ADVIA Centaur ® CP

Immunoassay System

Prolactin (PRL)
Assay Summary
Sample Type Serum
Sample Volume 25 µL
Calibrator B
Sensitivity and Assay Range 0.3 – 200 ng/mL (6.4 – 4240 µIU/mL)

Contents
REF Contents Number of Tests
09505871 5 ReadyPack® primary reagent packs containing ADVIA Centaur® 250
(110758) PRL Lite Reagent and Solid Phase
ADVIA Centaur and ADVIA Centaur CP PRL Master Curve cards
or
03278474 1 ReadyPack primary reagent pack containing ADVIA Centaur 50
(110757) PRL Lite Reagent and Solid Phase
ADVIA Centaur and ADVIA Centaur CP PRL Master Curve cards

Intended Use
For in vitro diagnostic use in the quantitative determination of prolactin in serum using the
ADVIA Centaur CP System.

Materials Required But Not Provided

REF Description Contents
00652707 Calibrator B 6 vials of low calibrator
6 vials of high calibrator
or
00649625 Calibrator B 2 vials of low calibrator
2 vials of high calibrator

Optional Reagents
REF Description Contents
07907174 ADVIA Centaur Multi-Diluent 1 6 ReadyPack ancillary reagent packs containing
(110313) 25 mL/pack
or
07293184 ADVIA Centaur Multi-Diluent 1 2 ReadyPack ancillary reagent packs containing
(110312) 25 mL/pack
09348792 Multi-Diluent 1 50 mL/vial
(672177)
08192578 PRL Master Curve Material 10 x 1 mL
(672430)

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PRL ADVIA Centaur CP System

Summary and Explanation of the Test

Prolactin is a single-chain polypeptide hormone secreted by the anterior pituitary1 under the
control of prolactin-inhibiting factors and prolactin-releasing factors. These inhibiting and
releasing factors are secreted by the hypothalamus.2 Prolactin is also synthesized by the
placenta and is present in amniotic fluid.
Prolactin initiates and maintains lactation in females.3 It also plays a role in regulating gonadal
function in both males and females. In adults, basal circulating prolactin is present in
concentrations up to 30 ng/mL (636 μIU/mL). During pregnancy and postpartum lactation,
serum prolactin can increase 10- to 20-fold. Exercise, stress, and sleep also cause transient
increases in prolactin levels.
Consistently elevated serum prolactin levels greater than 30 ng/mL (636 μIU/mL) in the
absence of pregnancy and postpartum lactation are indicative of hyperprolactinemia, which is
the most common hypothalamic-pituitary dysfunction encountered in clinical endocrinology.
Hyperprolactinemia often results in galactorrhea, amenorrhea, and infertility in females, and in
impotence and hypogonadism in males.4 Renal failure, hypothyroidism, and prolactin-
secreting pituitary adenomas are also common causes of abnormally elevated prolactin
levels.1,5,6

Assay Principle
The ADVIA Centaur CP Prolactin assay is a two-site sandwich immunoassay using direct
chemiluminometric technology, which uses constant amounts of two antibodies. The first
antibody, in the Lite Reagent, is a polyclonal goat anti-prolactin antibody labeled with
acridinium ester. The second antibody, in the Solid Phase, is a monoclonal mouse anti-
prolactin antibody, which is covalently coupled to paramagnetic particles.
The system automatically performs the following steps:
• dispenses 25 μL of sample into a cuvette
• dispenses 100 μL of Lite Reagent and incubates for 6.3 minutes at 37°C
• dispenses 450 μL of Solid Phase and incubates for 3.0 minutes at 37°C
• separates, aspirates, and washes the cuvettes with Wash 1
• dispenses 300 μL each of Acid Reagent (R1) and Base Reagent (R2) to initiate the
chemiluminescent reaction
• reports results according to the selected option, as described in the system operating
instructions or in the online help system
A direct relationship exists between the amount of prolactin present in the patient sample and
the amount of relative light units (RLUs) detected by the system.

