Peak table column components


Introduction

This section contains a list of peak parameters with explanations and calculation formulae when applicable.


Peak parameters - illustration

The diagram below illustrates the peak parameters. See the parameter list below for explanations.


Peak parameter descriptions

The list below contains descriptions of the peak parameters.

Parameter

Description

Amount

Values calculated by the Analysis module. (Only available if the Quantitation module is installed.)

Area

Calculated as the area between the curve and baseline, between the peak start and peak end, time or volume base. (Gray area in the diagram above.)

Asymmetry

Peak asymmetry (indicator of column packing). See definition below this table.

Baseline height

Baseline amplitude at peak start, peak maximum and peak end. (A, F and G in the diagram above.)

Capacity factor

The capacity factor will only be calculated when the chromatogram is in volume base. The total liquid volume, Vt, must be entered in the Integrate dialog box for this parameter to be calculated. See definition below this table.

Concentration

Values calculated by the Analysis module. (Only available if the Quantitation module is installed.)

Fraction tube id

Fraction number at peak start, peak maximum and peak end.

Height

Maximum amplitude above the baseline. (C-F in the diagram above)

Kav

Gel phase distribution constant in gel filtration. Kav will only be calculated when a gel filtration column was used and when the chromatogram is in volume base. The void volume, V0, must be entered in the Integrate dialog box for this parameter to be calculated. See definition below this table.

Molecular size

Values calculated by the Analysis module. (Only available if the Quantitation module is installed.)

Plate height (HETP)

Height equivalent to theoretical plate and plates/meter. The column height must be entered in the Integrate dialog box for this parameter to be calculated. See definition below this table.

Peak endpoint heights

Amplitude above the baseline at left (A in the diagram above) and right peak limits (E-G in the diagram above).

Peak endpoint retention

Retention value at peak start and peak end, time or volume base. (A and G in the diagram above.)

Peak name

Name of the peak.

Percent of total area

Peak area as a percent of the total area under the curve above the baseline. Time or volume base.

Note: This value can differ in time and volume base if the flow rate is not constant throughout the method.

Percent of total peak area

Peak area as a percent of the sum of all integrated peaks.

Note: This value can differ in time and volume base if the flow rate is not constant throughout the method.

Resolution

Peak resolution. See definition below this table.

Retention

Retention at the peak maximum, time or volume base. (C in the diagram above.)

Sigma

Standard deviation for a Gaussian-shaped peak. See definition below this table.

Type of peak limits

Identifies the criteria for peak start and peak end as either the baseline intersection or dropline to the baseline or skim line.

Width

Difference in retention between the peak end and peak start, time or volume base. (G-A in the diagram above.)

Width at half height

Calculated by taking the maximum height of the peak above the baseline, then determining the peak width at half this value above the baseline. Time or volume base. (B-D in the diagram above, where BD bisects CF.)

Note: In the Options dialog box in the UNICORN Manager you can select if negative retentions should be displayed or not. The default selection is that negative retention is not displayed.


Sigma formula

The formula below is used to calculate Sigma.

Where:

  • n is the number of data points.

  • x is the volume or time value.

  • x ymax is the volume or time value at the maximum amplitude value.

  • Apeak is the area of the peak.

Note: The peak width for a Gaussian peak is (4 x Sigma).


Peak resolution algorithms

The peak resolution is calculated with one of the following three algorithms:

  1. (VR2 - VR1) / ((Wb2 + Wb1) / 2)

  2. (VR2 - VR1) / ((Sigma2 + Sigma1) x 2)

  3. ((VR2 - VR1) / (2 x (Wh2 + Wh1))) / 2.354

Where:

  • VR1, Wb1, Sigma1 and Wh1 are the retention, width, Sigma and width at half height of the previous peak.

  • VR2, Wb2, Sigma2 and Wh2 are the retention, width, Sigma and width at half height of the current peak.

Note: The Resolution algorithm variable in the Options dialog box in the UNICORN Manager determines which of the three algorithms is used. If this variable has the value 1, 2 or 3, then the algorithm with the corresponding number in the list above is used. The default value is 3.


How to change the peak resolution algorithm

The table below describes how to change the peak resolution algorithm in the UNICORN Manager.

Step

Action

1

  • Choose the Administration:Options... menu item.

Result: The Options dialog box opens.

2

  • Select the desired algorithm number described as described in Peak resolution algorithms above, in the Resolution algorithm droplist.

  • Click OK.

Result: The dialog box closes and the peak resolution algorithm is changed.

Note: You must repeat the peak integrations after the change to update the values based on the new algorithm.


Capacity factor formula

The formula below is used to calculate the Capacity factor.

Where:

  • VR = retention volume.

  • Vt = total liquid volume.


Kav formula

The formula below is used to calculate Kav.

Where:

  • VR = retention volume.

  • V0 = void volume.

  • VC = column volume.


Asymmetry formula

The formula below is used to calculate the Asymmetry.

Asymmetry = B / A

Where:

  • A is a partial peak width, measured at a percentage of the peak height, for the leading part of the peak.

  • B is a partial peak width, measured at a percentage of the peak height, for the tailing part of the peak.

How to change the Asymmetry Ratio

The Asymmetry Ratio is selected in the Options dialog box in the UNICORN Manager. The table below describes how to select a value:

Step

Action

1

  • Choose the Administration:Options... menu item.

Result: The Options dialog box opens.

2

  • Type a ratio value in the Asymmetry Ratio at text box.

  • Click OK.

Result: The ratio value is changed and the dialog box closes.

Note: You must repeat the peak integrations after the change to update the values based on the new asymmetry ratio. The default ratio is 10%.


HETP formula

The formula below is used to calculate the HETP value.

HETP = L/N

N = 5.54 x (VR/wh)2 assuming a Gaussian peak.

Where:

  • N = no. of theoretical plates.

  • L = bed height in cm.

  • VR = peak retention (elution) volume or time.

  • wh = peak width at half height expressed in the same units as VR.


2005-06-15