Design Verification

HM Hb P

C

1 Imprecision................................................................................................................................................................2
1.1 Within-run Imprecision............................................................................................................................................2
1.2 Day-to-day Imprecision ...........................................................................................................................................2
2 Linearity ....................................................................................................................................................................3
2.1 Linearity ......................................................................................................................................................................3
3 Comparison of Methods .........................................................................................................................................4
3.1 Study 1 ........................................................................................................................................................................4
3.2 Study 2 ........................................................................................................................................................................4
3.3 Study 3 ........................................................................................................................................................................5
4 Comparison of Two Instruments.............................................................................................................................5
5 Stability and Storage ................................................................................................................................................6
5.1 Real Time Stability Reagent ..................................................................................................................................6
5.2 Real Time Stability Calibrators.............................................................................................................................7
5.3 Real Time Stability Controls..................................................................................................................................8
6 Transport Simulation................................................................................................................................................8

QM-Element: ISO9001:2000 No. 7.3.3 1/8

Rev. 004 | valid of 29.05.2008
1 Imprecision

1.1 Within-run Imprecision

The imprecision of the HumaMeter Hb Plus System, consisting of the HumaMeter Hb Plus Photometer and
HumaMeter Hb Plus Reagent prefilled cuvettes was calculated from 20 routine samples which were determined at
the same day in 6 consecutive measurements. The data were obtained from one instrument and one reagent lot.
The imprecision is monitored as coefficient of variation (CV%) in the below listed table.
Within run measurements
Sample 1 2 3 4 5 6 ∅ CV%

1 14,3 14,4 14,7 14,4 14,7 14,5 14,5 1%

2 13,3 13,0 12,8 13,1 12,7 13,0 13,0 2%

3 10,7 10,5 10,3 10,3 10,5 10,7 10,5 2%

4 13,0 13,2 13,3 13,4 13,6 13,8 13,4 2%

5 12,3 12,1 12,4 12,1 12,1 12,2 12,2 1%

6 9,0 9,4 9,4 9,4 9,1 8,8 9,2 3%

7 13,3 13,6 13,7 14,0 14,2 14,3 13,9 3%

8 8,5 8,6 8,8 8,9 8,7 9,1 8,8 2%

9 13,4 13,5 13,9 13,6 13,2 13,5 13,5 2%

10 8,4 8,2 8,0 7,8 8,1 8,0 8,1 3%

11 11,1 10,9 10,8 10,6 10,9 11,2 10,9 2%

12 15,3 15,1 15,0 15,2 15,2 15,4 15,2 1%

13 13,4 13,0 12,9 12,5 12,5 12,8 12,9 3%

14 12,0 12,1 11,9 11,9 11,8 11,5 11,9 2%

15 9,1 9,0 9,1 9,4 9,4 9,6 9,3 3%

16 10,7 10,4 10,1 10,2 10,5 10,6 10,4 2%

17 12,2 11,9 11,5 11,4 11,6 11,9 11,8 3%

18 11,3 11,5 11,1 11,0 11,0 10,6 11,1 3%

19 13,2 13,1 13,1 13,3 13,2 13,0 13,2 1%

20 13,4 13,7 13,4 13,1 12,9 12,6 13,2 3%

1.2 Day-to-day Imprecision

The day-to-day imprecision was based on the measurement of 6 AHD calibrators on 10 consecutive days The
imprecision is expressed as coefficient of variation in below listed table.
Calibrators

Day 3 g/dl 6 g/dl 9 g/dl 12 g/dl 15 g/dl 18 g/dl

1 3,1 6,0 9,2 11,9 15,2 18,1

2 3,0 6,2 9,1 11,9 14,8 17,7

3 2,9 6,0 9,3 12,0 14,9 18,2

4 2,8 6,0 8,7 11,8 15,3 18,3

5 2,9 5,9 9,4 11,9 15,0 18,2

6 2,8 6,1 9,4 12,0 15,0 18,1

7 2,9 6,2 9,3 11,9 15,2 18,3

8 3,0 6,4 9,5 12,3 15,0 17,6

9 3,0 5,8 8,8 11,8 14,8 18,3

10 2,9 6,4 9,4 12,2 14,9 18,1

∅ 2,9 6,1 9,2 12,0 15,0 18,1

CV% 3% 3% 3% 1% 1% 1%

Conclusion:
The above results confirm the low imprecision and the good reproducibility of the system.

Design Verification and Product Data for HumaMeter Hb Plus 2/8

Rev. 004
2 Linearity

2.1 Linearity
To determine the measuring range, a dilution series was produced from 10 routine blood samples mixed 1:1 with
calcium-free phosphate buffer. The hemoglobin concentration was then determined for each individual stage of
the dilution series. Each dilution series comprised 5 dilution stage. Next, the stage with the highest degree of
dilution was mixed again 1:1 with the next highest dilution stage.

