Product no.: Lot no.: Description of CRM: Expiry date: Storage: Certificate version: Chemical formula: Molecular mass: CAS No.: EO FOLIC ACID weg PHR1035-16 LT Au LRAC7879 v Yelow Powder 31 October 2024 Room Temperature LRAC7879.1 te: crater ay bpd ae Hise as Cheeta tc sa oar soe Ce ca teen eed Gitte aaa 59-30-3 Certified Purity 4 associated uncertainty U, U=k-u (k=) (Mass Balante/ ren} Folic Acid Metrological traceability: Measurement method: Intended use: Minimum sample si2« Instructions for handling and correct use: Health and safety information: Accreditation: Certificate Issue date: atfas 15017034 AR-1470 96.3 % Uerm = + 0.8 %, k = 2.0 (anhydrous basis) 91.7 % Ucrm = + 0.8 %, k = 2.0 (as is basis) Traceable to the SI and higher order standards from NIST through an unbroken chain of comparisons, Additional traceability to Primary Standards is established through comparative assay determinations. See "Details on metrological traceability" on page 2. Where applicable, the certified value is based on a purity determination by mass balance, See "Certification process details” on page 3. Intended for RAD and Analytical Use only. Not for drug, household or other uses Bmg For USP applications, determine water content prior to use, and use on the anhydrous basis. For EP and BP applications, use on the as is. The internal pressure of the container may be slightly different from the atmospheric pressure ‘al Ue user” s location. Open slowly and carefully to avoid dispersion of the ‘material. Attachment of a 20 mm aluminum crimp seal recommended for unused portions, All chemical reference materials should be considered potentially hazardous and should be used only by qualified laboratory personnel. Please refer to the Safety Data Sheet for detailed information about the nature of any hazard and appropriate precautions to be taken. Sigma-Aldrich RTC Is accredited by the US accreditation, authority ANAB as a Fegistered reference material protlucer AR-1470 in accordance with ISO 17034. 02 February 2021 Gy a yp on bar [Andy Ommen; Quality Control} [Mark Pooler; Quality Assurance} Slams: Alsen RTC, 293% Soller Springs Rd. Laramie, WY 82070, USA, Tol. 1 307-742-5452; Fox 1 307-885 3-931 www sigmoalarich com ‘Sigma-Aldrich RTC Is 2 subsishary of Merck KGaA, Darmstadt, Germany, Ceniiieate Page 1 of S Certificate version 01etn tet ne ete ece tS At A PARE Packaging: 1g In amber vial Details on metrological This standard has been gravimetrically prepared using balances that have been traceability ally quai alibrated to 180 17028 requirements. All calibrations ut NIST traceable weights whieh are calibrated externally by a qualifie 702 accredited calibration laboratory to NIST standards, Qualification-of each balance Includes the assignment of a minimum weighing by a qualified and 1S0 +7025 ideration the balance and installed accredited calibration vendor taking inte cor environmental conditions to ensure compliance with USP tolerances of NMT 0.10% relative error. Fill volume ta predetermined specifications is Gravimetrically verfied throughout the dispensing process using qualified and calibrated balances. Further traceability to a corresponding Primary Standard may be achieved through a direct comparison assay. Where a Primary Stancard is available, the assay value will be included in the specified section of the COA Associated uncertainty: Uncertainty values in this document are expressed as Expanded Uncertainty (Ueu:) corresponding to the 95% confidence interval. Ucay is derived from the combined standard uncertainty multiplied by the coverage factor k, which is obtained from {@ 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. Assay: Comparative assay demonstrates direct traceability to Pharmacopelal Standards ASSAY Vs; USP REFERENCE STANDARD (anhydrous) z i Some Rosen Labeled Content = 0,989 ma/ma ASSAY vs. EP CRS (as is basis) ASSAY VALUE vs EP BATCH 91.8 % 2.0 ° Labeled Content = 90.7 % ASSAY Vsi BP CRS (a5 1s Basis) a 92.5% : ae " Labeled Content = 91.2 % Method: HPLC (ref.: Folic Acid, Current Compendial Monographs) Column: Ascentis Express C18, 25 cm x 4.6mm, Sjim particle size Mobile Phase: 2.0 g/L Potassium Phosphate Monobasic in 