Upload
others
View
6
Download
0
Embed Size (px)
Citation preview
Design Verification LIPASE liquicolor Multipurpose Reagent
Form: 4.2-01.06-04 1/18 Rev. 004 | valid of 14.01.2021 .
Contents
1 Introduction .................................................................................................................................................................. 2
2 Imprecision .................................................................................................................................................................... 2
3 Linearity and Analytical sensitivity ............................................................................................................................. 4
3.1 Linearity ....................................................................................................................................................................................... 4
3.2 Analytical sensitivity ............................................................................................................................................................... 5
3.3 Measuring range ...................................................................................................................................................................... 8
4 Recovery of Control Sera .............................................................................................................................................. 8
5 Comparison of Methods ............................................................................................................................................... 8
6 Accelerated Stability testing...................................................................................................................................... 12
6.1 Recovery of Control sera ...................................................................................................................................................... 12
6.2 Linearity ..................................................................................................................................................................................... 12
7 Real Time Stability ...................................................................................................................................................... 13
8 Open Vial Stability ...................................................................................................................................................... 14
8.1 Recovery of control sera ...................................................................................................................................................... 14
8.2 Linearity ..................................................................................................................................................................................... 14
9 Interferences ............................................................................................................................................................... 16
10 Traceability .................................................................................................................................................................. 18
Design Verification and Product Data for Lipase 2/18 Rev. 004
1 Introduction
The performance characteristics of LIPASE liquicolor reagent has been tested and documented in order to verify the
clinical usefulness and compliance with the essential requirements of directive 98/79/EC.
Tested kit sizes
Reagent REF Content
LIPASE liquicolor
multipurpose reagent
12006 4 x 10 ml
12026 6 x 20 ml
LIPASE liquicolor system reagent 12006600 2 x 100 tests
Identical reagent formulations are used for the multipurpose and system reagents, therefore the results for the
system reagent are transferable to the multipurpose reagent. One of the multipurpose reagent kits was used for
the measurements on AU analysers, the measurements on the HumaStar 600 were carried out with system
reagent kit.
Used Devices
Analyzer Manufacturer REF
HumaStar 600 HUMAN 16660
AU 400 Beckman Coulter N1254600
AU 480 Beckman Coulter N3660400
2 Imprecision
The imprecision (repeatablility and within-lab / inter-assay) of LIPASE liquicolor reagent on HumaStar 600 was
calculated from 2 runs in duplicate on 20 consecutive days. For this study, 3 serum pools and 2 reagent LOTs were
employed. Calibration was performed before run. The means, standard deviations and coefficients of variation
were calculated.
Criteria
Imprecision Acceptance criteria
Repeatability CV 5.0%
Within-lab / inter-assay CV 8.0%
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12006600* 881012
881100
AUTOCAL HUMAN 13160 0017
Samples Low, medium and high pool sera LOT ERO171, 20200708-501-600,
ERO172
*manually filled bottles
Results
Sample 1 (low), LOT ERO171
Lipase liquicolor reagent
Day LOT 1 881012 LOT 2 881100
Mean Run 1 (U/l) Mean Run 2 (U/l) Mean Run 1 (U/l) Mean Run 2 (U/l)
1 23.7 25.3 23.9 26.6
2 23.7 25.9 24.1 25.2
3 24.1 25.2 24.6 26.3
