Design Verification

liquicolor

1 Introduction .................................................................................................................................................................. 2
2 Imprecision .................................................................................................................................................................... 2
3 Linearity and Detection Limit ...................................................................................................................................... 2
Linearity Tested with Standards .......................................................................................................................................................... 2
Detection Limit ........................................................................................................................................................................................... 3
4 Tracebility ...................................................................................................................................................................... 4
5 Recovery of Control Sera .............................................................................................................................................. 4
6 Comparison of Methods ............................................................................................................................................... 5
7 Interferences ................................................................................................................................................................. 5
8 Stability.......................................................................................................................................................................... 6
Recovery in Control Sera ......................................................................................................................................................................... 6
Linearity......................................................................................................................................................................................................... 7

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

Rev. 006 | valid of 05.12.2016
1 Introduction
The imprecision and the recovery of the GLUCOSE liquicolor test has been tested and documented in order to verify
its clinical usefulness and additional features such as linearity, interferences and method comparison.

2 Imprecision
The imprecision (within-run and day-to-day) of the GLUCOSE liquicolor method was calculated from six
determinations on six consecutive days. Three pooled control sera were employed as sample material.

Analyser: HITACHI 717

Analyte concentration Intra-assay Inter-assay
(mg/dl) SD (mg/dl) %CV SD (mg/dl) %CV
51.0 0.886 1.74 1.095 2.15
117.15 3.508 2.99 3.546 3.03
379.89 14.325 3.77 14.789 3.89

3 Linearity and Detection Limit

Linearity Tested with Standards
The linearity of the GLUCOSE liquicolor method was controlled by employing dilutions of a patient serum pool
which has been spiked with glucose to approx. 500 mg/dl. The analysed concentrations were calculated vs. the
regression line. The deviation between analytical and regressed data are expressed in absolute and relative values.
As an example data measured on the Hitachi 717 are reported.

High Pool Analytical Data Regressed Data Deviation from Regression Line
Content (%) mg/dl mg/dl mg/dl (%)
0 0.05 -0.13 0.18 -138.89
10 51.35 52.01 -0.66 -1.27
20 104.65 104.15 0.50 0.48
30 156.40 156.30 0.10 0.07
40 208.50 208.44 0.06 0.03
50 260.40 260.58 -0.18 -0.07
60 310.30 312.72 -2.42 -0.77
70 361.70 364.86 -3.16 -0.87
80 412.15 417.00 -4.85 -1.16
90 441.90 469.14 -27.24 -5.81
100 484.20 521.29 -37.09 -7.11

Design Verification and Product Data for GLUCOSE LIQUICOLOR 2/7

Rev. 006
 600
L IN E A R IT Y

M E A S U R E D A N A L Y T E C O N C E N T R A T IO N ( m g /d l)
Er r o r B a r s = ± 2 S D
500

400

300

200

100

0
0 20 40 60 80 100 120
HIG H P O O L C O NT ENT ( % )

Conclusion: The GLUCOSE liquicolor test is linear up to a glucose concentration of at least 400 mg/dl or
22.2 mmol/l.

Detection Limit
A 20-fold determination of a ‘0’ sample (phys. saline) on a HumaStar 100 analyser revealed an absolute mean of
2.32mg/dl and a SD of 0.42mg/dl. From the three-fold standard deviation the detection limit can therefore be
calculated on the base of mean +3 SD to 3.58mg/dl.

Design Verification and Product Data for GLUCOSE LIQUICOLOR 3/7

Rev. 006
4 Tracebility
Three levels of the international reference preparation SRM 965 has been measured in the GLUCOSE liquicolor test
in 5-fold determinations with two different reagent lots:

Target Confid. range Lot A Lot B

SRM 965 Level 2 199.93 196.39 – 203.47 199.1 199.7
196.3 203.1
199.4 202.6
198.9 198.2
198.6 202.7
Mean 198.46 201.26

SRM 965 Level 3 294.65 291.33 – 297.97 289.9 297.3

293.7 292.7
290.6 301.0
298.4 298.1
296.2 292.3
Mean 293.76 296.28

As can be seen, the recovery of the reference preparation is excellent when calibrated with the respective kit
standards.

Conclusion: The method is traceable to the reference preparation SRM 965.

5 Recovery of Control Sera

A number of commercially available control sera have been employed. The control sera have been
reconstituted/prepared according to the manufacturer’s instructions. Five-fold determinations of each control
serum have been performed with GLUCOSE liquicolor reagents of different production lots. The mean values have
been calculated and compared with the target values of the respective control sera.

