Level 5 specify the amount of the component in each level of check standard When you enter an amount, the Total Levels field is

Component Data

Clicking the Detail... button or the number button next to each component on the Component Table opens the Component Data dialog box. The dialog box consists of three tab pages:

• Plot displays a plot of response vs. amount for any component of the current Method.

• Replicates displays the raw area or height calibration data for up to 10 replicate calibration injections, at each calibration level. • Groups lets you classify components and/or unknown peaks into

groups to get reports of compound classes.

Component Data Plot Tab Page

Component selects the component plot to display.

Curve Fit Type selects the curve fit to use to calculate the plot. When a new curve fit is selected the plot is updated immediately.

Calibrate By selects whether the Area or the Height is used in the calibration plot.

NOTE The Curve Fit Type, Origin, and Calibrate By options are the same as those set in the Component Table - Calibration Tab Page. You can test new setting here, and then either select Apply or OK to save the new settings, or select Cancel to keep the original settings.

Display controls the type of information included in the plot. Check or clear the individual Display options check boxes below the plot, or point to the plot and click the right mouse button to select options from a pop-up menu.

Included Pts displays the amount, response data points for each calibration level.

Curve draws the selected Curve Fit Type.

Error Bars displays a bar between replicate values of one standard deviation or more from the average value. Narrower bars indicate closer replicate values, wider bars indicate greater deviation in replicate values.

Rejected Pts displays an X at the location of each data point not used in the plot.

Residuals displays the plot of residual versus amount. A residual is the difference between the actual measured Y value and the Y value that is generated by the regression calibration for a given X value. For the calibration plot, the residual is the difference between the actual response of a calibration standard and the response that would be expected based on a regression curve. The residual axis is on the right side of the plot. For each point, a circle indicates the residual value, with a vertical line indicating the difference from the ideal residual value of zero.

Confidence Intervals draws curves showing the confidence intervals for the plot. This option is available only for External Standard Linear curve fits with two or more levels. The confidence intervals are calculated using a t-test at 95% confidence.

NOTE If there are only small differences between replicate data points, or if the residuals for the data points are small, the confidence curves may be indistinguishable from the calibration curve.

Print... opens a Print dialog box for printing the plot. You can select whether to print all components or only the current component.

Replicates displays the data values (area or height) by either replicate or level.

View selects whether to list replicate data by Level or Replicate. When Level is selected, the table displays all replicate data values for the selected level. Rows are labeled R1, R2, . . . Unused (empty) Levels are not included in the list.

If Replicate is selected, the table displays a data value for each level of the selected replicate calibration injection. Rows are labeled L1, L2, . . . The Use column, indicates which values to use in the calibration plot. The values can be manually accepted or rejected, or you can set up automatic rejection of outlying data points from the Calibration Parameters dialog box (see Section 5.8.4).

A Comment (up to 256 characters) is required for rejected values. For accepted values, entering a comment is optional.

Calculator lets you calculate an amount from a response, or a response from an amount, for the current component.

To calculate an amount from a response, enter the Response value and then click in the Amount box. A new amount is calculated based on the entered response. To calculate a response from an amount, enter the Amount and click in the Response box.

The following values, used to calculate component amount, are determined automatically by the Method Editor and cannot be edited.

K0 indicates the Y-intercept of the calibration curve.

K1 is the coefficient for the first-degree variable. When the fit type is linear, K1 indicates the slope of the calibration curve for the selected calibration level.

K2 is the coefficient for the second-degree variable in a quadratic fit. K3 is the coefficient for the third-degree variable in a cubic fit. For example, to determine the cubic fit of peak area calibration, the equation is:

Component Data Replicates Tab Page

Component selects the component for which to display the replicate data. View selects whether to display the Area or Height calibration data for the selected component.

The spreadsheet shows the area or height values for the selected

component. Data from up to 10 replicates and 32 levels can be displayed. Rejected calibration points are shown in strike-out font.

The equation for the calibration curve fit used to calculate the component amount is displayed at the bottom of the Replicates page. You can select the curve fit from either the Calibration tab page of the Component Table dialog box or from the Plot tab page of the Component Data dialog box. The r2 value (Coefficient of Determination) for the component is shown at the bottom of the Replicates page. The ideal value for this coefficient is 1.0.

Component Data Groups Tab Page

New groups are created in the Add Component Groups dialog box. Use one of the following methods to open the dialog box:

• If there are no groups currently defined, select a component name from the Add Ungrouped Component list and then click the Add button, or select the Add Unknown Peaks option.

• If groups currently exist, double-click the Groups drop-down and then click the Add button.

A group may contain any number of components, and as many as five different time ranges. Enter a Group Name here and click on Add to enter the data in the Component Table. Repeat for each component group.

When you finish adding component groups, click on Exit to return to the Component Groups tab page.

Group indicates the name and number of the component group being edited. The list box directly below displays the group’s components; the number preceding each component is its retention time. To see information for a different component group, click the cursor on the down-arrow beside the group name, then click on the group from the list of names now

displayed. Double-clicking on an item in the Group list box transfers it to the Add Ungrouped Component list.

Add Ungrouped Component displays the names and retention times of all ungrouped components. There are two ways to add an ungrouped

component to the group being edited (or to remove a component from the group being edited): double-click on the component name, or click first on the component name and then on the Add (or Remove) command button. Add Unknown Peaks adds unknown peaks that appear within a particular time range to the component group being edited. Enter the beginning and end of the time range (in minutes) in the edit boxes here and then click on the Add command button. The word “unknowns,” is displayed after the time range in the list of groups.

Define Group As a Component adds a group to the Method’s Component Table. The group to be added must be made up of only unknown peaks (see Add Unknown Peaks above). The Component Table will list the group name as a single component; the retention time for the grouped component will be the average of the time ranges of the unknown peaks in the group. When a calibration standard for the group is run, the total area of all peaks in the group is entered into the Replicates table in the Component Table. The calibration curve is calculated with the group treated as a single

component. When the unknown sample is run, PeakNet determines the total area of the unknown peaks in the group and determines the amount of that group in the sample based on the calibration curve.

Add removes the highlighted component in the Add Ungrouped Component window and transfers it to the group being edited or, if the name of the group being edited is highlighted, clicking on Add opens the Add Component Groups dialog box.

Remove transfers a highlighted component name from the group list to the Add Ungrouped Component window. If the Group name is highlighted, clicking on Remove deletes the entire group.

5.9