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HPLC Mobile Phase (Gradient)

HPLC Mobile Phase (Gradient) Calculator | PharmCal Professional

HPLC Mobile Phase (Gradient)

Calculate Quaternary (A, B, C, D) solvent volumes based on gradient table.

1. BASIC PARAMETERS
mL/min
min
Inj.
%
2. GRADIENT TABLE PROGRAM
Time (min) % A % B % C % D
PREPARATION VOLUMES (w/ Overage)
Total Mobile Phase: 0 mL
Solvent A: 0 mL
Solvent B: 0 mL
Solvent C: 0 mL
Solvent D: 0 mL
*Volumes based on integrated gradient profile.
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Understanding Gradient Mobile Phase Calculation

In complex HPLC separations, an isocratic (constant) solvent composition is often insufficient. Gradient Elution involves changing the mobile phase composition over time to elute compounds with varying polarities. This tool calculates the exact volume needed for each solvent channel (A, B, C, D) based on your specific gradient program.

Step 1: Basic Parameters

Enter the Flow Rate (mL/min), total Runtime per injection (including column equilibration), and the total Number of Injections.

Step 2: Define Gradient Table

Input your method's gradient table row by row. Enter the Time and the percentage of each solvent (A/B/C/D).
Note: The sum of A+B+C+D must equal 100% for each row.

Step 3: Calculate

Click "Calculate". The tool performs a mathematical integration (Trapezoidal Rule) under the gradient curve to determine the Average % Composition of each solvent over the course of the run. It then applies this average to the total required volume (plus your specified overage).

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