AEROSET & ARCHITECT cSystems MULTIGENT DOA Assay Training for Phencyclidine (PCP) and Propoxyphene (Prpx) Urine Assays AEROSET PN No. 3.6.58 Version 2 Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Presentation Contents 9- Accuracy by Recovery 10- Cross Reactivity with other Drugs & Compounds 11- Method Comparison 12- Print Options 13- Assay Specific Information 14- Q & A 1- Basic Information 2- Ordering Information 3- Reaction Methodology 4- Sample Handling 5- Reagent Handling 6- Calibration 7- Quality Control 8- Precision Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 1 Basic Information Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

MULTIGENT PCP and Prpx DOA Assays Launch & Customer Support The MULTIGENT DOA assays are available for use on the AEROSET and cSystems (i.e. c4000, c8000, and c16000) Abbott will provide primary customer support, similar to all other Abbott Clinical Chemistry products. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Assay Disk / CD ARCHITECT cSystems assay disks containing the MULTIGENT DOA Assays are available for easy loading of the PCP and Prpx assays. There will be no AEROSET assay disk available. Assays can be manually configured. Perform assay configuration per the parameters provided in the assay specific application sheets. Assay Disks List Number ARCHITECT cSystems MULTIGENT Assay Disk Conventional / Alternate Units 8K50 ARCHITECT cSystems MULTIGENT Assay Disk SI / Alternate Units 8K51 ARCHITECT cSystems Special Chemistry Assay Disk Conventional / Alternate Units 2P10 ARCHITECT cSystems Special Chemistry Assay Disk 2P11 Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58

Reminder! Qualitative DOA Assays Qualitative DOA assays provide only a preliminary screening analytical test result. Qualitative results may be reported as the numeric value and/or as Positive / Negative. An alternate more specific analytical method must be used to obtain a confirmed result. Gas chromatography/mass spectrometry (GC/MS) is one well established confirmatory method. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Semiquantitative DOA Assays Semiquantitative (SQ) DOA assays still provide only a preliminary analytical test result. However, the result is reported as an approximate concentration, usually in ng/mL. Results are “semi”-quantitative because metabolites and drugs related to the drug used for calibration may not give responses identical to the calibrator compound. The semiquantitative result should still be confirmed since the identity of the detected drug or metabolite is often unknown. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 2 Ordering Information Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Ordering Information - Reagents *Package insert value reflects rounding “down” to 31 mL volume. Tests per kit is based on 31.7 mL fill volume. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Ordering Information Multiconstituent Calibrators I (DOA MC I) These Calibrators are used for PCP and Prpx. Each cal has a unique L/N and must be ordered separately. They do not come as a “set”. Note: SQ applications require use of all five levels. Qualitative applications require only: DOA MC Neg Cal and DOA MC I Cal 2. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Ordering Information - Controls PCP and Prpx use the DOA MC I Control set. Control Name List Number Config. Used for DOA MC I Control Set 3L43-10 2 x 5 mL Low 2 x 5 mL High Amphetamine/Methamphetamine, Barbiturates, Benzodiazepines, Cocaine, Opiates (when using the DOA MC I Cals), Methadone, Phencyclidine, Propoxyphene Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 3 Reaction Methodology Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reactions: Methodology is Enzyme Immunoassay (EIA) R1 reagent is added to specimen. R1 reagent contains Antibody to the drug of interest, NAD, and Glucose-6-Phosphate. Antibody binds to drug in the specimen. R2 reagent is added to the reaction mixture. R2 reagent contains Enzyme (G6PD)-drug conjugate. The enzyme-labeled drug competes for antibody with drug present in the sample. Antibody not bound to drug analyte in the specimen will bind to the conjugate. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reactions - DOAs (continued) When antibody binds to the conjugate, the conjugate enzyme’s (G6PD) active site is blocked and substrate (NAD) is not converted to product (NADH) and no absorbance is produced. (G6PD = Glucose-6-phosphate dehydrogenase) When there is more analyte (drug or metabolite) present in the specimen, there are more active conjugate enzymes (G6PD) available to convert substrate (NAD) to product (NADH). This results in a greater absorbance and therefore a higher result (analyte concentration). The methodology is exactly the same for the qualitative and semiquantitative applications. The difference between the qualitative and SQ applications is in the number of calibrators used. The qualitative applications use only two calibrators; the SQ applications use five calibrators. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reaction with R1 Antibody to the drug, NAD and Glucose-6-Phosphate in the R1 reagent is added to specimen and binds with the drug analyte contained in it. Antibody to drug + Drug Drug: Antibody complex (Sample) Also present in R1: NAD and Glucose-6-Phosphate Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reaction with R2 Enzyme-antigen conjugate (R2) is added to reaction mixture. Antibody not bound to drug analyte in the specimen will bind to the conjugate. This inactivates the G-6-PD. G-6-PD-Drug Conjugate + Drug:Antibody complex (R2) G-6-PD-Drug:Antibody complex (Enzyme becomes inactive) Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reaction: Detection of Drug Remaining ACTIVE Enzyme-antigen conjugate catalyzes the reduction of NAD to NADH. The resultant absorbance change is measured spectrophotometrically at 340 / 412 nm. G-6-PD-Drug Conjugate + NAD+Glucose-6-Phosphate (Active) NADH + 6-phosphogluconate Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reaction with little or no drug In the absence of drug in the sample, R1 antibody binds to R2 Enzyme-antigen conjugate. This forms an INACTIVE complex of antibody and enzyme labeled drug. NAD is not converted to NADH. Antibody to drug + G-6-PD-Drug (Active) (R1) (R2) G-6-PD-Drug:Antibody complex (Inactive) (G-6-PD = Glucose-6-phosphate dehydrogenase) Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reaction – Summary When analyte is present in the sample, enzyme activity is higher, resulting in a higher absorbance value. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 4 Sample Handling Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Sample Handling – Urine DOA assays Urine is the only acceptable specimen for PCP and Prpx The following materials are acceptable for urine collection containers: glass, plastic. Frozen specimens - thaw and mix well Turbid specimens – centrifuge as described in the package insert. Specimen storage 5 days stored frozen or refrigerated After 5 days, store frozen at -5° C or colder Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 5 Reagent Handling Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Reagent Handling MULTIGENT PCP and Prpx assays Supplied as liquid, ready-to-use, two reagent kit Storage must be at 2 to 8ºC Unopened Kit is stable to expiration date On board stability is 56 days Do not freeze or expose to temperatures above 32ºC Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 6 Calibration Not for customer distribution Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Calibration – Qualitative DOA Assays Calibration of the MULTIGENT DOA PCP and Prpx assays is performed in an identical manner to the other MULTIGENT DOA urine qualitative assays. The DOA MC Neg Cal is used as the BLANK. The DOA MC I Cal 2 is used as the CUTOFF calibrator. The package insert for the 3L43 Calibrators and Controls will be updated to include their use with the PCP and Prpx assays. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Calibrator Concentrations Calibrator L/N Designation Concentration PCP ng/mL Concentration Prpx ng/mL 3L43-05 Neg Cal 3L43-01 Cal 1 12.5 150 3L43-02 Cal 2 25 300 3L43-03 Cal 3 50 500 3L43-04 Cal 4 100 1000 Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

