1. AEROSET® & ARCHITECT® cSystem™
MULTIGENT® Lithium REF 8L25
Target Launch: EU December 2007
US January 2008

2. Agenda Launch Strategy Clinical Utility Methodology Sample Handling
Reagent Handling
Calibration
Quality Control
Reportable Range
Interference
Precision
Method Comparison
CAP Performance
Reaction Graphs
Troubleshooting tips
Questions and Answers

3. MULTIGENT® Lithium Launch Strategy
There will be separate list numbers assigned to the reagent and calibrator products for E.U. and U.S. use.
U.S. list numbers will be distributed from Chicago for use in Canada, Latin America and the U.S.
E.U. list numbers will be distributed through Delkenheim to remaining countries.
Controls
Clin Chem Control 1 ( REF 6K3020) and Clin Chem Control 2 ( REF 6K3021) are available for E.U. use.
Abbott will not market controls for U.S. use (Ranges for appropriate Bio-Rad controls have been established)
There are no differences in the application or formulation between the U.S. and E.U. products.
Complaint handling:
Abbott has responsibility for initial complaint registration.
Sentinel (the vendor) will investigate level II complaints.

4. MULTIGENT® Lithium Launch Strategy – Assay Disk
A disk will not be available at assay launch
Assay files must be entered manually using the package insert.
The MULTIGENT c Systems disk is targeted for 2nd quarter of 2008.
The disk will be available in conventional and SI units versions, however because of the prevalent use WW, Lithium assay units will be in SI units (mmol/L) on both disks.
No AEROSET disk will be produced.

5. Clinical Utility of Lithium
Used in the treatment of manic depressive psychosis. Elevated levels can cause intoxication. Early symptoms include apathy, sluggishness, drowsiness, lethargy, speech difficulties, tremor, muscle weakness and ataxia.
Long-term lithium therapy has been reported to cause hyperparathyroidism in some individuals with resulting hypercalcemia.

6. MULTIGENT® Lithium Methodology
Methodology: Colorimetric
CAP Code: 1428
Chemical Reaction:
Note: Schematic diagrams are for representative use only and do not reflect the actual chemical structure.

7. MULTIGENT® Lithium Methodology
Description of Chemical Reaction:
A substituted-porphyrin chromogen reacts with lithium at alkaline pH to form a Li+/chromogen complex. The resulting absorbance change is directly proportional to the concentration of lithium in the sample.
The formation of a complex between the chromogen and lithium is quite specific due to accommodation of the small ionic radius lithium ion (73 pm) within the chromogen core.
Formation of complexes between the chromogen and the larger sodium(113pm radius) and potassium (151 pm radius) ions have not been observed.
Li+/chromogen complex exhibits an absorbance maximum peak between 475nm and 495nm and an absorbance minimum between 505nm and 525nm.

8. MULTIGENT® Lithium Sample Handling
Specimen Requirements:
Serum
Plastic tubes
With or without gel barrier
Plasma
Sodium Heparin
K2-EDTA
Do NOT use Lithium Heparin
Note: Glass tubes and K3-EDTA were not tested.

9. MULTIGENT® Lithium Sample Handling
Sample Tubes tested and found acceptable:

10. MULTIGENT® Lithium 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 = 6 uL
System performs a Standard sample dilution of 1:20.17 using 6 uL of sample and 115 uL of saline diluent
An Automated Dilution protocol is available. The standard sample dilution is diluted 1:2 by using 3 uL of sample with 115 uL of saline diluent

11. MULTIGENT® Lithium Reagent Handling
Reagent is liquid, ready-to-use, single reagent kit.
Storage: 2 to 8°C.
Protect from light. 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 18 days.

12. MULTIGENT® Lithium Calibration
Calibration stability is 5 days
Uses MULTIGENT® Clin Chem Cal
Calibrator kits contain lithium and enzymatic creatinine values with the 2nd lot
Lithium reagent kit stuffer will contain lithium Clin Chem Cal values for previous lots of calibrator.
Information on calibration standardization is in the Calibration section of the Lithium package insert.

13. MULTIGENT ® Lithium Calibration Curve - ARCHITECT®
Note: Cal 1 concentration is automatically determined by the instrument dilution factor (1:20.17). The Calibrator concentration entered in the Configure assay parameters screen for this example is 1.52.

14. MULTIGENT ® Lithium Calibration Curve -AEROSET®

15. Quality Control
The following controls have been tested and initial control ranges have been assigned for AEROSET and ARCHITECT
Bio-Rad Liquid Assayed Multiqual Controls, Lot # 45540, Exp Date: 01/31/2009 and Lot # 45550, Exp. Date: 09/30/2009
Bio-Rad Lyphochek Assayed Chemistry Control, Lot # 14140, Exp Date: 12/31/2008 and Lot # 14150, Exp Date: 09/30/2009
Bio-Rad Lyphochek TDM Controls, Lot # 57160, Exp. Date: 6/30/2009 and Lot # 57170, Exp. Date: 10/31/2010

16. Quality Control
The Sentinel manufactured ClinChem Controls are available for use outside of the US.
These controls will not be launched for use with the U.S. Lithium kit.

