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Column Efficiency Testing

The document outlines 7 different calculations used in chromatography: 1) Calculation of theoretical plates using the half-height and USP methods 2) Calculation of peak asymmetry factor and tailing factor 3) Calculation of height equivalent to a theoretical plate (HETP) 4) Calculation of reduced plate height 5) Calculation of chromatographic resolution It provides the equations for calculating various chromatography metrics like number of theoretical plates, asymmetry, HETP, reduced plate height, and resolution.

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908 views3 pages

Column Efficiency Testing

The document outlines 7 different calculations used in chromatography: 1) Calculation of theoretical plates using the half-height and USP methods 2) Calculation of peak asymmetry factor and tailing factor 3) Calculation of height equivalent to a theoretical plate (HETP) 4) Calculation of reduced plate height 5) Calculation of chromatographic resolution It provides the equations for calculating various chromatography metrics like number of theoretical plates, asymmetry, HETP, reduced plate height, and resolution.

Uploaded by

vamsi
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

1.

Calculation of the number of Theoretical Plates (half-height method, used by Tosoh)

Where: N = Number of theoretical plates Ve = elution volume or retention time (mL, sec, or cm) h = peak height w1/2 = width of the peak at half peak height (mL, sec, or cm)

2. Calculation of the number of Theoretical Plates (USP method)

Where: N = Number of theoretical plates Ve = elution volume, retention time or retention distance (mL, sec, or cm) h = peak height wb = width of the peak at the base line (mL, sec, or cm)

3. Calculation of peak Asymmetry Factor (used by Tosoh)

Where: As = peak asymmetry factor b = distance from the point at peak midpoint to the trailing edge (measured at 10% of peak height) a = distance from the leading edge of peak to the midpoint (measured at 10% of peak height)

4. Calculation of Tailing Factor (USP method)

Where: T = tailing factor (measured at 5% of peak height) b = distance from the point at peak midpoint to the trailing edge a = distance from the leading edge of the peak to the midpoint

5. Calculation of the Height Equivalent to a Theoretical Plate (HETP)

Where: H = Height equivalent of a theoretical plate L = Length of the column N = Number of theoretical plates Dimensions: when using HPLC or UHPLC columns, H is usually expressed in m.

6. Calculation of Reduced Plate Height (h)


Giddings introduced dimensionless parameters for H and also for the linear velocity. Dimensionless parameters allow the direct comparison of the efficiency of two or more columns packed with different particle size packing materials. According to the theory, a well packed column should have a reduced plate height (h) in the range of 2-3 at a reduced velocity (v) of about 3. Here we only provide the formula for h.

Where: h = reduced plate height (sometimes referred to as the number of band widths) H = height equivalent of a theoretical plate (m) dp = mean particle size (m)

7. Calculation of chromatographic Resolution

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