PCP & Prpx Qualitative Cutoff Summary Cutoff calibrator concentrations are listed below: Phencyclidine: 25 ng/mL (SAMHSA cutoff) Propoxyphene: 300 ng/mL Note that for both assays, Cal 2 contains the qualitative cutoff concentration Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Qualitative PCP: Two cal points AEROSET Calibration Qualitative PCP: Two cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Qualitative PCP: Two cal points c8000 Calibration Qualitative PCP: Two cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Qualitative Prpx: Two cal points AEROSET Calibration Qualitative Prpx: Two cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Qualitative Prpx: Two cal points c8000 Calibration Qualitative Prpx: Two cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Calibration – Semiquantitative Assays Semiquantitative applications are calibrated using the 5 calibrators specified in the assay specific package inserts. See also Section 13 of this presentation. "Assay Specific Information" Semiquantitative applications are calibrated in a manner identical to the other MULTIGENT DOA assays, or to other multi-calibrator assays such as the serum proteins. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Semiquantitative PCP: Five cal points AEROSET Calibration Semiquantitative PCP: Five cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Semiquantitative PCP: Five cal points c8000 Calibration Semiquantitative PCP: Five cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Semiquantitative Prpx: Five cal points AEROSET Calibration Semiquantitative Prpx: Five cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Semiquantitative Prpx: Five cal points c8000 Calibration Semiquantitative Prpx: Five cal points Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 7 Quality Control Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

DOA Quality Control: Qualitative Applications Positive and negative controls should be run at least once each day of assay use. Customers may wish to monitor the difference between control results and the cutoff concentration. Refer to page 11 of this presentation for information on MULTIGENT DOA MC I Control Products. If controls do not show the appropriate POS or NEG response, patient test results may be suspect and corrective action should be taken. Assay recalibration may be necessary. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Control Concentrations Control L/N Designation Target Values PCP ng/mL Target Values Prpx ng/mL 3L43-10 Ctrl 1 19 225 Ctrl 2 31 375 Controls are ordered as a set; both levels have the same L/N. Control target values are at +25% and -25% of the assay's cutoff. For PCP the values are rounded. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