4 25.5 25.4 23.9 24.0
5 23.6 23.7 26.1 26.2
6 26.2 26.4 26.0 25.1
7 26.6 25.6 26.5 24.7
8 26.3 29.5 25.6 27.3
9 27.0 27.5 26.9 26.7
10 26.2 26.8 26.0 26.9
11 24.9 24.2 26.6 28.6
Design Verification and Product Data for Lipase 3/18 Rev. 004
12 28.7 28.8 28.9 29.2
13 28.2 27.6 31.3 29.8
14 26.6 25.8 25.8 26.4
15 27.1 26.0 25.9 27.1
16 28.1 26.9 28.0 28.2
17 25.3 26.0 26.6 28.3
18 26.9 29.1 25.6 29.2
19 28.8 29.8 25.2 25.1
20 24.6 27.7 24.9 25.1
Mean 26.4 26.5
Repeatability - SD 0.98 0.65
Repeatability - %CV 3.7 2.5
Within-lab - SD 1.83 1.80
Within-lab - %CV 6.9 6.8
Sample 2 (medium), LOT 20200708-501-600
Lipase liquicolor reagent
Day LOT 1 881012 LOT 2 881100
Mean Run 1 (U/l) Mean Run 2 (U/l) Mean Run 1 (U/l) Mean Run 2 (U/l)
1 47.3 49.7 48.9 52.3
2 48.5 48.5 47.8 51.1
3 47.4 50.1 52.0 54.1
4 49.8 48.9 51.2 50.0
5 47.8 49.8 51.4 51.3
6 52.1 55.1 52.4 52.5
7 51.6 49.6 52.2 53.0
8 52.3 51.1 52.4 54.9
9 52.0 52.0 54.1 53.5
10 53.2 52.7 52.4 54.8
11 48.7 48.8 55.9 57.2
12 53.8 56.3 56.5 60.1
13 53.6 54.4 58.0 58.3
14 52.6 49.8 53.7 54.4
15 50.8 50.5 52.3 56.1
16 52.7 52.2 53.7 58.3
17 49.8 53.7 56.0 56.7
18 49.4 52.5 51.6 55.0
19 47.9 50.5 50.6 52.0
20 49.7 53.3 50.8 54.6
Mean 51.0 53.6
Repeatability - SD 1.14 1.70
Repeatability - %CV 2.2 3.2
Within-lab - SD 2.38 3.00
Within-lab - %CV 4.7 5.6
Sample 3 (high), LOT ERO172
Lipase liquicolor reagent
Day LOT 1 881012 LOT 2 881100
Mean Run 1 (U/l) Mean Run 2 (U/l) Mean Run 1 (U/l) Mean Run 2 (U/l)
1 143.3 159.6 157.0 169.5
2 149.8 149.9 161.7 158.0
3 143.6 145.1 175.3 157.6
4 153.0 146.5 162.9 156.2
5 146.8 146.9 158.0 156.5
6 147.1 146.7 149.7 150.7
7 146.8 147.8 155.3 154.6
8 152.7 152.9 161.1 157.5
Design Verification and Product Data for Lipase 4/18 Rev. 004
9 150.2 151.8 161.2 159.5
10 147.9 146.6 154.3 157.0
11 140.8 145.6 148.9 157.2
12 149.5 148.3 159.2 160.7
13 153.8 148.7 163.1 167.4
14 150.4 146.5 162.6 154.3
15 146.5 151.2 161.8 158.7
16 140.2 148.6 160.8 156.5
17 146.8 150.8 160.4 154.9
18 147.8 150.1 164.3 158.2
19 144.3 151.9 157.8 160.5
20 139.4 141.9 155.4 151.4
Mean 148 159
Repeatability - SD 2.78 2.67
Repeatability - %CV 1.9 1.7
Within-lab - SD 4.45 5.45
Within-lab - %CV 3.0 3.4
Conclusion
The LIPASE liquicolor reagent on HumaStar 600 complies with the acceptance criteria for repeatability and within-
lab/inter-assay imprecision for low, medium and high concentrated serum pools: <5.0% and <8.0%, respectively.
3 Linearity and Analytical sensitivity
3.1 Linearity
The linearity of LIPASE liquicolor reagent was controlled by employing a high concentrated linearity pool
successively diluted in different steps with physiological saline. The measurements were carried out on a HumaStar
600. The analysed concentrations were compared with the theoretical concentrations obtained from a linear
regression.
The linearity of the test was evaluated over the difference between best fitting nonlinear polynomial and linear
polynomial. In this case difference from the 2nd to 1st order was used. In accordance with the European Society for
External Quality Assessment, the specification is based on a deviation of up to 16 U/l to 60 U/l Lipase and up to 27%
from a concentration from 60 U/l Lipase.
Criteria
Range limit Acceptance criteria max. Deviation from linearity
From To Value Unit
0 60 16 U/l
60 330 27 %
Reagent Manufacturer REF LOT LIPASE liquicolor reagent HUMAN 12006600* 881012 AUTOCAL HUMAN 13160 0017
Sample High linearity pool, LOT ERO180, physiological saline, LOT MPE005
*manually filled bottles
Results
High pool
[%]
Mean Predicted Polynominal Difference (2nd – 1st) Specification
U/l 1st order 2nd order U/l % ≤ ✓
0 3.8 2.3 3.4 1.1 29.2 16 U/l ✓
0.5 4.9 3.9 4.9 1 20.4 16 U/l ✓
1 5 5.4 6.4 1 20.1 16 U/l ✓
2 6.8 8.5 9.3 0.8 11.8 16 U/l ✓
5 18.5 17.7 18.2 0.5 2.7 16 U/l ✓
10 34.7 33.1 33 -0.1 -0.3 16 U/l ✓
20 66.4 63.8 62.9 -0.9 -1.4 27 % ✓
30 90.8 94.5 93.1 -1.4 -1.5 27 % ✓
Design Verification and Product Data for Lipase 5/18 Rev. 004
40 125.1 125.3 123.5 -1.8 -1.4 27 % ✓
50 160 156 154.3 -1.7 -1.1 27 % ✓
60 180 186.8 185.3 -1.5 -0.8 27 % ✓
70 213.4 217.5 216.6 -0.9 -0.4 27 % ✓
80 242.2 248.2 248.2 0 0.0 27 % ✓
90 290.5 279 280.1 1.1 0.4 27 % ✓
100 309.6 309.7 312.3 2.6 0.8 27 % ✓
Graphic
Conclusion
The LIPASE liquicolor reagent on HumaStar 600 is linear from 5 to 300 U/l.
3.2 Analytical sensitivity
A method according to CLSI EP 17 2nd edition1 was used to verify the limits of blank, detection and quantification.
The tests on the HumaStar 600 were carried out for 2 reagent LOTs each.
The limit of blank (LoB) was evaluated on 3 days using a 4-fold determination of 5 blank samples (n = 60).
The limit of detection (LoD) was evaluated on 3 days using a 4-fold determination of 5 low concentrated serum
samples (n=60).
The limit of quantification (LoQ) was evaluated on 3 days using a 3-fold determination of 5 low concentrated serum
samples (n = 45). The determination of the LoQ based on the total error (%TE). The reference value was determined
with a commercially available lipase calibrator on HumaStar 600. The total error claim (30%) refers to Ricos Goals
2014.