CONTROL SERUM RECOVERY Fresh Reference 10 days 37°C 2 weeks 25°C

Control LOT Target Range Result Deviation Result Deviation Result Deviation Result Deviation
serum mg/dl mg/dl mg/dl (%) mg/dl (%) mg/dl (%) mg/dl (%)

HUMATROL N # 018 122 103-140 114.8 -5.89 118.2 -3.13 114.7 -6.00 117.2 -3.93
HUMATROL P # 016 254 215-292 232.4 -8.49 241.6 -4.88 237.4 -6.54 236.6 -6.86
SERODOS # 6868 115.6 98.3-133 119.0 2.91 122.2 5.67 118.7 2.72 119.7 3.55
SERO.Plus # 6795 248.7 211.4-286 255.7 2.82 263.7 6.02 257.0 3.32 260.0 4.54
Precinorm # 154916 98 83.3-112.7 103.5 5.61 106.1 8.24 104.0 6.12 104.9 7.02
Precipath # 199459 235 199-271 248.0 5.54 253.5 7.86 249.8 6.28 251.2 6.89
Summary 1073.3 1073.5 1105.2 1081.5 1089.5

Conclusion: No significant differences could be observed between fresh and stressed reagents, confirming the
stability of the reagents. Controls were all recoverd well within the specified ranges.

Design Verification and Product Data for GLUCOSE LIQUICOLOR 4/7

Rev. 006
6 Comparison of Methods
The GLUCOSE liquicolor test has been compared against a commercially available glucose method. Control sera as
well as patient samples have been employed in the comparison.
The results have been evaluated by a main component analysis. The linear regression obtained could be described
as follows:

N= 55
r= 0.997
Y= 1.000 * X + 2.788
Xmean = 100,89
Ymean = 103,44

Conclusion: Both methods showed a good agreement and no significant deviation could be observed with any
specific sample.

7 Interferences
Interference by uric acid, glutathion, anticoagulants, bilirubin and creatinine 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.Cem. 1986, 32 470-5).

The results are summarised in the following table.

Uric Acid Analyt. Dev. Ascorb. Analyt. Dev. Glutathion Analyt. Dev. Citrate Analyt. Dev.
mg/dl Result % mg/dl Result % mg/dl Result % mg/ml Result %
mg/dl mg/dl mg/dl mg/dl
0 129.5 100.0 0 115.2 100.0 0 99.0 100.0 0 102.6 100.0
1.4 128.8 99.5 2 111.6 96.9 5 95.3 96.3 2.5 102.7 100.1
2.8 129.3 99.8 4 108.2 94.0 10 93.4 94.3 5.0 100.5 98.0
4.2 129.0 99.6 6 107.4 93.2 15 90.8 91.7 7.5 102.4 99.8
5.6 129.4 99.9 8 104.8 91.0 20 89.0 89.9 10.0 101.1 98.5
7.0 128.9 99.5 10 104.8 91.0 25 86.0 86.9 12.5 102.3 99.7
8.4 130.2 100.5 12 102.5 89.0 30 84.7 85.5 15.0 101.9 99.4
9.8 131.6 101.6 14 99.1 86.1 35 83.0 83.8 17.5 101.6 99.1
11.2 130.2 100.5 16 96.4 83.7 40 81.4 82.2 20.0 103.2 100.6
12.6 131.5 101.5 18 94.1 81.7 45 79.8 80.6 22.5 102.8 100.2
14.0 132.4 102.2 20 92.8 80.6 50 79.9 80.7 25.0 103.8 101.2

Glick 1 2 2 1

Design Verification and Product Data for GLUCOSE LIQUICOLOR 5/7

Rev. 006
 Heparin Analyt. Dev. EDTA Analyt. Dev. Bilirubin Analyt. Dev. Creatinin Analyt. Dev.
U/ml Result % mg/ml Result % mg/dl Result % e mg/dl Result %
mg/dl mg/dl mg/dl mg/dl
0 106.5 100.0 0 99.7 100.0 0 112.3 100.0 0 130.4 100.0
2.5 107.0 100.5 0.5 97.7 98.0 4 107.9 96.1 1.5 132.4 101.5
5.0 108.2 101.6 1.0 96.1 96.4 8 105.9 94.3 3.0 130.1 99.7
7.5 107.1 100.6 1.5 97.8 98.1 12 100.8 89.7 4.5 129.9 99.6
10.0 107.7 101.2 2.0 99.1 99.4 16 100.0 89.0 6.0 131.0 100.5
12.5 107.2 100.7 2.5 97.3 97.6 20 97.4 86.7 7.5 129.8 99.5
15.0 105.5 99.1 3.0 99.4 99.7 24 93.8 83.5 9.0 128.6 98.6
17.5 106.2 99.8 3.5 98.2 98.5 28 91.2 81.2 10.5 129.5 99.3
20.0 106.5 100.0 4.0 98.8 99.1 32 90.1 80.2 12.0 133.1 102.1
22.5 107.2 100.7 4.5 98.3 98.6 26 87.5 77.9 13.5 130.2 99.8
25.0 106.7 100.2 5.0 98.9 99.2 40 86.7 77.2 15.0 128.7 98.7

