Instructions for Use

2008-06

Microalbumin_2 (µALB_2)
System New Information
ADVIA® 1200 Updates to Reagents, Reagent Preparation and Use, Calibration
Frequency, Quality Control, Analytical Range, System Correlation
ADVIA 1650/1800 Updates to Reagents, Reagent Preparation and Use, Calibration
Frequency, Quality Control, Analytical Range, System Correlation
ADVIA 2400 Updates to Reagents, Reagent Preparation and Use, Calibration
Frequency, Quality Control, Analytical Range, System Correlation

NOTE: For an explanation of the symbols in this document and associated products,
refer to Understanding the Symbols located on the introductory page of the Methods
Directory.

Method Summary
Item Description
Method Principle PEG-enhanced immunoturbidimetric
Specimen Type Human urine
On-board Stability Without
Reagent With Reagent
ADVIA Chemistry Container Container
Systems Insert Insert
ADVIA 1200 60 days 60 days
ADVIA 1650/1800 60 days 60 days
ADVIA 2400 60 days 60 days
Reagent Storage Temperature 2–8°C
Calibration Frequency Without
Reagent With Reagent
ADVIA Chemistry Container Container
Systems Insert Insert
ADVIA 1200 20 days 30 days
ADVIA 1650/1800 60 days 60 days
ADVIA 2400 20 days 60 days
Reagent Blank (RBL) Frequency At time of method calibration
Reaction Type Endpoint (EPA)
Measurement Wavelength 340 nm
Standardization Internal standard

© 2007 Siemens Medical Solutions Diagnostics. All rights reserved.

Siemens Medical Solutions Diagnostics, Tarrytown, NY 10591-5097 USA
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Item Description
Analytical Range Urine Specimen*
0.3–(38.0–42.0) mg/dL
(3–[380–420]) mg/L
*The albumin (ALB) concentration in the
Microalbumin_2 Calibrators, Level 5, varies from
38.0–42.0 mg/dL
(380–420 mg/L).
Expected Values < 30 mg/day

Reagent Code 74090

Calibrator ADVIA Chemistry Microalbumin_2 Calibrators:
REF 06487733

Intended Use
For in vitro diagnostic use in the quantitative determination of albumin in human urine
on the ADVIA Chemistry systems. Such measurements are used in the diagnosis and
treatment of microalbuminuria and are helpful for the detection and treatment of
patients at risk from renal disease.

Summary and Explanation 1,2,3,6

The Microalbumin_2 (µALB_2) method is based on the work of Fielding and Hellsing,
and it measures very small levels of albumin in urine samples. Albumin is a plasma
protein that is responsible for much of the osmotic force of the blood. In a healthy
population, only a small amount of albumin (up to 30 mg/day) is excreted in the urine.
Elevated levels of urinary albumin indicate a high probability of damage of the
glomerular filtration capacity of the kidney. During the progression of renal disease in
type I diabetes mellitus, stage III or incipient nephropathy is characterized by the
elevation in urinary albumin. Elevated results in urinary albumin may also be
associated with hypertension, some lipid abnormalities, and several immune disorders
as well as other conditions such as vigorous exercise, blood in the urine, urinary tract
infection, dehydration, and some drugs.

Principles of the Procedure

The microalbumin method is a PEG enhanced immunoturbidimetric assay. A sample
containing human albumin is suitably diluted and then reacted with specific antiserum
to form a precipitate that can be measured turbidimetrically at 340 nm. By constructing
a standard curve from the absorbencies of standards, the albumin concentration of the
sample is determined.

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Reagents
The reagents are packaged as listed below. Components of the package are available
as a kit only.

REF
Container Size Symbol Contents Amount No. of Tests
03051194 Microalbumin 4 x 105
Reagents (ADVIA
1650/1800)
20-mL Reagent 1 4 x 20 mL 4 x 130
(ADVIA
1200/2400)
20-mL Reagent 2 4 x 4.3 mL

Components and Concentrations

Reagent Component Concentration

Reagent 1 Polyethylene glycol 6.00%
Sodium azide 0.09%
Reagent 2 Antihuman albumin (goat) lot-specific
Sodium azide 0.09%

CAUTION! This device contains material of animal origin and should be handled as a
potential carrier and transmitter of disease.

NOTE: Sodium azide can react with copper and lead plumbing to form explosive metal
azides. If disposal into a drain is in compliance with federal, state, and local
requirements, flush reagents with a large amount of water to prevent the buildup of
azides.

For In Vitro Diagnostic Use.

Reagent Preparation and Use

Reagents are ready to use. Before use, gently swirl the reagent to disrupt bubbles and
assure homogeneity. If bubbles still exist or foam is present, using a clean transfer
pipette, aspirate them from the reagent container prior to use.