DOA Quality Control: Semiquantitative Applications For SQ applications, customers should define ranges which include the nominal control concentrations provided in the package insert for the DOA MC I Calibrators and Controls. Controls should be run at least once each day of assay use. If controls values fall outside the established ranges, patient test results may be suspect and corrective action should be taken. Assay recalibration may be necessary. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 8 Precision Not for customer distribution Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Precision Claims for AEROSET and cSystems Qualitative Applications: Precision is < 7.5% Total CV for the qualitative PCP and Prpx assays. SQ Applications: Precision is < 10% Total CV for the SQ PCP and Prpx assays. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 9 Accuracy by Recovery Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Accuracy by Recovery Semiquantitative Applications Each SQ assay has been evaluated for accuracy by determining recovery of known analyte concentrations. In general, recovery was determined at calibrator concentrations and at concentrations at a mid-point between calibrators. Recovery values were between 90% and 110% with two values falling slightly outside this range, as shown below. Propoxyphene; 375 ng/mL Recovery = 113.8% Propoxyphene; 750 ng/mL Recovery = 111.1% The acceptance criterion for this protocol was recovery between 85% and 115%, so these results were acceptable. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 10 Cross Reactivity with other Drugs and Compounds Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Cross-Reacting Compounds Both the PCP and Prpx MULTIGENT DOA assays show some cross-reactivity with other drugs and compounds. Cross-reactivity is good when it permits detection of similar drugs, but it is bad when unrelated drugs may give false positive results. Both package inserts contain tables of drugs and other compounds that have been tested. Tables may give the concentration that reacts equivalently to the cutoff calibrator or a concentration that still shows a negative response. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 11 Method Comparison Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Method Comparison – Urine DOA Assays Package inserts contain method comparison data. The table below summarizes the %-agreement between AEROSET and the comparative device (A), and AEROSET vs. c 8000 (B) for the qualitative and semiquantitative assays. Qualitative Assay A B Phencyclidine 100 Propoxyphene Semiquantitative A B 100 99.1 Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 12 Print Options Not for customer distribution Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

MULTIGENT DOA Print Options Print options for the MULTIGENT PCP and Prpx DOA qualitative assays use two calibrators. Print options are not included in the package insert. They are provided on the application sheets with the assay specific parameters. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Print Options for AEROSET As shown in the DOA application sheets: Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Print Options for cSystems As shown in the DOA application sheets: Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Print Options: continued All qualitative values should be positive numbers, and are interpreted as a negative result only if the number is below the concentration of the cutoff calibrator. A negative result will NOT give a negative value. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 13 Assay Specific Information Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

MULTIGENT Phencyclidine Assay Method: enzyme immunoassay Reagent List#: 6L96-20 Calibrator List #: DOA MC Neg Cal: 3L43-05* DOA MC I Cal 1: 3L43-01 DOA MC I Cal 2: 3L43-02* DOA MC I Cal 3: 3L43-03 DOA MC I Cal 4: 3L43-04 Control List # DOA MC I Control Set: 3L43-10 Assay Stability On Board Stability: 56 Days Calibration Stability: 14 Days Assay Volumes Specimen: 12 uL R1 Reagent: 100 uL R2 Reagent: 100 uL Smartwashes: Required on AEROSET and cSystems for Amikacin LN 6L35, Digoxin LN 1E06, Digitoxin LN 6K40, Tobramycin LN 7F93, and Vancomycin LN 6E44. Required on cSystems for Gentamicin LN 1E11. Required on AEROSET for Direct Bilirubin LN 7D59. Microalbumin LN 2K98 requires smartwash for PCP on AEROSET. Qualitative Cutoff Conc: 25 ng/mL Semiquantitative Limit: 100 ng/mL Limit of Detection: 5 ng/mL Qualitative Precision < 7.5% Total CV Semiquantitative Precision < 10% Total CV * Calibrators used for the qualitative application Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

MULTIGENT Propoxyphene Assay Method: enzyme immunoassay Reagent List#: 6L97-20 Calibrator List #: DOA MC Neg Cal: 3L43-05* DOA MC I Cal 1: 3L43-01 DOA MC I Cal 2: 3L43-02* DOA MC I Cal 3: 3L43-03 DOA MC I Cal 4: 3L43-04 Control List # DOA MC I Control Set: 3L43-10 Assay Stability On Board Stability: 56 Days Calibration Stability: 14 Days Assay Volumes Specimen: 8 uL R1 Reagent: 100 uL R2 Reagent: 100 uL Smartwashes: Microalbumin LN 2K98 requires smartwash for Prpx on AEROSET. Qualitative Cutoff Conc: 300 ng/mL Semiquantitative Limit: 1,000 ng/mL Limit of Detection: 15 ng/mL Qualitative Precision < 7.5% Total CV Semiquantitative Precision < 10% Total CV * Calibrators used for the qualitative application Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Section 14 Q & A Not for customer distribution Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Questions and Answers How should qualitative controls be monitored? Some users may wish to monitor the difference between the control result and the cutoff calibration concentration. If the cutoff concentration is 300 ng/mL, and the negative control consistently gives values around 275, then this difference of 25 can provide a check of assay consistency. However, this difference will be affected by reagent lot, calibrator lot and control lot. There is no defined specification for this difference. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution

Not for customer distribution ** End of Document ** Questions and Answers Propoxyphene has calibrator levels that exceed 110% in the accuracy by recovery study? Are these levels considered out of specification? How can the data be explained? This is because these DOA assays are SEMI-quantitative. The calibration curve-fit is not as optimized as for a fully quantitative assay. Therefore, recovery values may vary in different regions of the cal curve. The assays still provide an approximate indication of drug concentration, which is what they are designed to do. Formal approval and revision history of this document can be found in the AEROSET Project Notebook, 3.6.58 Version 2 For Information Use Only, unless accompanied by associated electronic approval sheet from AEROSET Project Notebook, 3.6.58 Not for customer distribution ** End of Document **