Flow Cell Path Length

Lambert-Beer’s law shows a linear relationship between the flow cell path length and absorbance.

where

T is the transmission, defined as the quotient of the intensity of the transmitted light I divided by the intensity of the incident light, I₀,

ε is the extinction coefficient, which is a characteristic of a given substance under a precisely-defined set of conditions of wavelength, solvent, temperature and other parameters,

C [mol/L] is the concentration of the absorbing species, and

d [m] is the path length of the cell used for the measurement.

Therefore, flow cells with longer path lengths yield higher signals. Although noise usually increases little with increasing path length, there is a gain in signal-to-noise ratio. For example, in Influence of Cell Path Length on Signal Height the noise increased by less than 10 % but a 70 % increase in signal intensity was achieved by increasing the path length from 6 – 10 mm.

When increasing the path length, the cell volume usually increases — in our example from 5 – 13 µL. Typically, this causes more peak dispersion. As Influence of Cell Path Length on Signal Height demonstrates, this did not affect the resolution in the gradient separation in our example.

As a rule-of-thumb the flow cell volume should be about 1/3 of the peak volume at half height. To determine the volume of your peaks, take the peak width as reported in the integration results multiply it by the flow rate and divide it by 3).

Traditionally LC analysis with UV detectors is based on comparing measurements with internal or external standards. To check photometric accuracy of the detector it is necessary to have more precise information on path lengths of the flow cells.

The correct response is:

expected response * correction factor

Please find below the details of the flow cells:

base-id: 3358186379

id: 3358186379