Specimen Collection and Handling

Serum is the recommended sample type for this assay.
The following recommendations for handling and storing blood samples are furnished by the
Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS)7
• Collect all blood samples observing universal precautions for venipuncture.
• Allow samples to clot adequately before centrifugation.
• Keep tubes stoppered and upright at all times.
• Do not use samples that have been stored at room temperature for longer than 8 hours.
• Tightly cap and refrigerate specimens at 2 to 8°C if the assay is not completed within
8 hours.

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• Freeze samples at or below -20°C if the sample is not assayed within 48 hours.
• Freeze samples only once and mix thoroughly after thawing.
Before placing samples on the system ensure that:
• Samples are free of fibrin or other particulate matter.
• Samples are free of bubbles.

Reagents
Store the reagents upright at 2–8°C.
Mix all primary reagent packs by hand before loading them onto the system. Visually inspect the bottom of the
reagent pack to ensure that all particles are dispersed and resuspended. For detailed information about
preparing the reagents for use, refer to the Operator’s Guide.

Reagent Pack Reagent Volume Ingredients Storage Stability

ADVIA Centaur Lite Reagent 5.0 mL/ polyclonal goat anti-prolactin 2–8°C until the expiration date
PRL ReadyPack reagent antibody (~0.16 μg/mL) on the pack label.
primary reagent pack labeled with acridinium ester For onboard stability,
pack in buffer with sodium azide refer to Onboard Stability
(0.11%) and preservatives and Calibration Interval.
Solid Phase 22.5 mL/ monoclonal mouse anti- 2–8°C until the expiration date
reagent prolactin antibody on the pack label.
pack (~3.67 μg/mL) covalently For onboard stability,
coupled to paramagnetic refer to Onboard Stability
particles in buffer with protein and Calibration Interval.
stabilizers, sodium azide
(0.11%), and preservatives
ADVIA Centaur Multi-Diluent 1 25.0 mL/ equine serum with sodium 2–8°C until the expiration date
reagent azide (0.1%) and preservatives on the pack label
ReadyPack pack or
ancillary reagent 28 consecutive days after
pack accessing the ancillary
reagent pack
Safety data sheets (MSDS/SDS) available on www.siemens.com/diagnostics.
R22 Harmful! Harmful if swallowed. After contact with skin, wash immediately with plenty of soap
S28 and water. Contains: sodium azide; Lite Reagent, Solid Phase

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.
On disposal, flush reagents with a large volume of water to prevent the buildup of azides, if
disposal into a drain is in compliance with federal, state, and local requirements.
For In Vitro Diagnostic Use.

Loading Reagents
Ensure that the system has sufficient primary and ancillary reagent packs. For detailed
information about preparing the system, refer to the system operating instructions or to the
online help system.
Mix all primary reagent packs by hand before loading them onto the system. Visually inspect
the bottom of the reagent pack to ensure that all particles are dispersed and resuspended. For
detailed information about preparing the reagents for use, refer to the Operator’s Guide.

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PRL ADVIA Centaur CP System

Load the primary reagent packs in the primary reagent area. The arrows on the end label can
be used as a placement guide. However left, center, and right placement of the primary reagent
packs is not required because there is only one reagent probe on the ADVIA Centaur CP
System. The system automatically mixes the primary reagent packs to maintain homogeneous
suspension of the reagents. For detailed information about loading reagents, refer to the
system operating instructions or to the online help system.
If automatic dilution of a sample is required, load ADVIA Centaur Multi-Diluent 1 in the
ancillary reagent area.

Onboard Stability and Calibration Interval

Onboard Stability Calibration Interval
28 days 28 days

Additionally, the ADVIA Centaur CP Prolactin assay requires a two-point calibration:

• when changing lot numbers of primary reagent packs
• when replacing system components
• when quality control results are repeatedly out of range
NOTE:
• Discard the primary reagent packs at the end of the onboard stability interval.
• Do not use reagents beyond the expiration date.