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12006600* 881012
881100
AUTOCAL HUMAN 13160 0017
C.f.a.s (reference) ROCHE 10759350190 30558801
Samples for LoB 5 blank samples (physiological saline), LOTs MPE005C, MPR005B, MPE005A,
MPE005, 20200401-001
Samples for LoD 5 low samples, LOTs ERO173, ERO174, ERO175, ERO176, ERO177
Samples for LoQ 5 low samples, LOTs BOG440, BOG441, BOG442, BOG443, BOG444
*manually filled bottles
1 CLSI. Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second
Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2012
Design Verification and Product Data for Lipase 6/18 Rev. 004
Results
Limit of Blank (LoB)
Day Replicate
Blank 1
LOT MPE005C
Blank 2
LOT MPR005B
Blank 3
LOT MPE005A
Blank 4
LOT MPE005
Blank 5
LOT 20200401-
001
(U/l) (U/l) (U/l) (U/l) (U/l)
Results for LIPASE liquicolor reagent, LOT 881012
1
1 -0.7890 4.0149 -0.2440 1.2156 -0.7802
2 -0.8293 3.2537 -0.3058 0.7418 -0.7700
3 -1.0844 -0.8318 1.9015 1.3299 -1.0604
4 -0.7925 -0.8334 2.2210 1.3755 -0.7758
2
1 2.2225 -0.5704 0.0326 -0.6548 -0.8769
2 -0.2526 0.2325 0.9236 -0.6695 -0.6635
3 -0.4032 0.1081 0.4474 -0.5642 -0.8209
4 -0.3263 0.5303 -0.7477 -0.3740 -1.0561
3
1 -0.1296 0.2373 -0.0606 -0.2136 0.4901
2 -0.0790 -0.2478 -0.3038 -0.2975 0.3190
3 -0.1539 -0.3000 -0.0182 -0.2799 -0.0193
4 0.1193 -0.2680 -0.2008 -0.0730 -0.2968
Results for LIPASE liquicolor reagent, LOT 881100
1
1 -2.3162 -3.3330 -1.1950 -3.0698 -3.3956
2 -0.0873 1.9236 -2.6246 -3.0714 -3.2105
3 -0.3350 4.7899 -2.9718 -2.9749 -2.0161
4 -1.0320 3.7164 -3.0286 -3.0463 -1.9672
2
1 0.3676 0.5976 -1.0072 -0.6711 -0.4923
2 0.1385 0.9670 -1.0070 -0.7927 -0.3992
3 1.8157 -0.6649 -1.0531 -0.8948 0.7032
4 1.8187 -0.6421 -0.9727 -0.1715 -0.0021
3
1 -0.3373 -0.6554 -0.9877 -0.5844 -0.6093
2 -0.5531 -0.7384 -0.9807 -0.4516 -0.7535
3 -0.6328 -0.7330 -0.7614 -0.5946 0.1397
4 -0.5479 -0.6870 -0.7455 -0.7020 1.1649
Calculated LoB
LOT 1 1.7 U/l
LOT 2 1.9 U/l
Reported LoB 1.9 U/l
Limit of Detection (LoD)
Day Replicate LoD sample 1
LOT ERO173
LoD sample 2
LOT ERO174
LoD sample 3
LOT ERO175
LoD sample 4
LOT ERO176
LoD sample 5
LOT ERO177
(U/l) (U/l) (U/l) (U/l) (U/l)
Results for LIPASE liquicolor reagent, LOT 881012
1
1 10.4157 8.3338 10.0889 4.7058 8.6178
2 10.0632 7.9969 9.6683 6.9577 4.7077
3 5.5176 9.1327 9.5049 5.7566 4.6554
4 6.1568 8.8810 9.3798 3.8565 6.4862
2
1 5.3467 10.7140 11.5912 4.5981 4.9040
2 6.1981 7.3095 10.2219 4.0572 5.1831
3 5.7413 6.8792 9.0943 4.6578 5.2034
4 6.2348 6.5921 8.7915 4.1850 5.1915
3
1 5.5662 10.7419 12.1819 4.5728 5.2552
2 6.8303 9.9302 9.1941 4.4244 5.5441
3 6.8572 11.1044 9.2972 4.6948 5.3857
Design Verification and Product Data for Lipase 7/18 Rev. 004
4 5.5585 7.9933 9.0721 4.6132 5.4822
Results for LIPASE liquicolor reagent, LOT 881100
1
1 2.7830 11.2490 6.3682 1.2831 4.6451
2 4.6799 4.0872 6.3260 1.2635 3.4272
3 4.3707 4.2377 7.8614 5.0092 2.4392
4 7.6836 4.1274 7.7417 5.6949 2.1773
2
1 3.8594 6.1578 8.5490 4.5449 4.0156
2 3.7281 5.9772 8.9309 4.9488 3.8570
3 3.3577 6.2148 9.0323 4.7340 3.9838
4 4.2796 7.1100 10.4216 3.6375 4.0845
3
1 4.7367 7.6809 8.9246 4.1367 4.6889
2 5.0576 7.0279 9.4660 3.9883 4.6441
3 4.9367 8.1218 9.2653 3.9954 4.3975
4 5.6028 8.2302 8.2733 4.0455 4.5626
Calculated LoD
LOT 1 4.0 U/l
LOT 2 4.2 U/l
Reported LoD 4.2 U/l
