Design Verification

liquicolor

1 Introduction .................................................................................................................................................................. 2
2 Imprecision .................................................................................................................................................................... 2
3 Linearity and Sensitivity............................................................................................................................................... 2
3.1 Linearity ................................................................................................................................................................................................. 2
3.2 Sensitivity .............................................................................................................................................................................................. 3
4 Comparison of Methods ............................................................................................................................................... 3
5 Stability Data ................................................................................................................................................................ 5
5.1 Accelerated Stress Tests .................................................................................................................................................................. 5
5.2 Real Time Stability.............................................................................................................................................................................. 5
6 Interference ................................................................................................................................................................... 7
7 Traceability .................................................................................................................................................................... 7

QM-Element: ISO9001:2000 No. 7.3.3 1/7

Rev. 006 | valid of 11.07.2012
1 Introduction
The performance characteristics of the ALBUMIN liquicolor hashave been tested and documented in order to verify
the clinical usefulness and compliance with the essential requirements of directive 98/79/EC.

2 Imprecision
The imprecision (within-run and day-to-day) of the ALBUMIN liquicolor method was calculated from six
determinations on ten consecutive days. Control sera with low, medium and high albumin levels were employed as
sample material.

Hitachi 717
Analyte concentration Intra-assay Inter-assay
(g/dl) SD (g/dl) %CV SD (g/dl) %CV
3.12 0.041 1.32 0.057 1.83
4.65 0.05 1.07 0.068 1.47
6.29 0.055 0.87 0.083 1.32

3 Linearity and Sensitivity

3.1 Linearity
The linearity of the ALBUMIN liquicolor method was controlled by employing dilutions of a high concentration pool
serum with physiological saline. The analysed concentrations were calculated vs. the regression line. Deviations
from the regression line are expressed in absolute and relative values.
As an example data measured on the HS600 are reported.

HS600
High Pool Analytical Data Regressed Data Deviation from Regression Line
Content (%) (g/dl) (g/dl) (g/dl) (%)
0 0.00 -0.02 0.02
10 0.84 0.80 0.04 4.55
20 1.68 1.62 0.06 3.49
30 2.07 2.44 -0.37 -15.3
40 3.17 3.26 -0.09 -2.85
50 4.19 4.08 0.11 2.63
60 5.14 4.90 0.24 4.84
70 5.89 5.72 0.17 2.93
80 6.69 6.54 0.14 2.18
90 7.31 7.36 -0.05 -0.71
100 7.94 8.18 -0.25 -3.02

Design Verification and Product Data for ALBUMIN liquicolor test 2/7
Rev. 006
Conclusion: ALBUMIN liquicolor is linear up to 6.5 g/dl.

3.2 Sensitivity
A 20-fold determination of a 0 calibrator (phys. saline) on a Hitachi 717 analyser revealed an absolute mean
of 0.07 g/dl with SD of 0.02 g/dl. Sensitivity was calculated on base of mean + 3 SD: 0.16 g/dl albumin.

4 Comparison of Methods
ALBUMIN liquicolor reagent lot # 0014 has been compared against a commercially available reagent method (Lot #
1198) measured on an Olympus AU400 analyzer. Control sera as well as patient samples have been employed in
the comparison (N=50). The results have been evaluated by a non-parametric regression analysis acc. to Passing &
Bablok. Acceptance criteria were r> 0.95 and a slope between 0.95 and 1.05. The linear regression obtained can be
described as follows:

r = 0.995
Y= 0.974 X + 0.260
Xmean = 4.5 g/dl
Ymean = 4.65 g/dl

Sample- Mean Reference Mean Test

no
1 4.76 4.90
2 4.69 4.86
3 4.89 5.03
4 4.62 4.69
5 3.89 4.04
6 3.24 3.50
7 4.56 4.76
8 4.96 5.07
9 3.08 3.41
10 3.08 3.27
11 5.25 5.42
12 4.80 4.96
13 5.24 5.33
14 4.54 4.57
15 4.32 4.41
16 4.78 4.86
17 4.73 4.88
18 4.58 4.80
19 4.88 4.99
20 4.73 4.79

Design Verification and Product Data for ALBUMIN liquicolor test 3/7
Rev. 006
 21 4.28 4.43
22 4.85 4.91
23 4.77 4.86
24 4.76 4.82
25 4.54 4.67
26 4.89 5.07
27 4.79 4.97
28 4.01 4.04
29 4.55 4.66
30 4.82 4.89
31 4.54 4.83
32 4.55 4.70
33 3.61 3.80
34 4.94 5.05
35 5.02 5.16
36 3.88 4.02
37 5.33 5.57
38 4.36 4.55
39 5.28 5.27
40 4.57 4.71
41 4.89 5.13
42 4.90 5.05
43 4.83 5.00
44 3.18 3.32
45 4.73 4.97
46 4.71 4.73
47 4.69 4.87
48 1.96 2.09
49 4.88 5.07
50 4.61 4.72

Conclusion: The methods showed a very good agreement and no significant deviation could be observed.