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On-board Reagent Stability (OBS)

Stability without Reagent Stability with Reagent
System Container Insert Container Insert
ADVIA 1200 60 days 60 days
ADVIA 1650/1800 60 days 60 days
ADVIA 2400 60 days 60 days

For all systems, unopened reagents are stable until the expiration date printed on the
product label when stored at 2–8°C. Do not freeze reagents.

For additional details, refer to the Methods Introduction section of the system-specific
Operator’s Guide.

Sample Handling
We recommend using urine for this method. Collect the urine as a 24-hour, timed, or
random midstream sample (spot collection) in a clean, unused glass or plastic
collection container. Preservatives are not recommended for this method.

Specimen Storage 3
For timed or 24-hour samples, if possible, use fresh specimens. You can also use
samples that have been stored up to 2 weeks at 4°C or up to 5 months at -70°C. Avoid
repeated freeze-thaw cycles.

For additional details, refer to Sample Collection and Preparation in the Methods
Introduction section of the system-specific Operator’s Guide.

For instructions on how to load reagents and run samples, refer to the Daily Operations
section of the system-specific Operator’s Guide.
NOTE: Mix stored specimens adequately and centrifuge prior to testing.

Materials Required but not Provided

The following list contains the materials required, but not provided, to perform this
method:
• sample containers
• system solutions
• calibrator and control materials (refer to the Method Summary section for the REFs)
• reagent container adapters:
• 20-mL adapter (REF 02404085; PN 094-0159-01) for 40-mL slot
(ADVIA 1200/1800)
• 20-mL adapter (REF 05249323; PN 073-0936-01) for 70-mL slot (ADVIA 1800)
• 20-mL adapter (REF 00771668; PN 073-0345-02) for 70-mL slot
(ADVIA 1650/2400)
• reagent container inserts (REF 02991886)

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Calibration
Calibrate this method using ADVIA Chemistry Microalbumin_2 Calibrators
(REF 06487733) as follows:
1. Refer to the package insert supplied with the calibrator for handling instructions
and values.
2. Place 1 sample container filled with distilled water as the blank sample onto the
sample tray (SST) cup position assigned to the blank for multipoint calibration.
3. At the Setup menu, select the Con/Cal Sample Setup window.
4. Enter the expected sample container type and assigned SST cup position.
5. If no sample type and cup position are defined, you must do so before running the
calibration.
6. Place each of the 5 levels of the Microalbumin Calibrator onto the SST cup position
assigned for each level.
For setup and use instructions, refer to the Calibration Overview section of the
system-specific Operator’s Guide.

Calibration Frequency
Perform a calibration when this method is implemented on the system. You must
recalibrate after the following events:
• when the reagent lot number changes
• after replacing critical optical or hydraulic components
• when indicated by quality control procedures

Siemens has validated the calibration stability for this method as shown in the following
table:

Minimum Calibration Minimum Calibration

Frequency without Reagent Frequency with Reagent
ADVIA Chemistry System Container Insert Container Insert
ADVIA 1200 20 days 30 days
ADVIA 1650/1800 60 days 60 days
ADVIA 2400 20 days 60 days

Siemens recommends calibrating new reagent packs if the previous reagent pack was
calibrated any time during its on-board stability, other than as a fresh pack.

Individual laboratory quality control programs and procedures may require more
frequent calibration.

Reagent Blank (RBL) Frequency

The RBL is measured at the time of method calibration.

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Quality Control
Siemens recommends the use of quality control material from Bio-Rad Laboratories
with at least 2 levels (low and high). A satisfactory level of performance is achieved
when the analyte values obtained are within the Acceptable Control Range for the
system or within your range, as determined by an appropriate internal laboratory
quality control scheme.

The actual frequency of control in a laboratory is based on many factors, such as

workflow, system experience, and government regulation. Each laboratory should
evaluate the controls based on the frequency established by their laboratory
guidelines. When the method is performed, analyze at least 2 levels of controls daily.

Also, assay controls under the following conditions:

• whenever you use a new reagent lot
• following the performance of any system maintenance, cleaning, or troubleshooting
procedure
• after performing a new calibration

For more information, refer to the Quality Control Overview section of the
system-specific Operator’s Guide.

Limitations of the Procedure 4

A number of substances cause physiological changes in urine analyte concentrations.
A comprehensive discussion of possible interfering substances, their urine
concentrations, and their possible physiological involvements is beyond the scope of
this document. Consult listed reference for specific details on known potential
interfering substances.4

As with any chemical reaction, you must be alert to the possible effect on results of
unknown interferences from medications or endogenous substances. The laboratory
and physician must evaluate all patient results in light of the total clinical status of the
patient.

