Certificate of Analysis

Transcription

1 Certificate of Analysis ISO GUIDE 34 ANAB Cert# AR-1470 ISO/IEC ANAB Cert# AT-1467 GENTAMICIN SULFATE CERTIFIED REFERENCE MATERIAL CERTIFIED PURITY: 637µg/mg, U crm = ±52.0µg/mg, k = 2 (Antibiotic Assay/dried basis) 567IU/mg, U crm = ±37.6IU/mg, k = 2 (Antibiotic Assay/as is basis) NOMINAL PACKAGE SIZE: 1g CATALOG #: PHR1077 LOT #: P CERTIFICATE VERSION: ISSUE DATE: 28 February 2017 Note: Certificates may be updated due to Pharmacopeial Lot changes or the availability of new data. Check our website at: for the most current version. CRM EXPIRATION: 31 December 2018 (Proper Storage and Handling Required). RECEIPT DATE: Note: this space is provided for convenience only and its use is not required. STORAGE: Store in a Freezer/Protect from Light, keep container tightly closed. Attachment of a 20 mm aluminum crimp seal recommended for unused portions. CHEMICAL FORMULA: NA MW: NA PHYSICAL DESCRIPTION: White powder in amber vial CAS #: HAZARDS: Read Safety Data Sheet before using. All chemical reference materials should be considered potentially hazardous and should be used only by qualified laboratory personnel. Page 1 of 7

2 INSTRUCTIONS FOR USE: For USP and BP applications, dry portion under vacuum at 110ºC for 3 hours prior to use. For EP applications, do not dry, use on the as is basis. The internal pressure of the container may be slightly different from the atmospheric pressure at the user s location. Open slowly and carefully to avoid dispersion of the material. This material is intended for R&D use only. Not for drug, household or other uses. TRACEABILITY ASSAY Comparative assay demonstrates direct traceability to Pharmacopeial Standards Specification: NLT 590µg/mg (USP) METHOD: ANTIBIOTICS-MICROBIAL ASSAY (ref.: USP General Chapter <81>) Test Organism: Staphylococcus epidermidis (ATCC 12228, Lot: ) Base and Seed Layer: Difco Antibiotic Medium 11 Agar (Lot: ) Incubation: 24 hours at 37ºC Plate Reader: Trinity V3 (Giles Scientific) ASSAY vs. USP REFERENCE STANDARD (dried basis) ASSAY VALUE U crm k value vs. USP LOT 637µg/mg ±52.0µg/mg 2 M1J001 ASSAY vs. EP CRS (as is basis) ASSAY VALUE U crm k value vs. EP BATCH 567IU/mg ±37.6IU/mg ASSAY vs. BP CRS (dried basis) ASSAY VALUE U crm k value vs. BP BATCH 628.7µg/mg ±51.3IU/mg Labeled Content = 697µg/mg Labeled Content = 17352IU/vial Labeled Content = None Assigned Content = 661 µg/mg* *The assigned content of the BP was determined by comparison against the USP Reference Standard Page 2 of 7

3 Representative Zone Measurement Picture Page 3 of 7

4 CHROMATOGRAPHIC CONTENT METHOD: HPLC (ref.: Gentamicin Sulfate, USP33) Column: Wakosil 5C18RS, 4.6 x 150mm, 5µm Mobile Phase: 5g/L Sodium-heptanesulfonate in Methanol, Water, Acetic Acid (70:25:5) Flow Rate: 1.5mL/min Column Temperature: 30º Injection: 20µL Detector: 330nm Derivatizing solution: 10mg/mL o-phthalaldehyde in Methanol, 0.4M Boric Acid (5:95) Calculated Content: Gentamicin 1: 30.1% Gentamicin 1a: 28.3% Sum of Gentamicin 2: 41.6% Representative Chromatogram from Lot: P Content Analysis METHOD: GC (ref.: Gentamicin Sulfate, USP33) Column: DB-Wax, 30m x 0.320mm x 0.50µm Carrier gas: N 2 Flow: 1.2mL/min Split Ratio: Splitless Injection/Temperature: 1µL/240 C Temperature Program: 130 C for 7min Detector/Temperature: FID/240 C Internal Standard: n-propyl Alcohol Solvents Detected: None Page 4 of 7

5 LOSS ON DRYING/VOLATILES Method: Under vacuum 100ºC Mean of three measurements, Loss = 9.4% RESIDUE ANALYSIS Method: Sulfated Ash Sample Size: ~1g Mean of three measurements, Residue = 0.1% IDENTIFICATION TESTS INFRARED SPECTROPHOTOMETRY (Comparative identification analysis demonstrates direct traceability to Pharmacopeial standards) Page 5 of 7

6 1 H NMR (Data provided by an external laboratory; not in scope of accreditation) Consistent with structure OPTICAL ROTATION Specification: +107º to +121º (USP) Perkin Elmer Polarimeter 343 Wavelength: 589nm Concentration: ~1g/100mL Cell Path: 100mm Mean of three Measurements = Page 6 of 7

7 HOMOGENEITY ASSESSMENT Homogeneity was assessed in accordance with ISO Guide 35. Completed units were sampled using a random stratified sampling protocol. The results of chemical analysis were then compared by Single Factor Analysis of Variance (ANOVA). The uncertainty due to homogeneity was derived from the ANOVA. Heterogeneity was not detected under the conditions of the ANOVA. Analytical Method: Antibiotic Assay Sample size: ~40mg UNCERTAINTY STATEMENT Uncertainty values in this document are expressed as Expanded Uncertainty (U crm ) corresponding to the 95% confidence interval. U crm is derived from the combined standard uncertainty multiplied by the coverage factor k, which is obtained from a t- distribution and degrees of freedom. The components of combined standard uncertainty include the uncertainties due to characterization, homogeneity, long term stability, and short term stability (transport). The components due to stability are generally considered to be negligible unless otherwise indicated by stability studies. STABILITY ASSESSMENT Significance of the stability assessment will be demonstrated if the analytical result of the study and the range of values represented by the Expanded Uncertainty do not overlap the result of the original assay and the range of its values represented by the Expanded Uncertainty. The method employed will usually be the same method used to characterize the assay value in the initial evaluation. Long Term Stability Evaluation - An assessment, or re-test, versus a Compendial Reference Standard may be scheduled, within the 3 year anniversary date of a release of a Secondary Standard. The re-test interval will be determined on a case-by-case basis. Short Term Stability Study - It is useful to assess stability under reasonably anticipated, short term transport conditions by simulating exposure of the product to humidity and temperature stress. This type of study is conducted under controlled conditions of elevated temperature and humidity. QC Supervisor QA Supervisor APPENDIX Original Release Date: 30 June 2014 Requalification Test Date: 27 November 2014 Requalification Test Date: 28 February 2017 Manufactured and certified by Sigma-Aldrich RTC, Inc Soldier Springs Rd, Laramie WY, USA (Phone): (Fax): RTCTechGroup@sial.com Page 7 of 7