality Assurance TEST SUBSTANCE STABILITY O F H-25425 Medical Research Project Number: Haskell Sample Number: Haskell Reference Standard Number: Analytical Test Code: Analytical Report Number: Notebook References: 25425 25425 HA-2002-094 Attached is the analytical report for the study identified above. This analysis was performed to satisfy protocol requirements for test substance stability but may also be used for other purposes. TEST SUBSTANCE STABILITY OF H-25425 M edical Research Project Number: Haskell Sample Number: Analytical Report Number: 25425 HA-2002-094 when the sample was submitted. ACKOWLEGEMENTS Sample preparation by Sheila A. Riley (Chemistry Associate). Sampie analysis by Bogdan Szostek (Senior Research Chemist). SIGNATURE Report by: Date Issued: Analytical Staff Chemist SL O O S-J HA-2002-094 page 2 of 2 METHODS Analysis for the percent o f a.i. in a sample of H-25425 was by high-performance liquid chromatography (HPLC) with a Gel Permeation Column (GPC) and refractive index detector according to the following method. Sonication was used to completely dissolve the test substance and the analytical standard stock solution. SAMPLE PREPARATION & ANALYSIS Aliquots (0.0413, 0.0407 and 0.0412 grams) o f H-25425 were dissolved in tetrahydrofuran (10 mL) to give nominal concentrations o f 4130,4070, and 4120 ppm, respectively. CHROMATOGRAPHIC CONDITIONS Instrument: Column: Mobile Phase Flow Rate: Injection Volume: Column Temperature: Detection: Hewlett-Packard Model ilOO HPLC Styrage.I HR 4; 7.8 m m x 300 mm (GPC column) 100% tetrahydrofuran (THF) 1.0 mL/min 50 pi 35C Refractive Index CALIBRATION & QUANTITATION A separate sample of H-25425 test substance was designated as the analytical reference standard (93.17% pure). A stock solution of H-25425 in tetrahydrofuran was used to make calibration solutions that bracketed the test solutions. Peak heights from replicate HPLC/GPC/RI analysis of each calibration solution were used to construct a calibration curve by least-squares regression. Measured concentrations for each test solution were determined by applying respective peak heights to the calibration curve. RESULTS H-25425 eluted from the HPLC column as a resolved p eak w ith a retention time o f about 8.582 0.01 minutes. The sponsor reported a purity o f l f B H ^ h e n the sample was submitted. T he percent of a.i. in the H-25425 sam ple analyzed O ctober ppm H-25425 Percent Aliquot Nominal Measured Nominal 1 4130 2 4070 3 4120 A verage % Nominal S tandard Deviation Coefficient of Variation 3730 3720 3790 90.3 91.4 92.0 91.2 0 .8 5 1%