Limit of Quantification (LoQ)
Day Replicate LoQ sample 1 LoQ sample 2 LoQ sample 3 LoQ sample 4 LoQ sample 5
(U/l) (U/l) (U/l) (U/l) (U/l)
Results for LIPASE liquicolor reagent, LOT 881012
1
1 9.4386 6.8860 7.4867 17.4297 11.2892
2 10.1888 7.0865 7.8118 16.5567 11.3470
3 10.0975 6.6182 9.2401 10.7008 12.4694
2
1 9.3032 7.2482 6.0308 9.9945 10.6427
2 8.9853 6.9398 5.7674 9.9729 11.4595
3 8.5078 7.2762 6.1464 9.8416 10.4561
3
1 9.2982 7.1896 5.9330 12.4924 10.8797
2 9.1197 7.0307 6.2309 11.4243 10.8915
3 9.1539 6.8500 6.0091 11.6983 11.2247
Results for LIPASE liquicolor reagent, LOT 881100
1
1 6.1225 3.8597 4.2049 7.7718 10.4270
2 6.0937 3.6401 4.1216 10.4779 7.0343
3 6.3315 3.8439 3.7725 9.9957 7.0386
2
1 7.9148 6.7150 4.8822 8.6536 9.5725
2 8.8319 6.8122 4.7503 8.6486 9.6007
3 8.8434 6.9920 4.7539 8.7160 10.7215
3
1 8.5763 6.4616 5.5318 9.6658 10.8530
2 8.3813 6.7340 5.1230 9.6625 10.2336
3 8.5938 6.7700 5.8275 10.2854 10.5093
Reagent LOQ sample no. Reference value (U/l) Mean (U/l) TE%
LOT 1 2 6.7 7.0 11
LOT 2 4 10.7 9.3 30
Reported LoQ 9.3
Design Verification and Product Data for Lipase 8/18 Rev. 004
Conclusion
The limit of blank (LoB), the limit of detection (LoD) and the limit of quantificatin (LoQ) were determined for 2 LOTs
of LIPASE liquicolor reagent on HumaStar 600 as follows:
LoB (U/l) LoD (U/l) LoQ (U/l)
HumaStar 600 1.9 4.2 9.3
3.3 Measuring range
The measuring range for LIPASE liquicolor reagent on HumaStar 600 was determined taking into account the
linearity range and the LoQ: 10 – 300 U/l.
4 Recovery of Control Sera
A number of commercially available control sera were employed on HumaStar 600. The control sera were
reconstituted/prepared according to the manufacturer’s instructions. The controls were measured in duplicate. The
recovery within the respective allowable range was checked and the recovery of the respective target value was
calculated.
Criteria
Check Acceptance criteria
Recovery within range
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12006600* 881012
C.f.a.s CHEMA 10759350190 30558801
Sample HUMAN’s and commercial controls (DiaSys, RANDOX, CHEMA)
*manually filled bottles
Results Control recovery Name LOT Target
U/L Range
U/L Result
U/L Within range
YES/NO Recovery
%
SERODOS 0004 42.1 33.7 – 50.5 40.4 39.0
YES YES
96 93
SERODOSplus 0005 63.0 50.4 – 75.6 57.8 57.7
YES YES
92 92
HUM ASY 2 1308UN 33.0 27.0 – 39.0 28.8 32.1
YES YES
87 97
HUM ASY 3 1032UE 61.0 49.0 – 73.0 71.1 72.1
YES YES
117 118
TruLab N 27547 51.0 40.8 – 61.2 58.1 53.5
YES YES
106 97
TruLab P 27548 100 80.1 - 120 104 103
YES YES
98 97
Quantinorm 730225 53.5 44.5 – 62.5 48.2 52.9
YES YES
90 99
Quantipath 621177 112 91.0 - 133 104 100
YES YES
93 89
Conclusion
Recovery for SERODOS controls is very good; all controls are measured within the allowable ranges. Therefore, the
control recovery verification for the LIPASE liquicolor reagent on HumaStar 600 is accepted.
5 Comparison of Methods
LIPASE liquicolor reagent (test) was compared with Lipase DC FS unversal reagent, DiaSys (reference). Patient
samples (serum and heparin plasma; n = 127) were included to the comparison. The measurements were carried
out on an AU 480.
The results were evaluated by a non-parametric regression analysis according to Passing and Bablok.