A= Measured values Hb (g/dl)

B= Expected values Hb ( g/dl)

18

y = 1,0033x + 0,0544
16
R2 = 0,9968
14

12
A:
10

0
0 2 4 6 8 10 12 14 16 18

B:

Design Verification and Product Data for HumaMeter Hb Plus 3/8

Rev. 004
3 Comparison of Methods
The HumaMeter Hb Plus system was compared against the Coulter AC T8 ( SLS Method ), 80 routine samples have
been employed in this study and against the reference method HEMOGLOBIN liquicolor . The results are shown in
the following diagrams.
3.1 Study 1
HumaMeter Hb Plus versus SLS Method
Comparison with autmated SLS-Method Hb ( g/dl )
B: HumaMeter Hb Plus A: SLS
1 8,0

y = 1,0107x - 0,1031
1 6,0
R2 = 0,9843
1 4,0

1 2,0

B 1 0,0
8 ,0

6 ,0

4 ,0

2 ,0

0 ,0
0 ,0 2,0 4 ,0 6 ,0 8,0 1 0,0 12 ,0 1 4,0 1 6,0

3.2 Study 2
HumaMeter Hb Plus versus H liquicolor

Comparison with Hemoglobin Liquicolor A: HumaMeter Hb Plus B: Hemoglobin Liquicolor

20
19
18
17
y = 0,9401x +
16
15 R = 0,776
14

A13
12
11
10

9
8
7
6

5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
B

Design Verification and Product Data for HumaMeter Hb Plus 4/8

Rev. 004
 3.3 Study 3
HumaMeter Hb Plus versus H

20
HumaMeter Hb plus comparison with reference method : Hemoglobin Liquicolor
19
18 Hemoglobin Liquicolor Cyan Methemoglobin Method [g/dl] A
respective Method [g/dl] B
17
16 y = 0,8759x + 1,4684
15 R² = 0,9083

14
13
12
y = 0,8138x + 0,4624
B 11 R2 = 0,8968
10
9
hemocue
8
Hbplus rgt 1,5 ml
7
6 Linear (hem ocue)

5
4
3
2
1
0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

A
Conclusion:
Compared against the reference method cyanmethhemoglobin the Hemocue exhibits a significant
bias in the physiological relevant concentration range. HumaMeter Hb Plus, however, correlates well
in this concentration range.

4 Comparison of Two Instruments

21

y = 0,9705x + 0,5074
R2 = 0,995
19
Measured concentrations [g/dl] Hbplus 2

17

15

13

11

9
9 11 13 15 17 19 21
Measured concentration [g/dl] Hbplus 1

Design Verification and Product Data for HumaMeter Hb Plus 5/8

Rev. 004
Conclusion:

The data show an excellent correlation between two instruments.

5 Stability and Storage
The stability of the HumaMeter Hb Plus reagent, calibrators and controls have been evaluated by real time studies
over a storage period of 29 months.
Reagent, calibrators and controls have been stored at ambient conditions.
5.1 Real Time Stability Reagent
At suitable intervals function tests have performed with 6 different calibrators in concentrations of 3, 6, 9, 12, 15
and 18 g/dl Hb. Differences from target concentrations are expressed in % . The results are summarised below.
Calibrator recoveries
Months 3 g/dl 6 g/dl 9 g/dl 12 g/dl 15 g/dl 18 g/dl
3 0,1 0 -0,35 0,345 0,14 -0,42
6 -0,07 0,21 0,31 -0,1 -0,59 0
12 -0,03 -0,03 -0,14 0,14 -0,03 -0,91
18 0,07 -0,14 -0,35 0,21 -0,03 0,45
24 -0,1 0 -0,17 0,24 0,24 0,56
29 -0,03 0 -0,42 0,1 -0,35 -0,45

Real time stability

8
6
% Dev. from target

3 g/dl
4
6 g/dl
2
9 g/dl
0
12 g/dl
-2 3 6 12 18 24 29
15 g/dl
-4
18 g/dl
-6
-8
Storage time, Months

Conclusion:

The storage life tests showed that all of the reagents tested were very stable and suffered no loss of performance
even after a longer period of storage. This fact, combined with the basic chemical characteristics of the substances
tested (Triton-X, NaOH and Hämin) allows the stability of the reagents to be guaranteed for 36 months. The
transport simulation revealed no weaknesses in the reagent vials or boxes. Here, too, no negative influence on the
quality of the products is to be expected.

