Summary
A new equation which describes the interdependence of instrument band spreading, injection volume and input profile is proposed and experimentally verified in two modes: 1. without a column; 2. with a column.
It is shown that under equivalent conditions both modes are in excellent agreement. The method of calculation of the variance of a response function appeared to be of utmost importance in correctly interpreting the experimentally observed interdependence of instrumental band spreading and injection characteristics.
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Abbreviations
- a0, a1, a2 :
-
Coefficients of the parabolic equation applied in the curve fitting of the data pairs
- b0, b1 :
-
Coefficients of the linear equation applied in the curve fitting of the data pairs
- D:
-
Factor depending on input profile and calculation method
- Dm :
-
Diffusion coefficient of a solute in the mobile phase
- Dp :
-
Average diffusion coefficient of a solute in a particle of the stationary phase
- dp :
-
Average particle diameter of the stationary phase
- F:
-
Volume flow rate
- k′:
-
Capacity factor
- M0 :
-
Zeroth moment
- M1 :
-
First normalized moment\(\int\limits_0^\infty {{{th(t)dt} \mathord{\left/ {\vphantom {{th(t)dt} {M_0 }}} \right. \kern-\nulldelimiterspace} {M_0 }}} \)
- M2 :
-
Second normalized central moment\(\int\limits_0^\infty {{{(t - M_1 )^2 h(t)dt} \mathord{\left/ {\vphantom {{(t - M_1 )^2 h(t)dt} {M_0 }}} \right. \kern-\nulldelimiterspace} {M_0 }}} \)
- Ncol :
-
Number of theoretical plates generated by the column
- r:
-
Regression coefficient of fitted curves
- \(\hat S_{yx}^{n - 2} \) :
-
Modified standard error of estimate for (n−2) degrees of freedom
- tR :
-
Retention time of eluted component
- u0 :
-
Linear velocity of an unretained component
- Vcol :
-
Column volume
- Vinj :
-
Volume of injected sample
- \(\bar y^{obs} \) :
-
Mean value of the dependent variable for different types of regression
- \(\sigma _{v(ext)}^2 \) :
-
Volume variance of concentration profile due to injection volume, injection device, tubing and detector (external broadening)
- \(\sigma _{v(inj)}^2 \) :
-
Volume variance of concentration profile due to injection volume
- \(\sigma _{v(col)}^2 \) :
-
Volume variance of the impulse response of the column
- \(\sigma _{v(0)}^2 \) :
-
Equal to\(\sigma _{v(ext)}^2 \) for Vinj→0, impulse response of the instrument
- \(\sigma _{v( )}^2 \) :
-
Time variances of above mentioned concentration profiles
- ∈t :
-
Total porosity of column bed
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Presented as part of a poster at the Vth International Symposium on Column Liquid Chromatography, Avignon (France), 11–15 May 1981.
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Lauer, H.H., Rozing, G.P. The selection of optimum conditions in HPLC I. The determination of external band spreading in LC instruments. Chromatographia 14, 641–647 (1981). https://doi.org/10.1007/BF02291104
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DOI: https://doi.org/10.1007/BF02291104