Design Verification and Product Data for Lipase 9/18 Rev. 004
Criteria
Passing and Bablok Acceptance criteria
Slope 0.95 – 1.05
Correlation coefficient r ≥ 0.95
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent (test) HUMAN 12006* 881012
AUTOCAL (test) HUMAN 13160 0017
Lipase DC FS (reference) DiaSys 143219910021 60139232
TruCal U (reference) DiaSys 591009910064 26474
Samples Patient samples (serum and heparin plasma)
*manually filled bottles
Results
Sample-no. UNIT Mean Reference
Lipase DC FS, DiaSys
Mean Test
LIPASE liquicolor, HUMAN Difference
1 U/L 21.9 26.8 4.8
2 U/L 24.7 31.0 6.3
3 U/L 69.6 78.1 8.5
4 U/L 36.6 44.1 7.5
5 U/L 31.7 39.2 7.6
6 U/L 24.8 28.8 4.0
7 U/L 30.1 36.4 6.3
8 U/L 40.3 46.0 5.8 9 U/L 39.9 44.0 4.1
10 U/L 37.2 42.8 5.6 11 U/L 28.2 36.0 7.8 12 U/L 33.9 40.2 6.3 13 U/L 31.8 39.1 7.3 14 U/L 19.7 26.6 6.9 15 U/L 27.5 32.5 5.0 16 U/L 35.1 39.5 4.4 17 U/L 20.1 27.7 7.6 18 U/L 36.7 43.3 6.5 19 U/L 32.7 37.9 5.2 20 U/L 32.9 41.1 8.2 21 U/L 27.8 29.8 2.0 22 U/L 27.8 31.0 3.2 23 U/L 35.3 35.2 -0.1 24 U/L 21.3 25.5 4.3 25 U/L 82.9 87.2 4.2 26 U/L 30.6 38.4 7.8 27 U/L 32.4 35.3 2.9 28 U/L 30.8 38.1 7.3 29 U/L 69.5 75.2 5.7 30 U/L 62.5 67.9 5.4 31 U/L 36.3 33.5 -2.7 32 U/L 36.4 41.4 4.9 33 U/L 54.0 54.4 0.4 34 U/L 54.6 55.9 1.3 35 U/L 17.1 22.3 5.2 36 U/L 31.8 36.0 4.3 37 U/L 24.7 25.9 1.2 38 U/L 51.6 53.9 2.4 39 U/L 40.4 43.1 2.8 40 U/L 36.6 35.0 -1.6 41 U/L 37.7 41.7 4.1 42 U/L 28.8 34.5 5.7 43 U/L 24.5 28.9 4.4
Design Verification and Product Data for Lipase 10/18 Rev. 004
44 U/L 48.8 52.9 4.1 45 U/L 39.5 47.1 7.7 46 U/L 29.6 31.1 1.5 47 U/L 21.1 25.3 4.2 48 U/L 11.0 15.4 4.5 49 U/L 66.2 65.1 -1.1 50 U/L 23.1 26.9 3.9 51 U/L 22.4 28.1 5.7 52 U/L 136 115 -20.8 53 U/L 24.9 27.6 2.7 54 U/L 46.1 47.3 1.1 55 U/L 48.7 40.1 -8.6 56 U/L 29.2 37.4 8.2 57 U/L 18.3 22.1 3.8 58 U/L 19.7 26.2 6.5 59 U/L 24.7 28.6 3.9 60 U/L 36.7 37.5 0.8 61 U/L 21.6 28.7 7.1 62 U/L 43.5 41.4 -2.1 63 U/L 41.7 50.6 8.9 64 U/L 37.2 42.7 5.5 65 U/L 36.8 36.1 -0.7 66 U/L 21.9 26.4 4.5 67 U/L 44.5 42.3 -2.2 68 U/L 42.3 46.6 4.3 69 U/L 51.0 58.3 7.4 70 U/L 36.1 36.1 0.1 71 U/L 47.3 52.8 5.5 72 U/L 28.0 33.7 5.7 73 U/L 37.5 37.3 -0.2 74 U/L 33.8 34.1 0.3 75 U/L 23.1 30.5 7.4 76 U/L 45.2 51.5 6.2 77 U/L 47.1 48.7 1.6 78 U/L 39.4 39.6 0.3 79 U/L 74.5 75.4 0.9 80 U/L 36.7 42.0 5.3 81 U/L 33.4 37.7 4.4 82 U/L 27.0 32.9 5.8 83 U/L 39.7 45.2 5.5 84 U/L 23.4 29.7 6.3 85 U/L 267 283 15.3 86 U/L 123 116 -6.7 87 U/L 281 289 7.8 88 U/L 264 223 -40.4 89 U/L 332 329 -3.2 90 U/L 213 221 8.2 91 U/L 254 172 -81.7 92 U/L 253 251 -2.0 93 U/L 199 187 -12.6 94 U/L 273 219 -54.2 95 U/L 287 287 -0.2 96 U/L 270 221 -48.5 97 U/L 308 304 -4.0 98 U/L 45.4 53.9 8.5 99 U/L 15.6 28.3 12.6
100 U/L 130 143 13.5 101 U/L 50.7 55.6 5.0 102 U/L 21.1 30.4 9.3 103 U/L 34.3 44.1 9.9 104 U/L 30.1 44.3 14.2
Design Verification and Product Data for Lipase 11/18 Rev. 004
105 U/L 26.5 43.8 17.3 106 U/L 18.5 33.0 14.5 107 U/L 19.0 33.6 14.5 108 U/L 36.9 50.3 13.4 109 U/L 111 124 12.3 110 U/L 189 199 10.4 111 U/L 115 116 1.5 112 U/L 112 116 3.8 113 U/L 153 161 7.9 114 U/L 164 176 12.4 115 U/L 123 139 15.7 116 U/L 200 215 15.0 117 U/L 121 131 9.3 118 U/L 95.9 105 8.8 119 U/L 245 224 -21.5 120 U/L 156 153 -2.9 121 U/L 250 177 -73.8 122 U/L 222 218 -4.8 123 U/L 222 223 1.2 124 U/L 286 261 -24.6 125 U/L 265 211 -53.8 126 U/L 212 193 -18.8 127 U/L 79.4 82.6 3.2
N 127 127
Mean U/L 80.2 81.2
Min U/L 11.0 15.4
Max U/L 332 329
Graphic
Y = 0.9648 X – 6.4976 (U/l) r = 0.9872 Y = test X = reference
Conclusion
LIPASE liquicolor reagent shows a good agreement with Lipase DC FS, DiaSys, on AU 480
(r = 0.9872), and no significant deviation could be observed with any specific sample:
Y = 0.9648 X – 6.4976 (U/l)
Y = test. X = reference
Design Verification and Product Data for Lipase 12/18 Rev. 004
6 Accelerated Stability testing
6.1 Recovery of Control sera
The recovery of control sera was tested according to the procedure already described in section 4 on AU 400. The
reagent was stressed in multipurpose reagent bottles for 10 days at 37°C. The mean values (n=3) obtained with
fresh reagent (=reference) and stressed reagent were calculated and compared.
