Use of a ISO template for quantitative schemes : the issues. Annette Thomas
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1 Use of a ISO template for quantitative schemes : the issues Annette Thomas
2 ISO and homogeneity and Stability ISO requires that an EQA provider demonstrates sufficient homogeneity and stability of the material using valid statistical methods. The Standard refers the reader to ISO Statistical methods for use in proficiency testing by interlaboratory comparisons and the IUPAC International Harmonized Protocol. The procedure described in ISO Annex B is most suited for large quantitative schemes with little guidance provided for Schemes where there are insufficient number of samples for statistical validity, or where split sample analysis is inappropriate. There is little or no guidance as to the design of suitable homogeneity and stability testing protocols for qualitative analytes.
3 Issue 1 wide method CV The standard statistical approach for homogeneity and stability includes calculation of the general average, the within sample and between sample standard deviation. The samples are considered to be adequately homogeneous if the between sample sd is 0.3* Standard deviation for proficiency assessment (SDPT) t represents the sample (t = 1, 2..., g) k represents the test portion (k = 1, 2) Calculate sample averages as: and the between-test-portion ranges as: Calculate the general average: the standard deviation of sample averages: and the within-samples standard deviation: Calculate the between-samples standard deviation as: Homogeneous if
4 between-samples standard deviation = Issue 1 wide method If SD PT = 0.5 then SD allow = 0.3x SD PT = 0.15 and If S x = 0.5 and S w = 0.8 CV Then S s = ( /2) = ( ) = If S w > S x look at analytical variation of method If S s is negative can t use standard equation use alternative SD allow which takes into account within sample SD.
5 Calculate SD allow. Calculate F1 and F2 are from standard statistical tables, Issue 1 wide method CV If ss > c then there is evidence that the batch of material is not sufficiently homogeneous For 10 samples, F1 = 1.88 and F2 = 1.01 c = 1.88 x SD 2 allow x(within sample SD) 2 c = 1.88 x x c = = 0.69 c = 0.83
6 If Within sample SD > Sample Av SD Analyte: Glucose Analyte: Glucose Analyte: Glucose Batch no: (please enter) Sample Sample Sample Result a Result b Sample Ave Result a Result b Sample Ave Result a Result b Sample Ave Sample Sample Sample Sample Sample Sample Sample Sample Sample Sample General Average Sum d Within Sample SD (SD diff ) Compare d 2 and remove data points (use Cochran s test if needed. SD of sample ave Between sample SD #NUM! enter WEQAS SD for this level (calculated from precision profile & general average or POCT criteria (in%)) *WEQAS SD SD 2 allow C DATE SAMPLES SENT TO ANALYSER: Between sample SD (1) must be < 0.3*WEQAS SD (2) or c (3) DATE OF ANALYSIS SD 2 allow = (0.3 *WEQAS SD) 2 c = 1.88*SD 2 allow *(within sample SD) 2
7 Issue 2-Where the SD PT is small Impossible to achieve 0.3 x SD Tight performance criteria based on Clinical allowable limits. Challenging samples near the analytical limits of the assay. In these cases the criterion should be expanded to allow for the sampling errors.
8 Issue 3 Random v Systematic ISO Guide 34 - systematic sampling in a continuous process may be a better way to detect inhomogeneity rather than random sampling. Evaluation of the data should include investigation of a trend (or drift) in analysis of the homogeneity measurements, or a trend during the dispensing or production processes.
9 Random sampling approach Systematic sampling approach Analyte: Glucose Analyte: Glucose Batch no: (please enter) Sample Sample Analysis sequence of all samples should be random for both approaches to minimise affects of analyser drift Result a Result b Sample Ave Result a Result b Sample Ave Sample Sample Sample Sample Sample Sample Sample Sample Sample Sample General Average Sum d Within Sample SD (SD diff ) SD of sample ave Between sample SD enter SD for this level *WEQAS SD SD 2 allow C DATE SAMPLES SENT TO ANALYSER: Between sample SD (1) must be < 0.3*WEQAS SD (2) or DATE OF ANALYSIS SD 2 allow = (0.3 *WEQAS SD) 2
10 Trend Analysis - Random sampling Systematic sampling provides greater information than random sampling Bias to General Av Pair 1 Pair Trend Analysis - Systematic sampling Sample pair Bias to General Av Pair 1 Pair Sample pair
11 Issue 4 Small sample nos. Use fewer points in ISO template and use range instead of SD If n = 2,3 then (x high x low ) = ss If n =4,5,6 then (x high x low )/1.5 = ss If n =7,8,9 then (x high -x low )/2 = ss Alternative approach is to assess the observed SD PT against the predicted Standard deviation calculated from the Precision profile derived from previous rounds.
12 Use 5 instead of 10 pairs Batch no: (please enter) Analyte: Glucose Analyte: Glucose Result a Result b Sample Ave Result a Result b Sample Ave Sample Sample Sample Sample Sample Sample 6 Sample 7 Sample 8 Sample 9 Sample 10 General Average Sum d Within Sample SD (SD diff ) SD of sample ave Between sample SD enter WEQAS SD for this level (calculated from precision profile & general average or POCT criteria (in%)) *WEQAS SD SD 2 allow C Sample Sample DATE SAMPLES SENT TO ANALYSER: Between sample SD (1) must be < 0.3*WEQAS SD (2) or DATE OF ANALYSIS SD 2 allo w = (0.3 *WEQAS SD) 2
13 Compare against previous SD Precision Profile Glucose SD Glucose Concentration (mmol/l) Jan '99 - Mar '00 Sep '04 - Jan '05 Feb '05 - Jul '05 Jul '05-Nov'05 Dec '05 - Apr '06 May '06 - Sep '06 Oct '06 - Feb '07 Marc '07 - Jul '07 Aug '07 - Mar '08 Biological goals Poly. ( ) Linear (Biological goals) y = x x R 2 = y = x x R 2 =
14 Assessment criterion for qualitative Schemes Not covered in ISO Split sampling procedure could be followed as for Quantitative analytes. The samples could be considered homogeneous if all samples (100% responses) tested gave the same result. If this is not the case compare the variability of results from the within vial results and the between vial results. i.e. calculate false negatives/ true positive % for within vial calculate false negatives / true positive % for between vials false negatives/ true positive % for between vials false positive / true positive % for within vial.
15 Conclusion Homogeneity and stability testing requires a more pragmatic approach than the procedure described in ISO 13528:2005
GAMINGRE 8/1/ of 7
FYE 09/30/92 JULY 92 0.00 254,550.00 0.00 0 0 0 0 0 0 0 0 0 254,550.00 0.00 0.00 0.00 0.00 254,550.00 AUG 10,616,710.31 5,299.95 845,656.83 84,565.68 61,084.86 23,480.82 339,734.73 135,893.89 67,946.95
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