Master Curve Calibration

The ADVIA Centaur CP Prolactin assay requires a Master Curve calibration when using a
new lot number of Lite Reagent and Solid Phase. For each new lot number of Lite Reagent and
Solid Phase, use the barcode reader or keyboard to enter the Master Curve values on the
system. The Master Curve card contains the Master Curve values. For detailed information
about entering calibration values, refer to the system operating instructions or to the online
help system.

Quality Control
Follow government regulations or accreditation requirements for quality control frequency.
For detailed information about entering quality control values, refer to the system operating
instructions or to the online help system.
To monitor system performance and chart trends, as a minimum requirement, two levels of
quality control material should be assayed on each day that samples are analyzed. Quality
control samples should also be assayed when performing a two-point calibration. Treat all
quality control samples the same as patient samples.
Siemens Healthcare Diagnostics recommends the use of commercially available quality
control materials 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.
If the quality control results do not fall within the Expected Values or within the laboratory’s
established values, do not report results. Take the following actions:
• Verify that the materials are not expired.
• Verify that required maintenance was performed.
• Verify that the assay was performed according to the instructions for use.

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• Rerun the assay with fresh quality control samples.
• If necessary, contact your local technical support provider or distributor for assistance.

Sample Volume
This assay requires 25 μL of sample for a single determination. This volume does not include
the unusable volume in the sample container or the additional volume required when
performing duplicates or other tests on the same sample. For detailed information about
determining the minimum required volume, refer to the system operating instructions or to the
online help system.
NOTE: The sample volume required to perform onboard dilution differs from the sample
volume required to perform a single determination. Refer to the following information for the
sample volume required to perform onboard dilutions:
Dilution Sample Volume (µL)
1:2 100
1:5 40

Assay Procedure
For detailed procedural information, refer to the system operating instructions or to the online
help system.

Procedural Notes
Calculations
For detailed information about how the system calculates results, refer to the system operating
instructions or to the online help system.
The system reports serum prolactin results in ng/mL (common units) or μIU/mL (SI units),
depending on the units defined when setting up the assay. The conversion formula is
1 ng/mL = 21.2 μIU/mL.

Dilutions
• Serum samples with prolactin levels greater than 200 ng/mL (4240 μIU/mL) must be
diluted and retested to obtain accurate results.
• Patient samples can be automatically diluted by the system or prepared manually.
• For automatic dilutions, ensure that ADVIA Centaur Multi-Diluent 1 is loaded and set the
system parameters as follows:
Dilution point: ≤ 200 ng/mL (4240 μIU/mL)
Dilution factor: 2, 5
For detailed information about automatic dilutions, refer to the system operating
instructions or to the online help system.
• Manually dilute the patient samples when patient results exceed the linearity of the assay
using automatic dilution, or when laboratory protocol requires manual dilution.
• Use Multi-Diluent 1 to manually dilute patient samples, and then load the diluted sample
in the sample rack, replacing the undiluted sample.
• Ensure that results are mathematically corrected for dilution. If a dilution factor is entered
when scheduling the test, the system automatically calculates the result.

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PRL ADVIA Centaur CP System

High Dose Hook Effect

Patient samples with high prolactin levels can cause a paradoxical decrease in the RLUs (high
dose hook effect). In this assay, patient samples with prolactin levels as high as 30,000 ng/mL
(636,000 μIU/mL) will assay greater than 200 ng/mL (4240 μIU/mL).

Disposal
Dispose of hazardous or biologically contaminated materials according to the practices of your
institution. Discard all materials in a safe and acceptable manner, and in compliance with all
federal, state, and local requirements.

Limitations
Pregnancy, lactation, and the administration of oral contraceptives can increase prolactin
concentrations.1
Heterophilic antibodies in human serum can react with reagent immunoglobulins, interfering
with in vitro immunoassays.8 Patients routinely exposed to animals or to animal serum
products can be prone to this interference and anomalous values may be observed. Additional
information may be required for diagnosis.
Serum specimens that are . . . Demonstrate ≤ 5% change in results up to . . .
hemolyzed 500 mg/dL of hemoglobin
lipemic 1000 mg/dL of triglycerides
icteric 20 mg/dL of bilirubin