70.8% Water, 0.7% 3N Phosphoric Acid, 1.5% 0.5M Tetrabutylammenium Hydroxide in Methanol, 27% Methanol, pH: 5,0 Mobile Phase Ratio: 100 Flow Rate: 1.2 ml/min. Column Temperature: 25 °C Injection Volume: 10 pL. Detector: Wavelength: 280 nm Representative Chramatogram from Lot: LRAC7879 Analysis |sso te ME RE Se SEMA NGPA HAAR REALS = Certification process d ‘The certified purity Is determined by mass balance and calculated as 100 TCI “100 % Purity = (100 ~ ROI —LOD ~ H,0 ~ RS) + ( Tcl = Total Chromatographic Impurities LOD = Logs on Drying H30 = Water content determined by Karl Fischer analysis ROT = Residue on Tonition RS = Residual Solvents Methods for impurity determination may be added or deleted as required, The following techniques are applied: CHROMATOGRAPHIC IMPURITY ANALYSIS METHOD: HPLC (ref.: Folic Acid, Current Compendial Monographs) ‘See HPLC Assay Impurities Detected: Impurity 1: 0.374 % Impurity 2: 0.788 % Impurity 3: 0.316 % Impurity 4: 0.613 % Impurity 0.154 % Impurity 6: 1.310 % Impurity 1: 0.116 % Total Impurities: 3.671 % ‘Chromatogram from Lot: LRAC7879 Impurities Analysis Rep RESIDUAL SOLVENTS Method: GC-MS Headspace (ref.: Adapted from Residual Solvents USP <467>) Column: SPB-624, 30 m x 0.25 mm x 1.4 4m. Carrier gas: He Flow: 1.2 mL/min Split Ratio: 1:5 =fhfection/Temperature: 1 mL/220 °C ““ATemperature Program: 40 °C for 5 min, 8 °C/min to 200°C; hold § min a Solvents Detected: None WATER DETERMINATION Method: Karl Fischer (ref.: Current Compendial Monographs) Mean of three measurements, Water Content'= 4.78 % RESIDUE ANALYSIS Method: Sulfated Ash (ref.: Current Compendial Monographs) Sample Size: ~ 90 mg Mean of three measurements, Residue = 0.048 % CERTIFIED PURITY BY MASS BALANCE 96:3°% Ucrm = + 018146, k = 2.0 (anhydrous basis) 91.7 % Uerm = + 0.8%, k = 2.0 (as is basis) fate P Certificate version Yance 0 ee SREP ESN I LNT Homogeneity Homogeneity was assessed In accordance with ISO Guide 35. Completed units assessmen were sampled using a randam 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 rat detected under the conditions of the ANOVA. Analytical method: HPLC Sample size: 8 mg Significance of the stabllity 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 @ Compendia! 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 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. Stability assessment Identification Test: INFRARED SPECTROPHOTOMETRY (Comparative identification analysis demonstrates direct traceability to Pharmacopeial standards) Millipore = . a We Ae eT 3 3 %! ———— 5 ~~"2600 Waveuusnbers (eu-1) "3500 3000 3 minliporeSigma Lot: LRAC7879 vs, EP 9.0 / USP RUS6HO / BP 3282 Tarifas Page 3 oF S Tanicaye version OF“esr mteseceer re ne TnE ET nNSAaeaer e nse oHT ene NRPS ETH Indicative Values: MASS SPECTRUM Method: HR-QTOF; 4.0 kV €SI-; temperature: 325 °C x10 & [Foe Aca; 44s,1347:. Sean (et 2.170 min PHR1035, Folic Acid LRACISOG 3 2 25 $ e é 2 4 15 0? 1 8 Re os. e 5 2 Pleas 8 300 200 300 400 $00 600 700 000 000 1000 1100 1200 1300 1400 1500 1600 (Counts vs, Mase-te-Charge (mlz) ‘Theoretical value: 440.1319 m/z ‘The signal of the MS spectrum is consistent with the theoretical value and Its interpretation Is consistent with the structural formula, OPTICAL ROTATION ‘Specification: +18 to +22 ° (EP) Perkin Elmer Polarimeter 343 Wavelength: 589 nm Concentration: ~10 mg/mL in NaOH Cell Path: 100 mm, ‘Mean of three Measurements = 18,715 ° Certificate of analysis revision history: Certificate version Date Reason for version TRAC7879.1 02 February 2021 Original Release Disclaimer: The purchaser is required to determine the sultability of this product for any particular application, Sigma-Aldrich RIC makes no warranty of any kind, express or implied, other Urany its products meet all quality control standards set by Sigma-Aldrich RTC. We do not guarantee that the product can be used for any particular application. Al thar ragemarts ar te property ofthe respectve - ‘etaadinermatin on waderares ovate vn pbhaly aeeeatie etaurts, SaDst Mer KGAA, Davnsade, Germany arlor He hlotes. Al Rights Resend The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the US and Canada. Tonifieate Page 5 0 Tenifcate version OF