AEROSET® & ARCHITECT® cSystem™ MULTIGENT® Iron REF 6K95-50

Agenda Clinical Utility Methodology Sample Handling Reagent Handling Calibration Quality Control Reportable Range Interference Precision Method Comparison CAP Performance Troubleshooting tips Questions and Answers

MULTIGENT ® Iron Liquid Abbott will not market controls for U.S. use (Ranges for appropriate Bio-Rad controls have been established) Complaint handling: Abbott has responsibility for initial complaint registration. Sentinel (the vendor) will investigate level II complaints.

MULTIGENT ® Iron Liquid Assay disk ARCHITECT® MULTIGENT Assay Disk Version 2.00 8K50-02 Conventional/Alternate Units 8K51-02 SI/Alternate Units No AEROSET® Disk will be produced

Clinical Utility of Iron Iron exists in biological fluids as a component of hemoglobin and myoglobin and is bound in plasma to transferrin, which acts as a carrier protein. Increased iron concentrations occur in hemochromatosis and liver damage. Decreased iron levels can be caused by anemia due to malabsorption as a consequence of gastrointestinal diseases, or by blood loss as a result of the body’s iron status. The measurement of transferrin and ferritin can provide more detailed information.

MULTIGENT ® Iron Methodology Methodology: Ferene Chemical Reaction: Fe - transferrin pH 4.8 Fe+3 + transferrin Fe+3 Fe+2 Fe+2 + FERENE Fe – FERENE complex Hydroxylamine Hydrochloride

MULTIGENT ® Iron Methodology Description of Chemical Reaction: In a pH of 4.8 buffer system, iron is released from transferrin to which it is bound, and then quantitatively reduced to a ferrous state. The iron forms with Ferene-S* a stable colored complex of which the color intensity is proportional to the amount of iron in the sample. Particular reaction conditions and a specific masking agent almost entirely eliminate the interference from copper. * Ferene–S = 3-(2-pyridyl)-5,6-bis-[2-(5-furylsulfonic acid)]-1,2,4-triazine

MULTIGENT ® Iron Sample Handling Specimen Requirements: Serum Plastic tubes With or without gel barrier Plasma Sodium or Lithium Heparin Use nonhemolyzed serum collected by standard venipuncture techniques.

MULTIGENT ® Iron Sample Handling Sample Tubes tested and found acceptable: Vendor Ref Tube BD Europe 369032 Serum tube 367957 Serum with Gel separator 367869 Na - heparin 368884 Li-heparin (17 IU/mL) 367374 Li -heparin with PST II gel barrier

MULTIGENT ® Iron Sample Handling Specimen storage: Stored specimens must be inspected for particulates. If present, mix and centrifuge the specimen to remove particulates prior to testing. Sample volume = 20 μg/dL The system performs a 1:6.55 dilution of the specimen and automatically corrects the concentration by multiplying the result by the appropriate dilution factor. Maximum storage Temperature Serum/Plasma 20 to 25°C 7 days 2 to 8°C 3 weeks -20°C 1 year

MULTIGENT ® Iron Reagent Handling Reagent kit Reagent is liquid, ready-to-use, two reagent kit. Storage: 2 to 8°C. Store reagent in the box. Unopened kit is stable to the expiration date. Remove any air bubbles with an applicator stick. Reagent onboard stability is 30 days. List Number Kit Configuration REF 6K95-50 R1 3 x 90 mL R2 3 x 11 mL CAL 1 x 10 mL 904 tests/kit

MULTIGENT ® Iron Calibration Iron calibrator is liquid ready-to-use calibrator supplied in the reagent kit. Opened calibrator is stable for 120 days at 2 to 8°C if contamination is avoided and vials are recapped immediately after use. Calibration curve is stable for 14 days Calibrator value is verified using a NIST (National Institute of Standards and Technology), SRM 3126a.

MULTIGENT ® Iron Calibration Curve - ARCHITECT ®

MULTIGENT ® Iron Calibration Curve -AEROSET ®

Quality Control Abbott control product for Iron Liquid is not available, however BioRad controls are available for both the AEROSET® and ARCHITECT® customers BioRad peer value information can be found on WWCS webpage www.Bio-rad.com 1-800-2-BIO-RAD Control Product Name Catalog Number Configuration Lyphochek Assayed Chemistry Control L-1: C-310-5 L-2: C-315-5 12 x 5 mL Liquid Assayed Multiqual Chemistry Control L-1: 694 L-2: 695 L-3: 696 12 x 3 mL

MULTIGENT ® Iron Reportable Range Serum and Plasma Reportable Range = 5 to 1,000 μg/dL* (0.9 to 179.0 μmol/L) Limit of Detection (LOD)= 5 μg/dL* (0.9 μmol/L) LOD calculated on 10 replicates of normal saline reported as “mean zero value + 3 SD” *Note: SI units are μmol/L To convert results from μg/dL to µmol/L, multiply by 0.179.

