LIPASE liquicolor. Design Verification. Multipurpose Reagent

Transcription

1 Design Verification LIPASE liquicolor Multipurpose Reagent CONTENTS 1 Introduction Imprecision Linearity and Detection Limit Linearity Detection Limit Recovery of Control Sera Comparison of Methods Stability, Accelerated Stress Linearity Recovery of Control Sera Real Time Stability Recovery of Control Sera Open Vial Stability Linearity Recovery of Control Sera Interferences Traceability... 9 Form: /9 valid of

2 1 Introduction The performance characteristics of LIPASE liquicolor have 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 LIPASE liquicolor ([BUF] lot and [SUB] , exp date ) was calculated from six determinations on five consecutive days. Control sera with low, medium and high lipase levels were employed as sample material. The tests were performed on AU 400 system. The mean, standard deviation and coefficient of variation were calculated. Analyte concentration Intra-assay Inter-assay (U/l) SD (U/l) %CV SD (U/l) %CV The LIPASE liquicolor reagent shows a good intra- and interassay precision in the low, middle, and high concentration range. 3 Linearity and Detection Limit 3.1 Linearity The linearity of LIPASE liquicolor (lot SA-115, exp date ) was controlled by employing a high concentrated pool serum successively diluted with phys. Saline. The analysed concentrations were compared with the theoretical concentrations obtained from a linear regression. Deviations of measured against theoretical concentrations are expressed in absolute and relative values. Linearity was assumed within an accepted deviation of ± 10%. As an example data measured on the AU 400 system are reported. High Pool Analytical Data Regressed Data Deviation from Regression Line Content (%) U/l U/l U/l (%) The LIPASE liquicolor test is linear up to an Lipase concentration of at least 300 U/l. Design Verification and Product Data for Lipase 2/9

3 3.2 Detection Limit The detection limit has been evaluated from a 20-fold repeat of a zero sample (saline). Lipase liquicolor reagent: [BUF] lot and [SUB] , exp date The mean value was U/l with a standard deviation of U/l. From these results the detection limit has been calculated acc. to the formula: abs. mean + 3 SD = detection limit. The detection limit resulted to: 0.32 U/l. 4 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. The determinations of each control serum have been performed with LIPASE liquicolor (lot SA-115, exp date ). The mean values have been calculated and compared with the target values of the respective control sera. CONTROL SERUM RECOVERY HUMAN fresh Control LOT Target Range Result Deviation to Fresh serum U/l U/l U/l (%) Precinorm # Precipath # C.fas # Autocal Chema # 9A Mean Comparison of Methods LIPASE liquicolor reagent (lot SA-115, esp date ) run on Olympus AU 400 has been compared against Lipase reagent Roche (lot , exp date ) measured on Olympus AU 400 analyzer. Results < 40 U/l have been excluded. Control sera as well patient samples have been employed in the comparison (N=39). 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 0.90 and 1.10, acceptance criteria ± 10%. The linear regression obtained can be described as follows: r = Y = X X mean = U/l Y mean = U/l Sample Mean Reference Mean Test No. Lipase (Roche) Lipase Design Verification and Product Data for Lipase 3/9

4 Mean Both methods showed a good agreement and no significant deviation could be observed with any specific sample. Design Verification and Product Data for Lipase 4/9

5 6 Stability, Accelerated Stress 6.1 Linearity The linearity of LIPASE liquicolor (lot SA-115, exp date ) was controlled according to the procedure already described in 3.1. The reagents were stressed 10 days at 37 C. As an example data measured on the AU 400 system are reported. High Pool Analytical Data Regressed Data Deviation from Regression Line Content (%) U/l U/l U/l (%) The test is specified to cover a linear range up to about 300 U/l as indicated in section 3.1. The above results clearly confirm that the test achieves its specification after accelerated stress. 6.2 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. The determinations of each control serum have been performed with LIPASE liquicolor (lot SA-115, exp date ). The mean values have been calculated and compared with the target values of the respective control sera. Deviation to Fresh acceptance criteria is ± 10%. Design Verification and Product Data for Lipase 5/9