Interference testing was determined according to CLSI Document EP7-A2.9

Expected Results
The expected results for the ACS:180® Prolactin assay were previously established. Data was
obtained on serum samples from 661 apparently healthy individuals. Based on a central 95%
interval, the following reference ranges were established:
Mean Range Mean Range
Category N (ng/mL) (ng/mL) (µIU/mL) (µIU/mL)
Females
Nonpregnant 202 9.6 2.8–29.2 204 59–619
Pregnant 216 61.7 9.7–208.5 1308 206–4420
Postmenopausal 104 6.9 1.8–20.3 146 38–430
Males 139 7.0 2.1–17.7 148 45–375

These results were confirmed for the ADVIA Centaur CP Prolactin assay by analyzing 119
samples in the range of 2.2 to 147.8 ng/mL (46.6 to 3133.4 μIU/mL). Refer to Method
Comparison.
As with all in vitro diagnostic assays, each laboratory should determine its own reference
range(s) for the diagnostic evaluation of patient results.10

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Performance Characteristics
Specificity
The cross-reactivity of the assay with TSH, LH, hCG, FSH, hGH, and hPL was determined by
adding these hormones to serum samples containing prolactin. The level of prolactin in the
samples was then determined.
Prolactin value without cross-reactant Prolactin value with cross-reactant
Cross-reactant (ng/mL) (µIU/mL) (ng/mL) (µIU/mL)
TSH; 1000 μIU/mL 8.16 173 8.55 181
42.82 908 43.40 920
109.68 2325 107.30 2275
LH; 250 mIU/mL 8.16 173 8.84 187
42.82 908 43.13 914
109.68 2325 104.88 2223
hCG; 200,000 mIU/mL 8.16 173 8.30 176
42.82 908 40.28 854
109.68 2325 109.60 2324
FSH; 250 mIU/mL 8.16 173 8.40 178
42.82 908 42.66 904
110.08 2325 105.05 2227
hGH; 500 ng/mL 4.51 96 4.56 97
42.82 908 42.22 895
109.68 2325 108.22 2294
hPL; 12.5 μg/mL 2.50 53 2.51 53
35.13 745 36.40 772
88.67 1880 88.18 1869
Interference testing was previously determined for the ACS:180 Prolactin assay.
Sensitivity and Assay Range
The ADVIA Centaur CP Prolactin assay measures prolactin concentrations up to 200 ng/mL
(4240 μIU/mL) with a minimum detectable concentration (analytical sensitivity) of 0.3 ng/mL
(6.4 μIU/mL). Analytical sensitivity is defined as the concentration of prolactin that
corresponds to the RLUs that are two standard deviations greater than the mean RLUs of
20 replicate determinations of the prolactin zero standard.
Method Comparison
For 119 samples in the range of 2.2 to 147.8 ng/mL (46.6 to 3133.4 μIU/mL), the relationship
between the ADVIA Centaur CP Prolactin assay and the ACS:180 Prolactin assay is described
by the equation:
ADVIA Centaur CP Prolactin = 0.98 (ACS:180 Prolactin) – 0.17 ng/mL
Correlation coefficient (r) = 0.993
For 119 samples in the range of 3.3 to 142.9 ng/mL (70.0 to 3029.5 μIU/mL), the relationship
between the ADVIA Centaur CP Prolactin assay and the ADVIA Centaur Prolactin assay is
described by the equation:
ADVIA Centaur CP Prolactin = 0.98 (ADVIA Centaur Prolactin) - 0.27 ng/mL
Correlation coefficient (r) = 0.995

