GB/T 43733-2024 in English

National Standard of the People's Republic of China GB/T 43733—2024

Determination of Chlorogenic Acids in Plants by High Performance Liquid Chromatography

Background of the Standard and Background of Formulation

This standard was proposed and coordinated by the National Technical Committee for Standardization of Biochemical Testing (SAC/TC 387), and the main drafting units include Shenzhen Institute of Metrology and Quality Inspection, Shenzhen Agricultural Product Quality and Safety Inspection and Testing Center and other authoritative institutions. The formulation of this standard aims to unify the determination methods of chlorogenic acids in plants and ensure the accuracy and consistency of the test results.

Scope of Application

This document is applicable to the determination of 6 chlorogenic acids (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C) in samples such as roots, stems, leaves, flowers, fruits, and seeds of plants.

Technical Specifications and Operation Procedures

Sample Preparation: Solid samples need to be crushed evenly and stored in a refrigerator, while fresh samples need to be homogenized and stored in a frozen state. This step ensures the uniformity and stability of the sample.

Extraction Process: Accurately weigh the sample and add it to a 50% methanol aqueous solution, perform ultrasonic extraction for 30 minutes, and filter the sample to a constant volume before testing. This method is highly efficient and can retain the target compound to the maximum extent.

Instrument and Reagent Requirements

• High Performance Liquid Chromatograph: Equipped with UV detector.
• Electronic Balance: Sensitivity 0.1g and 0.1mg.
• Ultrasonic Oscillator and Centrifuge: Ensure extraction and separation effects.

Standard curve and result calculation

By drawing the standard curve, the relationship between peak area and concentration is used for quantitative analysis. The calculation formula is shown in formula (1):

Wherein, $X_{i}$ is the content of each component in the sample (unit: mg/kg), $c_{i}$ is the concentration of each component in the sample solution (unit: μg/mL), $V$ is the fixed volume of the sample solution (unit: mL), $f$ is the dilution factor of the test solution, and $m_{i}$ is the sample mass (unit: g).

Precision and detection limit

Under repeatability conditions, the absolute difference between two independent determination results should not exceed 10% of the arithmetic mean. The detection limit of the method is 1.5 mg/kg, and the quantification limit is 5 mg/kg.

Implementation suggestions

In practical applications, it is recommended to give priority to high performance liquid chromatography for detection, and ensure the calibration and maintenance of reagents and instruments. At the same time, adjust the extraction conditions according to the sample type to obtain the best detection effect.

Appendix: Comparison of standard frameworks