Design Verification and Product Data for HumaMeter Hb Plus 6/8

Rev. 004
5.2 Real Time Stability Calibrators
Expected values:
Calibrators
Hb- 3 g/dl 6 g/dl 9 g/dl 12 g/dl 15 g/dl 18 g/dl
concentration
Target value 0.086 0.172 0.257 0.343 0.429 0.515
Measurement 0.081 – 0.091 0.162 – 0.182 0.242 – 0.272 0.322 – 0.364 0.403 – 0.455 0.484 – 0.546
range

Measured values:
Lot 1
3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
3 0.089 3.11 0.084 2.94 0.085 2.97 0.088 3.08 0.083 2.90 0.085 2.97 0.087 3.04
6 0.172 6.01 0.178 6.22 0.171 5.98 0.168 5.87 0.172 6.01 0.172 6.01 0.174 6.08
9 0.247 8.64 0.266 9.30 0.253 8.84 0.247 8.64 0.252 8.81 0.245 8.57 0.253 8.84
12 0.353 12.34 0.340 11.89 0.347 12.13 0.349 12.20 0.350 12.24 0.346 12.10 0.340 11.89
15 0.433 15.14 0.412 14.40 0.428 14.96 0.428 14.96 0.436 15.24 0.419 14.65 0.421 14.72
18 0.503 17.58 0.515 18.00 0.489 17.10 0.528 18.46 0.531 18.56 0.502 17.55 0.521 18.21

Lot 2
3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
3 0.088 3.08 0.083 2.90 0.084 2.97 0.085 2.97 0.085 2.97 0.087 3.04 0.086 3.01
6 0.169 5.91 0.180 6.29 0.169 5.98 0.179 6.26 0.165 5.77 0.165 5.77 0.168 5.87
9 0.249 8.71 0.261 9.12 0.247 8.84 0.246 8.60 0.262 9.16 0.262 9.16 0.254 8.88
12 0.334 11.68 0.342 11.96 0.337 12.13 0.339 11.85 0.340 11.89 0.349 12.20 0.337 11.78
15 0.442 15.45 0.441 15.42 0.437 14.96 0.428 14.96 0.431 15.07 0.430 15.03 0.434 15.17
18 0.507 17.72 0.497 17.38 0.497 17.10 0.524 18.32 0.519 18.14 0.533 18.63 0.523 18.28

Lot 3
3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
3 0.085 2.97 0.090 3.15 0.089 3.11 0.090 3.15 0.084 2.94 0.089 3.11 0.088 3.08
6 0.171 5.98 0.179 6.26 0.173 6.05 0.175 6.12 0.174 6.08 0.169 5.91 0.175 6.12
9 0.264 9.23 0.264 9.23 0.253 8.84 0.263 9.19 0.250 8.74 0.264 9.23 0.263 9.19
12 0.335 11.71 0.341 11.92 0.352 12.31 0.351 12.27 0.349 12.20 0.350 12.24 0.348 12.17
15 0.427 14.93 0.427 14.93 0.445 15.56 0.420 14.68 0.418 14.61 0.435 15.21 0.425 14.86
18 0.516 18.04 0.494 17.27 0.499 17.45 0.500 17.48 0.498 17.41 0.518 18.11 0.523 18.28

Conclusion:

The real time stability tests of the calibrators showed that is no loss of performance even after storage for 39
months.

Design Verification and Product Data for HumaMeter Hb Plus 7/8

Rev. 004
5.3 Real Time Stability Controls

Lot 1
fresh 3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
6 0.086 6.01 0.169 5.91 0.180 6.29 0.169 5.91 0.179 6.26 0.165 5.77 0.165 5.77 0.168 5.87
15 0.429 15.00 0.442 15.45 0.441 15.42 0.437 15.28 0.428 14.96 0.431 15.07 0.430 15.03 0.434 15.17

Lot 2
fresh 3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
6 0.086 6.01 0.171 5.98 0.179 6.26 0.173 6.05 0.175 6.12 0.174 6.08 0.169 5.91 0.175 6.12
15 0.429 15.00 0.427 14.93 0.427 14.93 0.445 15.56 0.420 14.68 0.418 14.61 0.435 15.21 0.425 14.86

Lot 3
fresh 3 months 6 months 12 months 18 months 24 months 29 months 39 months
Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb Abs. Hb
g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl g/dl
6 0.086 6.01 0.172 6.01 0.178 6.22 0.171 5.98 0.168 5.87 0.172 6.01 0.172 6.01 0.174 6.08
15 0.429 15.00 0.433 15.14 0.412 14.40 0.428 14.96 0.428 14.96 0.436 15.24 0.419 14.65 0.421 14.72

Conclusion:

The real time stability tests of the calibrators showed that is no loss of performance even after storage for 39
months.

6 Transport Simulation
To determine the suitability and robustness of the reagent vials and boxes, 2 of each of the 3 types of vials (with
the outer box) were placed in an orbital shaker with one vial of each type inserted upside down. A pH indicator
strip was inserted into each box to show any reagent leakage. Additionally, the boxes with vials were subjected to
a 1-meter fall to the laboratory floor once every day. At the conclusion of the 3-week test period all of the reagent
vials and their boxes were undamaged. None of the pH strips showed any evidence of leakage of reagents or
standard.

Design Verification and Product Data for HumaMeter Hb Plus 8/8

Rev. 004