Design Verification and Product Data for ALBUMIN liquicolor test 4/7
Rev. 006
5 Stability Data
5.1 Accelerated Stress Tests
Recovery in Control Sera
Commercially available control sera have been employed. The control sera have been reconstituted/prepared
according to the manufacturer’s instructions. Fivefold determinations of each control serum have been performed
with ALBUMIN liquicolor method after 10 and 12 days storage at 56°C. The means of the fivefold determinations
have been calculated and compared with the values of fresh reagent.

CONTROL SERUM ALBUMIN Reagent ALBUMIN Reagent 10 d 56°C ALBUMIN Reagent 12 d 56°C
RECOVERY fresh
Control LOT Result Result Deviation Result Deviation
serum g/dl g/dl (%) g/dl (%)
HumaTrol N # 018 3.45 3.29 -4.64 3.34 -3.19
HumaTrol P # 016 4.72 4.56 -3.39 4.63 -1.91
SERODOS # 6868 4.67 4.53 -3 4.54 -2.78
SERO.plus # 6795 3.75 3.61 -3.73 3.61 -3.73
Precinorm # 150092 3.60 3.44 -4.44 3.51 -2.50
Precipath # 199459 3.41 3.26 -4.40 3.32 -2.64
Mean 3.93 3.78 -3.82 3.83 -2.54

Linearity

The linearity of ALBUMIN liquicolor was controlled according to the procedure already described in 2.1. The
reagents were stressed 10 and 12 days at 56°C.

ALBUMIN reagent 10 days 56°C ALBUMIN reagent 12 days 56°C

High Pool Analytical Regressed Deviation from Regression Analytical Regressed Deviation from Regression
Data Data Line Data Data Line
Content (%) g/dl g/dl g/dl (%) g/dl g/dl g/dl (%)
0 0.01 -0.04 0.05 -112.24 -0.01 0.02 -0.02 -127
10 0.66 0.84 -0.18 -21.92 0.66 0.87 -0.21 -24.3
20 1.68 1.72 -0.04 -2.24 1.69 1.73 -0.04 -2.35
30 2.61 2.60 0.01 0.45 2.70 2.58 0.12 4.50
40 3.74 3.48 0.26 7.39 3.62 3.43 0.18 5.33
50 4.44 4.36 0.08 1.89 4.46 4.29 0.17 4.07
60 5.23 5.24 -0.01 -0.14 5.25 5.14 0.11 2.06
70 5.95 6.12 -0.17 -2.73 5.87 5.99 -0.13 -2.12
80 6.88 7.00 -0.12 -1.74 6.67 6.85 -0.18 -2.56
90 7.71 7.88 -0.17 -2.18 7.49 7.70 -0.21 -2.71
100 8.38 8.76 -0.38 -4.30 8.39 8.55 -0.16 -1.90

5.2 Real Time Stability

The stability of ALBUMIN liquicolor has been tested on real-time storage conditions over a period of 24, 27 and 28
months for three independent production lots (H022, exp. date: Nov. 2004; H013, exp. date: Nov. 2003; H020, exp.
date: Jun. 2004), measurements were made on Hitachi 717. Acceptance criteria were based on the recovery of
controls and linearity.

Recovery of Control Sera

Commercially available control sera have been employed. The control sera have been reconstituted/prepared
according to the manufacturer’s instructions. Twofold determinations of each control serum have been performed
with the ALBUMIN liquicolor test method. The means of the twofold determinations have been calculated and
compared with the values of the reference reagent.

Design Verification and Product Data for ALBUMIN liquicolor test 5/7
Rev. 006
 CONTROL Reference Lot. H022 24 months Lot. H013 27 months Lot. H020 28 months
Reagent
H032
Control LOT Result Result Deviation Result Deviation Result Deviation
serum g/dl g/dl (%) g/dl (%) g/dl (%)

HumaTrol N # 019 2.44 2.6 6.55 2.54 4.1 2.58 5.74

HumaTrol N # 020 2.59 2.7 4.25 2.68 3.47
HumaTrol P # 016 4.72 4.75 0.63 3.42 -27.5 3.55 -24.8
HumaTrol P # 017 3.42 3.62 5.85 3.46 1.17 3.49 2.04
SERODOS # 6869 3.63 3.69 1.65 3.56 -1.93
SERODOS # 6868 3.53 3.53 0
SERO.plus # 6796 2.71 2.77 2.21 2.75 1.48 2.74 1.11
Precinorm # 164919 3.52 3.72 5.68 3.59 1.99
Precinorm # 159296 2.54 2.58 1.57
Precipath # 163661 2.40 2.47 2.92 2.43 1.25
Precipath # 165215 2.51 2.52 0.40