MULTIGENT ® Iron Interference Interfering substances: MULTIGENT Iron Liquid assay is not affected (less than 10% difference) by the presence of the following interferents up to the concentrations indicated below: Interferring Substance Interferent Concentration Bilirubin 15 mg/dL (256.5 μmol/L) Hemoglobin 0.5 g/dL (5.0 g/L) Human triglyceride 1,000 mg/dL (11.3 mmol/L)

MULTIGENT ® Iron Precision Representative data from a 10 day study with two runs and two replicates of each control per day are summarized below. Control Level 1 Level 2 N Mean (μg/dL) 40 112.4 188.4 Within Run SD %CV 0.92 0.82 1.28 0.68 Between Run SD 1.53 1.36 3.64 1.93 Total SD 2.34 2.08 3.78 2.01

MULTIGENT ® Iron Method Comparison Serum results from the MULTIGENT Iron Liquid assay on an ARCHITECT c System and the AEROSET System were compared with results from the MULTIGENT Iron Liquid assay on a commercially available analyzer. c System were compared with the results on the AEROSET system. ARCHITECT vs. Comparative Method AEROSET vs. Comparative Method ARCHITECT vs. AEROSET N 58 60 40 Y – Intercept -0.40 2.89 1.65 Correlation Coefficient 0.9998 0.9996 0.9971 Slope 1.002 0.9584 0.9938 Range (μg/dL) 16 to 137 14 to 247 12 to 196

MULTIGENT ® Iron method comparison internal study An internal study was performed to determine the potential bias customers may observe when converting from the Abbott Iron assay (LN 7D68) to the MULTIGENT Iron assay. The study data and conclusions follow. MULTIGENT Iron Serum vs. Abbott Iron Serum Samples

MULTIGENT ® Iron method comparison internal study MULTIGENT Iron Plasma vs. Abbott Serum Samples Conclusion: Customers that convert to MULTIGENT Iron liquid from Abbott Iron may observe an average of 8.7% higher results when using serum samples.

MULTIGENT ® Iron CAP Performance CAP Proficiency Testing (PT) for MULTIGENT Iron assay was performed in-house on the Architect c8000 analyzer using CAP PT materials : CHM-01, CHM-02, CHM-03, CHM-04 and CHM-05. 2008 CZ-A CAP Survey Results (Information Only) MULTIGENT All Method Mean CHM-01 213.0 215.3 CHM-02 213.0 231.8 CHM-03 178.0 182.8 CHM-04 149.0 150.9 CHM-05 119.0 119.0 NOTE: results units = μg/dl

MULTIGENT ® Iron CAP Performance CAP Proficiency Testing (PT) for MULTIGENT Iron assay was performed in-house on the Architect c8000 analyzer using CAP PT materials : CHM-06, CHM-07, CHM-08, CHM-09 and CHM-10. 2008 CZ-B CAP Survey Results (Information Only) MULTIGENT All Method Mean CHM-06 177.0 176.3 CHM-07 73.0 70.9 CHM-08 148.0 146.4 CHM-09 88.0 86.1 CHM-10 221.0 221.7 NOTE: results units = μg/dl

Troubleshooting Tips AEROSET® system: Iron must be configured on a separate line from Creatinine. If running Microalbumin, you must configure Microalbumin assay parameters with the following SmartWashes: Component Reagent/Assay Wash Volume (μl) Reagent probe FEP0S61 AlkW 345 FEP0S62

Questions and Answers Why am I getting a No Assay reagent status? (ARCHITECT) Answer: The reagent is not used by any assay installed on the system. Ensure the reagent parameters are entered correctly. Enter the reagent name exactly as displayed in the reagent application sheet FEP0S. The reagent name contains a zero (0) not an alpha character.

Questions and Answers Package insert sample requirements state heparin salt is acceptable. Does this mean that all heparin is OK? Answer: Yes, all types of heparin (ammonium, lithium, and sodium) are acceptable.

Does copper interfere with the MULTIGENT Iron Liquid assay? Questions and Answers Does copper interfere with the MULTIGENT Iron Liquid assay? Answer: Particular reaction conditions and a specific masking agent almost entirely eliminate the interference from copper. In the MULTIGENT Iron package insert, the specific masking agent is identified as thiourea. Cited article statement: “The effects of copper interference can be almost entirely eliminated by the addition of thiourea, which preferentially binds the copper in a copper (1) complex. Reference: Gaudin, J. The determination of serum iron and total iron binding capacity. Diagnostic Chemicals LTD, Charlottetown, P.E.I., Canada. End of Document