6 CONTROL SERUM RECOVERY HUMAN fresh HUMAN 10d 37 C Control LOT Target Range Result Result Deviation to Fresh serum U/l U/l U/l U/l (%) Precinorm # Precipath # C.fas # Autocal Chema # 9A ,1 Mean Accelerated stress data are indicative for a real-time stability of 18 months at a storage temperature of C. 7 Real Time Stability 6.1 Recovery of Control Sera Commercially available control sera and patient sera have been employed. The control sera have been reconstituted/prepared according to the manufacturer s instructions. The determinations of each control and patient serum have been performed with LIPASE liquicolor test. The mean values obtained with fresh (lot # FA-081) and three different reagent lots (lot # SA-133, lot # SA-171, lot # SA-074) have been calculated and compared with the target values of the respective control sera and means established with fresh reagent lot. SERUM AND CONTROL RECOVERY / Lot# FA-081 Lot# SA-133 Lot# SA-115 Lot# SA-074 Real Time Fresh 18 months 22 months 24 months Target [U/l] Range [U/l] Result [U/l] Result [U/l] Result [U/l] Result [U/l] Precinorm # Precipath # Hum Asy Level # 858UN Hum Asy Level # 651UE Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient All results support the claimed stability of 18 months from production. Design Verification and Product Data for Lipase 6/9

7 8 Open Vial Stability LIPASE liquicolor open vial stability has been tested for 28 days on the AU 400 system. The calibration has been performed on the first day. 8.1 Linearity The linearity of LIPASE liquicolor was controlled according to the procedure already described in 3.1. with fresh reagent and reagent stored on AU 400 system for 28 days. Open vial stability was assumed by linearity testing within an accepted deviation from regression line of ± 10%. The results are summarised below: High Pool Analytical Data LIPASE liquicolor reagent fresh [BUF] lot and [SUB] , exp date Regressed Data Deviation from Regression Line Analytical Data LIPASE liquicolor reagent 28 d open vial [BUF] lot and [SUB] , exp date Regressed Data Deviation from Regression Content (%) U/l U/l U/l (%) U/l U/l U/l (%) Line 8.2 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. The calibration has been done on the first day of measurements. The determinations of each control serum have been performed with LIPASE liquicolor fresh and after 28 days open vial ([BUF] lot and [SUB] , exp date ). The mean value has been calculated and compared with the fresh mean of the respective control sera (accepted deviation ± 10%). Calibration stability Open vial stability CONTROL SERUM RECOVERY LIPASE liquicolor LIPASE liquicolor LIPASE liquicolor Fresh 8 days opened 28 days opened Control Lot Target [U/l] Range [U/l] Result [U/l] Result [U/l] Deviation % Result [U/l] Deviation % SERODOS SERODOS Plus Precinorm Precipath TruN TruP Sum Mean The on board stability and calibration stability of LIPASE liquicolor is confirmed for a minimum of 28 days. The calibration has been performed on the first day. The calibration stability is confirmed for a minimum of 8 days. Design Verification and Product Data for Lipase 7/9

8 9 Interferences Interference by bilirubin. hemoglobin, triglycerides and ascorbic acid have been studied by adding known amounts of the potentially interfering substance to a known sample. Recoveries have been analysed according to the method of Glick et al. (Clin.Chem ). Criterion :Recovery within ± 10 % of initial value. The results are summarised in the following table. Lipemia Triglycerides mg/dl Mean Lipase U/l Recovery % Bilirubin Bilirubin mg/dl Mean Lipase U/l Recovery % Hemoglobin Haemoglobin mg/dl Mean Lipase U/l Recovery % Ascorbic Acid Ascorbic Acid mg/dl Mean Lipase U/l Recovery % Design Verification and Product Data for Lipase 8/9

9 Interferences < 10%: Hemolytic samples up to 400 mg/dl hemoglobin by Bilirubin no interference up to 50 mg/dl by Triglycerides no interference up to 1100 mg/dl by Ascorbic acid no interference up to 50 mg/dl 10 Traceability LIPASE liquicolor calibration is performed with AUTOCAL REF and traceable to an in-house calibrator. Design Verification and Product Data for Lipase 9/9