Criteria
Check Acceptance criteria
Recovery within range
Mean deviation for stressed and fresh reagent ≤ 10.0%
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12026* 881012
C.f.a.s ROCHE 10759350190 30558801
Sample HUMAN’s and commercial controls (RANDOX, DiaSys, CHEMA)
*manually filled bottles
Results
LIPASE liquicolor, LOT 881012 Fresh 10 days 37°C
Name LOT Target
U/l
Range
U/l
Result
U/l
Within
range
YES/NO
Dev. to
Target
%
Result
U/l
Within range
YES/NO
Dev. to
Fresh
%
SERODOS 0004 42.1 33.7 - 50.5 45.6 YES 8.4 46.6 YES 2.2
SERODOSplus 0005 63.0 50.4 – 75.6 68.2 YES 8.3 73.8 YES 8.2
HUM ASY 2 1308UN 33.0 27.0 – 39.0 38.8 YES 17.6 37.7 YES -2.8
HUM ASY 3 1032UE 61.0 49.0 – 73.0 63.3 YES 3.7 66.6 YES 5.2
TruLab N 24382 54.9 43.9 – 65.9 63.7 YES 16.1 61.9 YES -2.8
TruLab P 24383 107 85.6-- 128 108 YES 1.1 104 YES -3.9
Quantinorm 730225 53.5 44.5 – 62.5 58.3 YES 8.9 57.4 YES -1.5
Quantipath 621177 112 91.0 - 133 116 YES 3.2 111 YES -3.7
Dev.% 8.4 0.1
Recovery of controls for LIPASE liquicolor reagent stored at 37°C for 10 days is within the acceptance criteria.
6.2 Linearity
The linearity of LIPASE liquicolorreagent was investigated according to the procedure already described in section
3.1 using a high concentrated patient pool successively diluted in different steps with a low concentrated patient
pool. The measurements were carried out on an AU 400. The analysed concentrations were compared with the
theoretical concentrations obtained from a linear regression.
The linearity of the test was evaluated over the difference between best fitting nonlinear polynomial and linear
polynomial. In this case difference from the 2nd to 1st order was used. In accordance with the European Society for
External Quality Assessment, the specification is based on a deviation of up to 16 U/l to 60 U/l Lipase and up to 27%
from a concentration from 60 U/l Lipase.
Criteria
Range limit Acceptance criteria max. Deviation from linearity
From To Value Unit
0 60 16 U/l
60 332 27 %
Design Verification and Product Data for Lipase 13/18 Rev. 004
Used Material
Reagent Manufacturer REF LOT LIPASE liquicolor reagent HUMAN 12026* 881012
C.f.a.s ROCHE 10759350190 30558801
Sample High pool sample, LOT 190813328-400, low pool sample, LOT
20200312BOG001
*manually filled bottles
Results
High pool
[%]
Mean Predicted Polynominal Difference (2nd – 1st) Specification
U/l 1st order 2nd order U/l % ≤ ✓
0 38.45 42.77 38.02 -4.75 -12.4 16 U/l ✓
0.5 40 44.52 40.04 -4.48 -11.2 16 U/l ✓
1 41.99 46.26 42.05 -4.21 -10.0 16 U/l ✓
2 45.84 49.75 46.08 -3.67 -8.0 16 U/l ✓
5 57.73 60.21 58.07 -2.14 -3.7 27 % ✓
10 78.97 77.65 77.81 0.16 0.2 27 % ✓
20 114.7 112.52 116.37 3.85 3.4 27 % ✓
30 154.31 147.39 153.69 6.3 4.1 27 % ✓
40 189.06 182.27 189.78 7.51 4.0 27 % ✓
50 225.73 217.14 224.63 7.49 3.3 27 % ✓
60 258.12 252.01 258.25 6.24 2.4 27 % ✓
70 291.57 286.88 290.63 3.75 1.3 27 % ✓
80 321.58 321.76 321.78 0.02 0.0 27 % ✓
90 349.7 356.63 351.7 -4.93 -1.4 > high limit
100 381.56 391.5 380.38 -11.12 -2.9 >high limit
Graphic
After 10 days at 37°C the LIPASE liquicolor reagent is linear up to 300 U/l.
Conclusion
Accelerated stress data for LIPASE liquicolor reagent, REF 12006 and 12026, are indicative for a real-time stability of
18 months at a storage temperature of 2...8°C.
7 Real Time Stability
Real time stability studies of LIPASE liquicolor reagent are ongoing.