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Dilution Recovery
Six human serum samples in the range of 104.2 to 166.0 ng/mL (2208.8 to 3518.8 μIU/mL) of
prolactin were diluted 1:2, 1:4, 1:8, and 1:16 with Multi-Diluent 1 and assayed for recovery
and parallelism. The recoveries ranged from 95.5% to 111.1% with a mean of 104.6%.
Observed Expected Observed Expected
Sample Dilution (ng/mL) (ng/mL) (µIU/mL) (µIU/mL) Recovery %
1 — 104.2 104.2 2208.8 2208.8
1:2 53.9 52.1 1142.0 1104.4 103.4
1:4 27.2 26.0 576.5 552.2 104.4
1:8 13.9 13.0 294.2 276.1 106.5
1:16 7.2 6.5 153.4 138.1 111.1
Mean 106.4
2 — 123.7 123.7 2622.5 2622.4
1:2 65.1 61.9 1380.2 1311.2 105.3
1:4 32.0 30.9 679.3 655.6 103.6
1:8 14.8 15.5 313.1 327.8 95.5
1:16 7.8 7.7 165.2 163.9 100.8
Mean 101.3
3 — 121.5 121.6 2576.8 2576.9
1:2 63.2 60.8 1339.5 1288.4 104.0
1:4 33.5 30.4 710.9 644.2 110.4
1:8 16.0 15.2 340.1 322.1 105.6
1:16 8.0 7.6 169.8 161.1 105.4
4 Mean 106.3
— 146.0 146.0 3096.0 3096.0
1:2 74.0 73.0 1568.9 1548.0 101.3
1:4 36.8 36.5 780.3 774.0 100.8
1:8 18.9 18.3 400.1 387.0 103.4
1:16 9.8 9.1 207.8 193.5 107.4
5 Mean 103.2
— 166.0 166.0 3518.8 3518.8
1:2 88.1 83.0 1867.6 1759.4 106.1
1:4 43.9 41.5 930.9 879.7 105.8
1:8 21.4 20.7 452.7 439.8 102.9
1:16 11.0 10.4 233.5 219.9 106.2
6 Mean 105.3
— 152.3 152.3 3229.6 3229.6
1:2 76.3 76.2 1616.5 1614.8 100.1
1:4 41.2 38.1 873.0 807.4 108.1
1:8 20.1 19.0 427.1 403.7 105.8
1:16 10.2 9.5 215.5 201.9 106.8
Mean 105.2
Mean 104.6

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Spiking Recovery
Varying amounts of prolactin were added to six samples with endogenous prolactin levels of
4.1 to 15.2 ng/mL (86.9 to 322.2 μIU/mL). The recoveries ranged from 93.6% to 109.3% with
a mean of 102.4%.
Amount Added Observed Amount Added Observed
Sample (ng/mL) (ng/mL) (µIU/mL) (µIU/mL) Recovery %
1 — 4.1 — 86.9
51.7 58.9 1096.0 1248.7 105.9
104.1 112.3 2206.9 2380.8 103.9
154.0 163.1 3264.8 3457.7 103.2
Mean 104.3
2 — 6.5 — 137.8
51.7 63.0 1096.0 1335.6 109.3
104.1 113.0 2206.9 2395.6 102.3
154.0 161.2 3264.8 3417.4 100.5
Mean 104.0
3 — 5.0 — 104.9
51.7 60.9 1096.0 1291.1 108.2
104.1 114.4 2206.9 2425.3 105.1
154.0 160.2 3264.8 3396.2 100.8
Mean 104.7
4 — 14.9 — 315.9
38.6 56.2 818.3 1191.4 107.0
77.7 90.1 1647.2 1910.1 96.8
115.2 132.5 2442.2 2809.0 102.1
Mean 102.0
5 — 15.2 — 322.2
38.6 55.6 818.3 1178.7 104.7
77.7 87.9 1647.2 1863.5 93.6
115.2 124.8 2442.2 2645.8 95.1
Mean 97.8
6 — 10.2 — 216.2
38.6 51.1 818.3 1083.3 106.0
77.7 83.6 1647.2 1772.3 94.5
115.2 130.9 2442.2 2775.1 104.8
Mean 101.8
Mean 102.4

Spiking recovery testing was previously determined for the ADVIA Centaur Prolactin Assay.