Linearity
ALBUMIN liquicolor Lot H032, Reference ALBUMIN liquicolor Lot H022, 24 months
High Pool Analytical Regressed Deviation from Regression Analytical Regressed Deviation from Regression
Data Data Line Data Data Line
Content (%) g/dl g/dl g/dl (%) g/dl g/dl g/dl (%)
0 0.00 0.00 0.00 -100.00 0.85 12.45 -11.60 -93.17
10 0.71 0.78 -0.08 -10.00 234 219 15.17 6.92
20 1.51 1.57 -0.06 -3.82 433 426 6.59 1.55
30 2.42 2.36 0.06 2.69 621 633 -12.29 -1.94
40 3.26 3.14 0.12 3.71 842 840 2.13 0.25
50 4.00 3.93 0.07 1.65 1012 1047 -35.20 -3.36
60 4.69 4.72 -0.03 -0.67 1250 1254 -3.73 -0.30
70 5.47 5.50 -0.03 -0.61 1421 1461 -39.11 -2.68
80 6.25 6.29 -0.04 -0.72 1594 1667 -72.99 -4.38
90 7.01 7.08 -0.07 -0.94 1768 1874 -106.37 -5.67
100 7.58 7.86 -0.29 -3.67 1997 2081 -84.25 -4.05

ALBUMIN liquicolor Lot H013, 27 months ALBUMIN liquicolor Lot H020, 28 months
High Pool Analytical Regressed Deviation from Regression Analytical Regressed Deviation from Regression
Data Data Line Data Data Line
Content (%) g/dl g/dl g/dl (%) g/dl g/dl g/dl (%)
0 -0.60 15.53 -16.13 -103.86 -0.65 20.30 -20.95 -103.20
10 260 254 5.28 2.08 257 256 0.72 0.28
20 490 493 -3.41 -0.69 489 491 -2.56 -0.52
30 751 732 18.55 2.53 760 727 32.55 4.48
40 974 971 3.26 0.34 964 963 1.32 0.14
50 1225 1210 14.52 1.20 1221 1198 22.54 1.88
60 1434 1449 -14.77 -1.02 1410 1434 -24.15 -1.68
70 1680 1688 -7.31 -0.43 1660 1669 -9.48 -0.57
80 1913 1927 -13.30 -0.69 1868 1905 -37.16 -1.95
90 2142 2166 -23.59 -1.09 2072 2141 -68.80 -3.21
100 2293 2404 -111.08 -4.62 2244 2376 -132.08 -5.56

Conclusion: All results support the claimed stability of 24 months from production for the test kit.

Design Verification and Product Data for ALBUMIN liquicolor test 6/7
Rev. 006
6 Interference
Interferences by ascorbate, bilirubin, hemoglobin, triglycerides and lipemia have been studied by adding known
amounts of the potentially interfering substance to a known sample.
Recoveries have been analyzed according to the method of Glick et al. (Clin.Chem. 1986, 32 470-5).

The results are summarised in the following table.

Ascorbate Result Dev. Bilirubin Result Dev. Hemoglobin Result Dev.

mg/dl g/dl % mg/dl g/dl % mg/dl g/dl %
0 3.83 100 0 3.78 100 0 3.80 100
2 3.86 100.8 4 3.67 97.2 50 3.95 103.9
4 3.88 101.3 8 3.69 97.7 100 3.90 102.6
6 3.92 102.5 12 3.52 93.1 150 3.90 102.6
8 3.83 100.1 16 3.58 94.8 200 3.85 101.3
10 3.82 99.9 20 3.51 92.8 250 3.90 102.6
12 3.81 99.6 24 3.43 90.9 300 3.90 102.6
14 3.85 100.7 28 3.38 89.4 350 4.00 105.3
16 3.93 102.7 32 3.30 87.3 400 3.90 102.6
18 3.82 99.7 36 3.24 85.8 450 3.95 103.9
20 3.84 100.3 40 3.24 85.8 500 3.95 103.9

Glick 1 Glick 2 2

Triglycerid. Result Dev. Intralipid Result Dev.

mg/dl g/dl % mg/dl g/dl %
0 3.79 100 0 3.75 100
250 3.89 102.6 100 3.64 97.2
500 3.91 103.2 200 3.53 94.1
750 4.02 106.1 300 3.52 94.0
1000 3.95 104.2 400 3.45 92.1
1250 3.91 103.3 500 3.43 91.6
1500 3.99 105.3 600 3.40 90.8
1750 3.94 104.0 700 3.30 88.0
2000 4.08 107.8 800 3.24 86.4
2250 1.01 105.9 900 3.15 84.0
2500 3.99 105.4 1000 3.02 80.5

Glick 2 2

Conclusion: This test is not or only slightly influenced by ascorbate, bilirubin, haemoglobin, triglycerides and
intralipid (lipemia).

7 Traceability
ALBUMIN liquicolor is calibrated with either a standard supplied with the kit or AUTOCAL, both are traceable to the
reference material CRM 470.

Design Verification and Product Data for ALBUMIN liquicolor test 7/7
Rev. 006