Design Verification and Product Data for Lipase 14/18 Rev. 004
8 Open Vial Stability
8.1 Recovery of control sera
For the opened reagent stability study, LIPASE liquicolor reagent was tested for 95 days. Recovery of control sera
was tested according to the procedure already described in section 4.
Measurements were performed at several intervals on an AU 480. After each use, the reagent was properly sealed
and stored again at 2...8°C.
Criteria
Check Acceptance criteria
Recovery within range
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12026* 881012
AUTOCAL HUMAN e 13060 0017
Sample HUMAN’s and commercial controls (RANDOX, DiaSys, CHEMA)
*manually filled bottles
Results
Controls Freshly opened After 95 days
Name LOT Target
U/l
Range
U/l
Result
U/l
Within range
YES/NO
Result
U/l
Within range
YES/NO
SERODOS 0005 49.2 39.4 – 59.0
49.9 47.6 49.6
YES 48.9 48.7 51.4
YES
SERODOSplus 0005 63.0 50.4 – 75.6
67.0 66.9 67.3
YES 64.1 63.2 68.2
YES
TruLab N 57547 51.0 40.8 – 61.2
58.9 53.3 54.8
YES 55.2 54.4 54.5
YES
TruLab P 27548 100 80.1 – 120
104 101 109
YES 100 98.5 102
YES
HUM ASY 2 1308UN 33.0 27.0 – 39.0
34.2 31.6 34.6
YES 32.0 33.6 35.8
YES
HUM ASY 3 1032UE 61.0 49.0 – 73.0
59.5 61.5 59.0
YES 57.7 56.3 58.4
YES
Quantinorm 881068 41.6 32.6 – 50.6
44.6 44.8 46.1
YES 46.0 45.9 47.2
YES
Quantipath 621177 112 91.0 - 133
118 116 111
YES 112 114 111
YES
95 days after opening the control recovery for the LIPASE liquicolor reagent was within the respective allowable
range.
8.2 Linearity
The linearity of LIPASE liquicolor reagent was investigated according to the procedure already described in section
3.1 using a high concentrated patient pool successively diluted in different steps with with physiological saline. The
measurements were carried out on an AU 400. The analysed concentrations were compared with the theoretical
concentrations obtained from a linear regression.
The linearity of the test was evaluated over the difference between best fitting nonlinear polynomial and linear
polynomial. In this case difference from the 2nd to 1st order was used. In accordance with the European Society for
External Quality Assessment, the specification is based on a deviation of up to 16 U/l to 60 U/l Lipase and up to 27%
from a concentration from 60 U/l Lipase.
Design Verification and Product Data for Lipase 15/18 Rev. 004
Criteria
Range limit Acceptance criteria max. Deviation from linearity
From To Value Unit
0 60 16 U/l
60 350 27 %
Used Material
Reagent Manufacturer REF LOT LIPASE liquicolor reagent HUMAN 12026* 881012 AUTOCAL HUMAN 13160 0017
Sample High linearity pool, LOT MZM014, physiological saline, LOT BER234
*manually filled bottles
Results
High pool
[%]
Mean Predicted Polynominal Difference (2nd – 1st) Specification
U/l 1st order 2nd order U/l % ≤ ✓
0 0.2 0.1 -5.3 -5.4 -2347.8 16 U/l ✓
0.5 -0.2 2.2 -3 -5.2 2311.1 16 U/l ✓
1 3.3 4.2 -0.6 -4.8 -144.6 16 U/l ✓
2 2.2 8.3 4.1 -4.2 -192.2 16 U/l ✓
5 7.6 20.7 18.2 -2.5 -33.1 16 U/l ✓
10 38.3 41.3 41.5 0.2 0.5 16 U/l ✓
20 88.9 82.4 86.8 4.4 4.9 27 % ✓
30 130.9 123.6 130.8 7.2 5.5 27 % ✓
40 173.1 164.7 173.4 8.7 5.0 27 % ✓
50 215.7 205.9 214.5 8.6 4.0 27 % ✓
60 258.3 247.1 254.2 7.1 2.7 27 % ✓
70 290.7 288.2 292.5 4.3 1.5 27 % ✓
80 329.2 329.4 329.4 0 0.0 27 % ✓
90 360.7 370.5 364.9 -5.6 -1.6 > high limit
100 401.6 411.7 398.9 -12.8 -3.2 > high limit
Graphic
95 days after opening the LIPASE liquicolor reagent is linear from 1 to 300 U/l.
Conclusion
After first opening and subsequent storage at 2 – 8 °C, the LIPASE liquicolor reagents, REF 12006, 12026, are stable
for at least 90 days if properly sealed after each use.
Design Verification and Product Data for Lipase 16/18 Rev. 004
9 Interferences
A method according to CLSI EP 7 3rd edition2 and EP 37 1st edition (supplemental tables)3 was used to verify the
interference of LIPASE liquicolor reagent. The interference has been studied by adding known amounts of the
potentially interfering substance to samples with known low and high lipase concentrations (analyte). The
measurements were carried out on a HumaStar 600.