Interference
Siemens tested the following potential interferents with a urine pool of approximately
3 mg/dL albumin concentration and found the results shown below:

ADVIA Chemistry Systems

µALB_2 Sample
Interferent Interferent Level Concentration Interference*

Acetaminophen 100 mg/dL 3.0 mg/dL NSI

(0.66 mmol/L) (30 mg/L)
Ascorbic Acid 500 mg/dL 3.0 mg/dL NSI
(28.4 mmol/L) (30 mg/L)
Calcium 400 mg/dL 3.0 mg/dL NSI
(100 mmol/L) (30 mg/L)
Citrate 500 mg/dL 3.0 mg/dL NSI
(26 mmol/L) (30 mg/L)

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Microalbumin_2 (µALB_2)

µALB_2 Sample
Interferent Interferent Level Concentration Interference*

Creatinine 500 mg/dL 3.0 mg/dL NSI

(44.2 mmol/L) (30 mg/L)
Glucose 5000 mg/dL 3.0 mg/dL NSI
(277.5 mmol/L) (30 mg/L)
Hemoglobin 500 mg/dL 3.0 mg/dL NSI
(5.0 g/L) (30 mg/L)
Hippuric Acid 400 mg/dL 3.0 mg/dL NSI
(22.3 mmol/L) (30 mg/L)
Inorganic Phosphorous 400 mg/dL 3.0 mg/dL NSI
(129.2 mmol/L) (30 mg/L)

Magnesium 400 mg/dL 3.0 mg/dL NSI

(42.0 mmol/L) (30 mg/L)

Oxalate 30 mg/dL 3.0 mg/dL NSI

(3.42 mmol/L) (30 mg/L)
Potassium Chloride 1000 mg/dL 3.0 mg/dL NSI
(134.1 mmol/L) (30 mg/L)
Salicylate 250 mg/dL 3.0 mg/dL NSI
(18.1 mmol/L) (30 mg/L)
Sodium Chloride 2000 mg/dL 3.0 mg/dL NSI
(342.2 mmol/L) (30 mg/L)
Urea Nitrogen 400 mg/dL 3.0 mg/dL NSI
(142.8 mmol/L) (30 mg/L)
Uric Acid 100 mg/dL 3.0 mg/dL NSI
(5.9 mmol/L) (30 mg/L)
*NSI = No Significant Interference. A percentage effect of 10% is considered a significant
interference.

Performance Characteristics
Precision 5
Each sample was assayed 2 times per run, 2 runs per day, for at least 10 days.
Precision estimates were computed according to CLSI document EP05-A2, Evaluation
of Precision Performance of Quantitative Measurement Methods; Approved Guideline.5

Data contained in this section represents typical performance for

ADVIA Chemistry systems. Your laboratory data may differ from these values.

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Microalbumin_2 (µALB_2)

Conversion factor: mg/dL x 10 = mg/L

ADVIA 1200
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)
Common Units (mg/dL)
Pool 1 2.9 0.06 2.2 0.11 3.7
Pool 2 29.8 0.41 1.4 0.53 1.8
Control 1 2.4 0.09 3.8 0.11 4.7
Control 2 10.4 0.14 1.3 0.19 1.8
SI Units (mg/L)
Pool 1 29 0.6 2.2 1.1 3.7
Pool 2 298 4.1 1.4 5.3 1.8
Control 1 24 0.9 3.8 1.1 4.7
Control 2 104 1.4 1.3 1.9 1.8

ADVIA 1650/1800
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)
Common Units (mg/dL)
Pool 1 2.9 0.06 2.1 0.10 3.3
Pool 2 29.3 0.34 1.2 0.49 1.7
Control 1 1.9 0.08 4.1 0.12 6.1
Control 2 9.1 0.16 1.8 0.23 2.5
SI Units (mg/L)
Pool 1 29 0.6 2.1 1.0 3.3
Pool 2 293 3.4 1.2 4.9 1.7
Control 1 19 0.8 4.1 1.2 6.1
Control 2 91 1.6 1.8 2.3 2.5

ADVIA 2400
Within-Run Total
Specimen Type Level SD CV (%) SD CV (%)
Common Units (mg/dL)
Pool 1 2.8 0.05 1.6 0.08 2.9
Pool 2 28.8 0.24 0.8 0.28 1.0
Control 1 1.7 0.04 2.1 0.07 3.8
Control 2 8.9 0.08 0.9 0.10 1.1
SI Units (mg/L)
Pool 1 28 0.5 1.6 0.8 2.9
Pool 2 288 2.4 0.8 2.8 1.0
Control 1 17 0.4 2.1 0.7 3.8
Control 2 89 0.8 0.9 1.0 1.1

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Microalbumin_2 (µALB_2)

Analytical Range
This method measures the ALB concentration in urine ranging from the minimum
detectable concentration (MDC) to the ALB concentration in the highest level of the
calibrator according to the table shown below. MDC is an estimation based on two
times the within-run standard deviation of the zero calibrator.