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PRL ADVIA Centaur CP System

Precision
Precision was evaluated according to the CLSI protocol EP5-A2.11 According to this protocol,
the assay was tested daily, for 5 to 20 days, using multiple reagent lots, on multiple systems
across multiple sites. The system was calibrated immediately prior to the first test of day one.
Assay results were calculated using the two-point calibration. The following results were
obtained:
Within-run Between run Total
Mean (ng/mL) Mean (µIU/mL) CV(%) CV(%) CV(%)
4.4 93.2 3.4 4.8 5.9
10.8 229.0 3.5 5.0 6.1
25.7 544.8 3.2 5.4 6.3

Standardization
The ADVIA Centaur CP Prolactin assay standardization is traceable to World Health
Organization (WHO) 3rd IRP for human Prolactin (84/500). A comparison over the full assay
range gave the following correlation:
ADVIA Centaur CP PRL = 1.05 (WHO) – 0.4 IU/mL
r = 0.999
Assigned values for calibrators are traceable to this standardization.

Technical Assistance
For customer support, please contact your local technical support provider or distributor.
www.siemens.com/diagnostics

References
1. Ashby CD. Prolactin. In: Kaplan LA, Pesce AJ, editors. Methods in clinical chemistry. St. Louis: CV Mosby,
1987. p.258–65.
2. Vander AJ, Sherman JH, Luciano DS. Human physiology: the mechanisms of body function. New York:
McGraw-Hill Inc., 1985. p.589–91.
3. Liwnicz BH, Liwnicz RG. The hypothalamopituitary system. In: Kaplan LA, Pesce AJ, editors. Clinical
Chemistry: theory, analysis, and correlation. St. Louis: CV Mosby, 1989. p.613–9.
4. Butt WR, Blunt SM. The role of the laboratory in the investigation of infertility. Ann Clin Biochem
1988;25:601–9.
5. Albertsen PC, Chang TSK. Hormone measurements in the assessment of male infertility. J Clin Immunoassay
1983;6(1):51–6.
6. Owens O. Steroidal hormonal evaluation for common gynecological and testicular disorders. In: Kaplan LA,
Pesce AJ, editors. Methods in clinical chemistry. St. Louis: CV Mosby, 1987. p.216–7.
7. Clinical and Laboratory Standards Institute (formerly NCCLS). Procedures for the Handling and Processing
of Blood Specimens; Approved Guideline - Third Edition. Wayne, PA: Clinical and Laboratory Standards
Institute; 2004. NCCLS Document H18-A3.
8. Boscato LM, Stuart MC. Heterophilic antibodies: a problem for all immunoassays. Clin Chem 1988;34:27–33.
9. Clinical and Laboratory Standards Institute (formerly NCCLS). Interference Testing in Clinical Chemistry;
Approved Guideline - Second Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2005. NCCLS
document EP7-A2.
10. Clinical and Laboratory Standards Institute (formerly NCCLS). How to Define and Determine Reference
Intervals in the Clinical Laboratory; Approved Guideline - Second Edition. Wayne, PA: Clinical and
Laboratory Standards Institute; 2000. NCCLS Document C28-A2.
11. 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 EP5-A2.

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ADVIA Centaur, ReadyPack, and ACS:180 are trademarks of Siemens Healthcare Diagnostics.
© 2008 Siemens Healthcare Diagnostics Inc. All rights reserved.
US Pats 4,745,181; 4,918,192; 5,110,932; 5,609,822; 5,788,928

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PRL ADVIA Centaur CP System

Understanding the Symbols

The following symbols may appear on the product labeling:
Symbol Definition Symbol Definition

In vitro diagnostic medical device Catalog number

Authorized Representative in the European

Manufacturer
Community

CE Mark with identification number of notified

CE Mark
body

Consult instructions for use Caution! Potential Biohazard

Do not freeze (> 0°C) Temperature limitation (2–8°C)

Lower limit of temperature (≥ 2°C) Upper limit of temperature (≤ -10°C)

Keep away from sunlight Use by

Shake the reagent pack vigorously.

Refer to Loading Reagents in the assay-specific
Store upright ADVIA Centaur product instructions for detailed
information.

Batch code Contains sufficient for (n) tests

2010-01 Date format (year-month) Printed with soy ink

Green dot Recycle

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