Criteria
Check Acceptance criteria
Max. Interference ± 10%
Used Material
Reagent Manufacturer REF LOT
LIPASE liquicolor reagent HUMAN 12006600* 881012
AUTOCAL HUMAN 13160 0017
Samples Samples spiked with interfering substance
*manually filled bottles
Results for Bilirubin interference
Low Sample Added Bilirubin concentration (mg/dl)
Replicate Unit 0 10 20 30 40
1 U/l 48.9 47.9 47.6 47.7 47.9
2 U/l 48.7 48.5 47.3 47.6 47.9
3 U/l 49.5 47.3 48.2 47.9 47.6
4 U/l 49.1 48.2 47.9 47.1 48.8
5 U/l 47.4 48.2 48.3 46.9 48.5
Mean U/l 48.7 48.0 47.8 47.4 48.1
Tested effect % -1.4 -1.8 -2.6 -1.8
High sample Added Bilirubin concentration (mg/dl)
Replicate Unit 0 10 20 30 40
1 U/l 116 117 117 119 113
2 U/l 114 115 114 118 112
3 U/l 116 116 115 117 111
4 U/l 117 116 118 115 113
5 U/l 117 116 119 114 113
Mean U/l 116 116 117 117 112
Tested effect % 0.009 0.6 0.6 -3.0
Results for Hemoglobin interference
Low Sample Added Hemoglobin concentration (mg/dl)
Replicate Unit 0 250 500 750 1000
1 U/l 47.3 55.4 59.2 58.6 59.9
2 U/l 47.1 56.2 60.3 57.0 60.1
3 U/l 46.4 55.2 57.5 57.1 61.5
4 U/l 46.1 55.7 56.5 56.8 62.2
5 U/l 45.3 53.0 56.6 57.9 61.3
Mean U/l 46.4 55.1 58.0 57.5 61.0
Tested effect % 18.7 25.0 23.8 31.4
High sample Added Hemoglobin concentration (mg/dl)
Replicate Unit 0 250 500 750 1000
1 U/l 115 141 145 148 151
2 CLSI. Interference Testing in Clinical Chemistry. 3rd edition. CLSI Guideline EP 7. Wayne, PA: Clinical and Laboratory
Standards Institute; 2018 3 CLSI. Supplemental Tables for Interference Testing in Clinical Chemistry. 1st edition. CLSI Supplement EP 37, Wayne, PA:
Clinical and Laboratory Standards Institute; 2018
Design Verification and Product Data for Lipase 17/18 Rev. 004
2 U/l 122 140 147 147 150
3 U/l 123 142 144 145 151
4 U/l 125 139 144 147 152
5 U/l 124 140 143 146 154
Mean U/l 122 141 145 147 152
Tested effect % 15.3 18.7 20.4 24.4
Results for Intralipid interference
Low sample Added Intralipid concentration (mg/dl)
Replicate Unit 0 250 500 750 1000
1 U/l 52.8 40.7 35.9 32.0 30.5
2 U/l 54.8 39.8 33.8 32.8 29.1
3 U/l 54.5 39.9 34.4 32.5 29.9
4 U/l 55.4 39.8 34.2 31.1 29.8
5 U/l 53.3 38.3 33.5 32.5 30.0
Mean U/l 54.2 39.7 34.4 32.2 29.8
Tested effect % -26.7 -36.5 -40.6 -44.9
High sample Added Intralipid concentration (mg/dl)
Replicate Unit 0 250 500 750 1000
1 U/l 123 84.0 61.3 47.7 40.0
2 U/l 125 84.8 61.6 49.2 38.7
3 U/l 124 83.8 62.1 48.3 37.9
4 U/l 125 82.9 61.4 48.3 37.9
5 U/l 123 84.6 61.7 47.8 37.8
Mean U/l 124 84.0 61.6 48.2 38.5
Tested effect % -32.3 -50.3 -61.1 -69.0
Results for Ascorbic acid interference
Low sample Added Ascorbic acid concentration (mg/dl)
Replicate Unit 0 5 10 15 20
1 U/l 56.6 55.0 57.8 57.5 56.3
2 U/l 54.0 54.6 56.5 57.9 52.8
3 U/l 55.3 57.6 57.2 57.8 52.7
4 U/l 55.1 57.2 56.8 57.8 52.5
5 U/l 53.3 56.3 57.8 56.3 52.2
Mean U/l 54.9 56.1 57.2 57.4 53.3
Tested effect % 2.3 4.3 4.7 -2.9
High sample Added Ascorbic acid concentration (mg/dl)
Replicate Unit 0 5 10 15 20
1 U/l 125 121 125 145 135
2 U/l 123 121 145 142 134
3 U/l 122 117 145 140 135
4 U/l 122 119 143 139 137
5 U/l 121 123 143 139 138
Mean U/l 123 120 140 141 136
Tested effect % -1.9 14.2 14.9 10.6
Design Verification and Product Data for Lipase 18/18 Rev. 004
Conclusion
Up to the following concentrations, the interference of LIPASE liquicolor reagent was found to be ≤ 10% on
HumaStar 600:
Interfering substance Low Lipase concentration
(46.1 U/l) High Lipase concentration
(153 U/l)
Ascorbic acid up to 20 mg/dl up to 8.7 mg/dl
Bilirubin up to 40 mg/dl up to 40 mg/dl
Hemolysis (Hemoglobin) < 250 mg/dl
high interference < 250 mg/dl
high interference
Lipemia (Intralipid) < 250 mg/dl
high interference < 250 mg/dl
high interference
10 Traceability
LIPASE liquicolor reagent is calibrated with AUTOCAL REF 13160,which is calibrated against a commercial available
lipase calibrator.