lipase liquicolor · 2021. 2. 19. · humastar 600 human 16660 au 400 beckman coulter n1254600 au...

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


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


Day LOT 1 881012 LOT 2 881100


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





Sample 3 (high), LOT ERO172


Day LOT 1 881012 LOT 2 881100


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


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





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


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 % ✓


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



881100


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


1 CLSI. Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second

Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2012


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


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)


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


4 5.5585 7.9933 9.0721 4.6132 5.4822


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)


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


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


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



C.f.a.s CHEMA 10759350190 30558801

Sample HUMAN’s and commercial controls (DiaSys, RANDOX, CHEMA)


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.


Criteria

Passing and Bablok Acceptance criteria

Slope 0.95 – 1.05

Correlation coefficient r ≥ 0.95

Used Material


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)


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


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


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


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



Mean deviation for stressed and fresh reagent ≤ 10.0%

Used Material



C.f.a.s ROCHE 10759350190 30558801

Sample HUMAN’s and commercial controls (RANDOX, DiaSys, CHEMA)


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.





Criteria


From To Value Unit

0 60 16 U/l

60 332 27 %


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


Results

High pool

[%]



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.


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



Used Material



AUTOCAL HUMAN e 13060 0017

Sample HUMAN’s and commercial controls (RANDOX, DiaSys, CHEMA)


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.






Criteria


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


Results

High pool

[%]



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.


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


Max. Interference ± 10%

Used Material




Samples Samples spiked with interfering substance


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)


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


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)


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)


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


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.

lipase liquicolor · 2021. 2. 19. · humastar 600 human 16660 au 400 beckman coulter n1254600 au...

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