Glick 1 2 3 1

Conclusion: This test is not influenced by uric acid, ascorbic acid, glutathion, anticoagulants and creatinine in
physiological concentrations. A Glick value of 3 for bilirubin indicates an interfering effect. Samples with highly
pathological bilirubin concentrations (> 12 mg/dl) should therefore be avoided.

8 Stability
Recovery in Control Sera
Commercially available control sera have been employed. The control sera have been reconstituted/prepared
according to the manufacturer’s instructions. Five-fold determinations of each control serum have been performed
with the GLUCOSE liquicolor test method. The means of the fivefold determinations have been calculated and
compared with the target values.

CONTROL SERUM RECOVERY Reference 24 months 26 months 27 months

Control LOT Target Range Result Deviation Result Deviation Result Deviation Result Deviation
serum mg/dl mg/dl mg/dl (%) mg/dl (%) mg/dl (%) mg/dl (%)

HUMATROL N # 018 114 97-131 109.8 1.04 113.0 1.01 111.5 1.02 111.1 1.02
HUMATROL N # 019 123 105-141.5 113.5 1.08 115.7 1.06 114.9 1.07 114.2 1.08
HUMATROL P # 016 234 199-269 214.7 1.09 218.3 1.07 218.0 1.07 215.6 1.08
HUMATROL P # 017 250.7 213.1- 232.6 1.08 238.1 1.05 235.0 1.07 233.9 1.07
288.3
SERODOS # 6868 115.6 98.3-133 111.0 1.04 113.3 1.02 110.8 1.04 110.6 1.04
SERO.Plus # 6795 248.7 211.4-286 243.5 1.02 246.0 1.01 242.0 1.03 242.6 1.03
SERO.Plus # 6796 274 233-315 241.6 1.13 243.4 1.12 241.6 1.13 239.2 1.14
Precinorm #159296 102 87-117 101.1 1.01 102.5 1.00 101.0 1.01 101.3 1.01
Precipath #165215 242 206-278 239.3 1.01 244.4 0.99 241.0 1.00 211.2 1.17
Summary 1704 1607.1 1634.7 1615.8 1579.7

Conclusion: The above results clearly demonstrate the stability of the test for more than 24 months after
production. In addition to the real-time stability studies, temperature stress tests are routinely performed on each
production batch as part of the end control of the product.

Design Verification and Product Data for GLUCOSE LIQUICOLOR 6/7

Rev. 006
 Linearity
Four independent lots have been stored under the recommended conditions for up to 27 months. A high
concentration pool serum was employed and dilutions with phys. saline were made. The analysed concentrations
were calculated vs. the regression line. Deviation from the regression line are expressed in absolute and relative
values.

24 months 25 months 26 months 27 months

High pool Analytical Data Regression Analytical Data Regression Analytical Data Regression Analytical Data Regression
Content (%) mg/dl Line (%) mg/dl Line (%) mg/dl Line (%) mg/dl Line (%)
0 -0.10 -109 0.25 -76.0 0.15 -86.96 0.25 -80.35
10 47.35 4.84 46.75 3.34 46.35 4.75 47.00 6.14
20 89.05 -0.19 89.55 0.12 87.50 0.18 87.10 -0.22
30 131.9 -1.03 132.9 -0.55 130.3 -0.11 128.6 -1.27
40 177.9 0.32 178.6 0.43 172.1 -0.26 173.1 -0.12
50 216.9 -2.00 219.6 -1.07 213.5 -1.45 215.0 -0.61
60 265.3 -0.05 267.6 0.51 261.5 0.70 261.0 0.65
70 308.2 -0.40 310.6 0.05 303.5 0.22 301.8 -0.16
80 349.8 -1.05 351.6 -0.86 344.2 -0.50 302.0 -12.54
90 388.7 -2.23 388.8 -2.52 381.8 -1.85 378.9 -2.42
100 418.4 -5.26 424.9 -4.08 412.3 -4.59 408.3 -5.35

Conclusion: The results obtained with reagents stored for even 27 months under real-time conditions. From the
above results the stability claim of 24 months from the date of manufacture has been confirmed.

Design Verification and Product Data for GLUCOSE LIQUICOLOR 7/7

Rev. 006