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A simple method for HPLC retention time prediction: linear calibration using two reference substances.

Lei Sun^1,2, Hong-Yu Jin¹, Run-Tao Tian³

¹National Institutes for Food and Drug Control, No. 2 Tiantan Xili, Dongcheng District, Beijing, 100050 People's Republic of China.

Chinese Medicine

|June 24, 2017

Summary

This summary is machine-generated.

A new linear calibration method using two reference substances (LCTRS) accurately predicts retention times in High-Performance Liquid Chromatography (HPLC). This method is more robust and cost-effective than the relative retention (RR) method for pharmaceutical analysis.

Keywords:

Linear calibration using two reference substances Multi-component analysis RP-HPLC Relative retention Retention time Traditional Chinese medicines

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Area of Science:

Analytical Chemistry
Chromatography
Pharmaceutical Analysis

Background:

Accurate identification of chromatographic peaks in pharmaceutical analysis requires reference substances, which are often costly.
The relative retention (RR) method is used for unavailable reference substances but suffers from poor reproducibility across different High-Performance Liquid Chromatography (HPLC) columns.
Existing methods for predicting retention time (t_R) can be inaccurate, necessitating a more reliable approach.

Purpose of the Study:

To develop a simple and accurate method for predicting retention times (t_R) in HPLC.
To improve the reproducibility of chromatographic peak identification in pharmaceutical quality control.
To reduce the reliance on and cost associated with obtaining numerous reference substances.

Main Methods:

A novel method, linear calibration using two reference substances (LCTRS), was developed based on HPLC thermodynamic theory.
The LCTRS method involves a two-point prediction procedure, validation via multiple-point regression, and sequential matching.
Retention times (t_R) are calculated using standard retention times and a determined linear relationship.

Main Results:

The LCTRS method was successfully validated across 30 different HPLC columns using two distinct medicines.
The study demonstrated the method's applicability in real-world pharmaceutical samples.

Conclusions:

The LCTRS method is a simple, accurate, and robust alternative to the RR method for predicting retention times in HPLC.
This method enhances the reproducibility of quality standards across different laboratories.
The LCTRS method significantly lowers the cost associated with reference substances in pharmaceutical analysis.