17. MULTIGENT ® Lithium Reportable Range (Serum and Plasma)
Reportable Range = 0.10 to 3.51 mmol/L*
Linear within +/-5% or mmol/L whichever is greater
Limit of Quantitation = 0.10 mmol/L*
*Note: Alternate (conventional) units are mEQ/L which are
numerically equivalent to mmol/L.

18. MULTIGENT Lithium Interference
Note: A single spiked sample was used to test for Interference from Calcium, Copper, Iron, Magnesium, Potassium, Sodium and Zinc. That is why the Lithium concentration, bias and % Recovery are identical.

19. MULTIGENT® Lithium Precision
The precision of the Lithium assay is < 5% Total CV or < 0.075mmol/L SD, whichever is greater.
Representative data from a 20 day study based on Clinical and Laboratory Standards Institute (CLSI) protocol NCCLS EP5-A9 are summarized below.

20. MULTIGENT® Lithium Method Comparison
Results from the MULTIGENT Lithium assay on the ARCHITECT c 8000 System and the AEROSET System were compared with the results from the Trace Infinity Lithium Assay on a Hitachi 911. Results from the MULTIGENT Lithium assay on the ARCHITECT c 8000 System were compared with the results from atomic absorption spectrophotometry (AAS).

21. MULTIGENT® Lithium Method Comparison
Results from the MULTIGENT Lithium assay on the ARCHITECT c 8000 System and the ARCHITECT c System were compared with the results on the AEROSET System.

22. Lithium CAP Performance
CAP Proficiency Testing (PT) for MULTIGENT Lithium 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.
All samples were within 10% or 0.15 mmol/L as per our approved Product Requirements Document.
2007 CZ-B CAP Survey Results (Information Only) MULTIGENT All Method Mean
CHM
CHM CHM
CHM
CHM

23. ARCHITECT® Reaction graph (Low Range):

24. ARCHITECT® Reaction graph (High Range):

25. AEROSET® Reaction graph (Low Range):

26. AEROSET® Reaction graph (High Range):

27. Troubleshooting Tips Observed Problem: Elevated patient results
Probable Cause(s) CorrectiveAction(s)____
Use of Lithium heparin Use serum, Sodium heparin or EDTA
anticoagulant tubes blood collection tubes as described in the
reagent package insert.

28. Troubleshooting Tips
Observed Problem: Calibration stability does not meet labeling
Probable Cause(s) Corrective Action(s)____
Exposure of reagents to light Store unopened reagents in the original carton
until use. Keep reagent carousel covers closed
when not accessing reagents.

29. Questions and Answers
Why are there two separate list number products used for the US and ex-US?
Answer: More effective Distribution Strategy

30. Questions and Answers
Why won’t the Clin Chem Controls be approved for US use?
Answer: The number of users of Clin. Chem. Controls in the US would be very small, if at all.

31. Why do the reaction graphs for the low and high range samples progress in opposite directions?
Answer: Recall the reaction equation.
Note: Schematic diagrams are for representative use only and do not reflect the actual chemical structure.
Reaction of Free porphyrin with Lithium leads to complex formation and a shift of the spectrum characteristics of the free porphyrin.
The spectrum shift is characterized by a decrease of the Absorbance at 524 nm and by an increase at 476 nm.
At low lithium concentration the difference between Prim-Sec wavelengths produces positive values
At high lithium concentration the difference between the 2 wavelengths produces negative values

32. Why do the reaction graphs for the low and high range samples progress in opposite directions? (cont’d)
476 nm
524 nm
Note: Schematic reaction graphs are for representative use only, may not reflect the reaction in entirety.
The bold line depicts the absorbance response over time of lithium and porphryin in solution with no reaction taking place.
The dashed line depicts changes that take place over time during the reaction:
A positive absorbance change at 476 nm, the secondary wavelength with li-porphyrin complex formation
A negative absorbance change at 524 nm, the primary wavelength
A shift in the reaction graph towards the longer wavelength

33. Why do the reaction graphs for the low and high range samples progress in opposite directions? (cont’d)
The lithium reaction graphs plot the absorbance difference (Primary – Secondary) as a function of time.
Given the complexity of the Primary and Secondary wavelengths progressing in opposite directions, this difference has a downward trend at lower concentrations and an upward trend at higher concentrations.
Note: The reaction graphs show insignificant change in absorbance from reads 2-8, demonstrating > 90% reaction completion by the 2nd read.