| Content Uniformity HPLC Exercise February 2005 Instrumental Analysis |
| Tablet Preparation Procedure: 1. Place about 50 mL of the special distilled water into each of ten 100 mL volumetric flasks. NOTE: The special water has been spiked with an internal standard. 2. Drop one aspirin tablet into each flask. 3. Sonicate, swirl and shake until the tablet is completely dissociated. 4. Fill the flask to the mark with the special distilled water. Shake thoroughly. 5. Centrifuge an aliquot of about 10 ml. Withdraw about 2 mL of the supernatant liquid and place it into a small vial. Do not allow any solids to contaminate the vial. Standard Preparation Procedure: 1. Weigh out 300 milligrams of aspirin standard into a 100 mL volumetric flask. 2. To create a check standard, weigh another 300 milligrams of aspirin standard into second flask. 3. Label the flasks as standard and check standard respectively. 4. Dilute each flask to the mark with the special distilled water. Sonication will be required to bring the standard into solution. HPLC Analysis Use 10 uL injections of each solution. Use a C-18 column and a flow rate between1 and 2 ml/minute. The mobile phase is 50:50 methanol, water with 1% acetic acid. You will need to submit: Standard, check standard, each of the 10 samples from the volumetric flasks. Report: Your report must include the following: Manufacturer and lot number of the standard (aspirin) and the internal standard. Internal standard. Identification of, and the date of last calibration of the balance that was used to weigh out the standard and check standard. Identity of the HPLC system or systems used. Include the mobile phase, column type, last system calibration date and the date when the next calibration is due. Calculate the concentration of the check standard. It must be within 90% of the actual value. Use the procedures outlined below and treat the check standard as if it were an unknown. Once you have determined the check standard concentration, compare it to the known concentration. You will not use the check standard to quantitate the individual tablet results. Calculate the amount of aspirin in each of the volumetric flasks using the following procedure: Step one: Obtain the Factor (F) by injecting a known standard: Area Aspirin std peak Area of the internal standard peak --------------------------- = (F) ---------------------------------------- Concentration Aspirin Concentration internal standard Step two: Use the factor to calculate the amount of aspirin in the volumetric flask Area Aspirin peak Area of the internal standard peak --------------------------- = (F) ---------------------------------------- Concentration Aspirin Concentration internal standard Step three: Calculate the percent label strength of each tablet (%LS) %LS = (100) (conc. aspirin * 100 / stated label strength) Step 4: Report the %LS for each of the ten tablets, the average %LS and the standard deviation of all ten values. |
| Excerpts from an FDA Warning Letter about Content
uniformity: October 23, 2002 WARNING LETTER CERTIFIED MAIL RETURN RECEIPT REQUESTED Mr. Peter R. Dolan Bristol-Myers Squibb Manufacturing Company Worldwide and North American Medicines Chairman & Chief Executive Officer 345 Park Avenue, 44-61 New York, New York 10154-0037 Dear Mr. Dolan During an inspection of your manufacturing facility located in Mayaguez, P.R.; conducted on July 03, 2002 to August 01, 2002, our investigator documented deviations from the current Good Manufacturing Practice (cGMP) Regulations (Title 21, Code of Federal Regulations, Parts 210 & 2 11). These deviations cause your drug products, Cholestyramine Powder for Oral Suspension (Regular) ["Questran Powder"], Cholestyramine Powder for Oral Suspension (Generic Version), and Estrace tablets, to be adulterated within the meaning of Section 501(a) (2) (B) of the Federal Food, Drug, and Cosmetic Act. The cGMP violations documented during the most recent inspection include: 1. Failure to adequately validate the manufacturing processes for your drug product, Cholestyramine Powder for Oral Suspension (Regular), to assure uniformity and homogeneity in that you did not assess the adequacy of the mixing and filling processes and establish accurate mixing time limits. [21 CFR 211.110(a) (3); 21 CFR 211.111] a. No content uniformity samples are collected after the product is mixed and prior to being transferred to the holding tanks used during the filling operation, or throughout the filling operation. Thus, there is no assessment that the mixing process produces a homogeneous product and that the filling operation does not cause segregation of powder product. (483 items #5b, c) b. The manufacturing process validation for Questran Powder does not address the capability of the process to produce a homogeneous drug product in that the mixing time of the powder product is not established. Your manufacturing process controls are inadequate in that if, after the product is mixed and sampled during three different intervals, the results of any of these samples are out of specification for the assay test, your manufacturing instructions allow you to continue mixing (up to seven more minutes) until the results are within specifications. (483 item #5a) 2. Failure to have adequate laboratory controls in that once stability samples are found out of specifications and confirmed by re-test, you continue testing additional samples until passing results are obtained to conclude that the lot is within specification. [21 CFR 211.160; 21 CFR 211.166] We evaluated your written response, dated August 16, 2002, to the FDA form 483, …We conclude that you- have not satisfactorily addressed all of the noted deficiencies. Regarding your response to 483 item #4, it is unacceptable to release lots reporting initial out of specification results by testing additional samples, followed by the averaging of all results, i.e., those outside specification and others within specification. It is also unacceptable to apply the acceptance criteria of content uniformity in evaluating assay results to justify your practice of retesting and averaging results. We are requesting that you review all your initial out of specification results that were retested and averaged to obtain a final passing result and provide us, in your response to this warning letter, with a detailed description of the corrective action taken for the batches that were affected by your practice. We also expect to receive your written commitment to stop your retest and averaging practice… You should take prompt action to correct these deviations. Failure to promptly correct these deviations may result in regulatory action without further notice. These include seizure and/or injunction.... |