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` The Working a vera Calibration Factor or Calibration Curve ga must be verified on each corking day by the introduction of one or more calibration standards . The frequency of ' verification is detector dependent, and varies from once per day to an average of once every five samples . o If the response of any analyte varies Tran the predicted response by more than +!- 15X, a new calibration curve must be prepared for that analyte . (R1-R2) o The Percent Difference - ------ x 100, where R1 R1 the CF from the first analysis, and R2 - the CF from the second analysis . K o Internal Standard Calibration Procedures ror each analyte, or group of analytes, fiue concentration leuels of standards are prepared by adding aliquots of one or more stock standards to uolumetric flasks . In addition, a known and constant amount of one or nore internal Standards CIS,$)is added to each uolumetric flask and they are then diluted to volume with an appropriate solvent . one of the standards should be at a concentration near the method detection limit . The other concentrations should define the working range of the systen. Each of the calibration standards is introduced into the chromatography system using the technique to be used for introduction of actual samples Cie . 2- to 5-ul liquid injections, purge M trap, etc.) . Both a Calibration Curue, and a series of Calibration Factors CCr,$) at each standard concentration, is calculated for each analyte . o The Calibration Curve is a plot of amount introduced us . the relative detector response (RR) . response-of-analytes o The RR response of IS mass introduced a The Cf a ----------------------- (for nultiresponse y total relative response compounds use the total area of all peaks used for quantitation) . o If the percent relative standard deviation (XRSD) between the Cr's is less than 20X over the working range, linearity through the origin can be assumed and an average Cf can be used for quantitation. 19 d