System Urine Specimen*

ADVIA 1200 0.3– (38.0–42.0) mg/dL

(3– [380–420] mg/L)
ADVIA 1650/1800 0.3– (38.0–42.0) mg/dL
(3– [380–420] mg/L)
ADVIA 2400 0.3– (38.0–42.0) mg/dL
(3– [380–420] mg/L)
*the ALB concentration in the Microalbumin_2 Calibrator Level 5
varies from 38.0 to 42.0 mg/dL (380 to 420 mg/L).

Siemens has validated an automatic rerun condition for this method that extends the
reportable ranges for urine up to 380 mg/dL (3800 mg/L) for all ADVIA Chemistry
systems.

Prozone Effect
A prozone effect was not demonstrated for analyte concentrations up to at least
20,000 mg/dL (200,000 mg/L) for all ADVIA Chemistry systems.

Expected Values 6
The expected value for this method is < 30 mg/day for urine.

We provide this information for reference. Each laboratory should establish its own
normal range. You can enter normal range values and abnormal range values at the
Analytical Parameters (Chemistry) window.

System Correlation
The performance of the applicable method (y) was compared with the performance of
the same method on the comparison system (x).

ADVIA 1200
Specimen Type Comparison System (x) N Regression Equation Sy.x r Sample Range
Urine ADVIA 1650/1800 95 y = 0.97x - 0.1 0.4 0.996 0.6–16.7 mg/dL
µALB_2 y = 0.97x - 0.7 4.4 0.996 6–167 mg/L
Urine ADVIA 1200 µALB 96 y = 0.99x - 0.1 0.2 0.999 0.8–16.9 mg/dL
y = 0.99x - 1.0 2.2 0.999 8–169 mg/L

ADVIA 1650/1800
Specimen Type Comparison System (x) N Regression Equation Sy.x r Sample Range
Urine ADVIA 1650/1800 98 y = 0.99x - 0.1 0.1 0.999 0.6–16.8 mg/dL
µALB y = 0.99x - 0.7 1.2 0.999 6–168 mg/L

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ADVIA 2400
Specimen Type Comparison System (x) N Regression Equation Sy.x r Sample Range
Urine ADVIA 1650/1800 95 y = 0.97x - 0.0 0.4 0.997 0.6–16.7 mg/dL
µALB_2 y = 0.97x - 0.0 3.5 0.997 6–167 mg/L
Urine ADVIA 2400 µALB 97 y = 1.00x - 0.2 0.6 0.991 0.5–17.1 mg/dL
y = 1.00x - 2.4 6.4 0.991 5–171 mg/L

Standardization
The ADVIA µALB_2 method is traceable to an internal standard manufactured using
highly purified material. Currently, no reference standard for this method exists.
Assigned values of ADVIA Chemistry Microalbumin_2 Calibrators are traceable to this
standardization.

References
1. Fielding BA, Price DA, Houlton CA. Enzyme immunoassay for urinary albumin. Clin
Chem.1983;29:355-357.
2. Hellsing K. Influenced polymers on the antigen-antibody reaction in a continuous
flow system. In: Automated Immunoprecipitin Reactions. Colloquium on AIP.
Tarrytown, NY: Technicon Inst. Corp.; 1972:17, 798-9.
3. Burtis CA, Ashwood ER, Bruns DE. Tietz Textbook of Clinical Chemistry and
Molecular Diagnostics, 4th edition. St. Louis, MO.: Elsevier Saunders;
2006:886-888.
4. Young DS. Effects of Drugs on Clinical Laboratory Tests. 5th ed. Washington:
AACC Press (2000).
5. Clinical and Laboratory Standards Institute (formerly NCCLS). Evaluation of
Precision Performance of Quantitative Measurement Methods; Approved Guideline
- Second Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2004.
NCCLS Document EP05-A2.
6. Wu AHB. Tietz Clinical Guide to Laboratory Tests, 4th edition, Saunders Elsevier,
St. Louis, MO: 2006:316.

Technical Assistance
For customer support, please contact your local technical support provider or
distributor.

Trademarks
ADVIA is a trademark of Siemens Medical Solutions Diagnostics.

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Siemens Medical Solutions

Diagnostics Pty Ltd
ABN 65 007 436 651
885 Mountain Highway
Bayswater Victoria 3153
Australia

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