GB/T 43733-2024
Determination of chlorogenic acids in plants by high performance liquid chromatography (English Version)

Standard No.
GB/T 43733-2024
Language
Chinese, Available in English version
Release Date
2024
Published By
General Administration of Quality Supervision, Inspection and Quarantine of the People‘s Republic of China  CN  /  GB
Latest
GB/T 43733-2024
 

Introduction

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.

Standard content GB/T 43733—2024 Comparative analysis
Determination method High performance liquid chromatography (HPLC) Compared with traditional spectrophotometry, the sensitivity and accuracy are significantly improved.
Detection wavelength 327 nm The optimized wavelength selection is specially designed for chlorogenic acid substances.
Extraction solvent 50% methanol aqueous solution Improve extraction efficiency and reduce impurity interference.

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):

$$ \quad\boldsymbol{X}_{i}=\frac{\boldsymbol{c}_{i}\times\boldsymbol{V}\times\boldsymbol{f}}{m_{i}} $$

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

Standard dimensions GB/T 43733—2024 Comparative advantages
Detection range 6 chlorogenic acids Covering the main plant active ingredients.
Detection sensitivity Detection limit 1.5 mg/kg Meet the needs of trace analysis.

GB/T 43733-2024 Referenced Document

  • GB/T 6682 Water for analytical laboratory use.Specification and test methods

GB/T 43733-2024 history

  • 2024 GB/T 43733-2024 Determination of chlorogenic acids in plants by high performance liquid chromatography
Determination of chlorogenic acids in plants by high performance liquid chromatography

Topics on standards and specifications

Standard and Specification

DIN EN 12137:1997 Fruit and vegetable juices - Determination of tartaric acid in grape juices - Method by high performance liquid chromatography; German version EN 12137:1997 BS EN 12137:1998 Fruit and vegetable juices - Determination of tartaric acid in grape juices - Method by high performance liquid chromatography EN 12137:1997 Fruit and vegetable juices - Determination of tartaric acid in grape juices - Method by high performance liquid chromatography EN 14176:2003 Foodstuffs Determination of domoic acid in mussels by HPLC ISO 22863-12:2022 Fireworks — Test methods for determination of specific chemical substances — Part 12: Picrates and picric acid by high performance liquid chromatography DS/EN 12137:1998 Fruit and vegetable juices - Determination of tartaric acid in grape juices - Method by high performance liquid chromatography KS M 2964-2010 Test method for determination of fatty acid methyl ester(FAME) in automotive diesel and biodiesel blends by high performance liquid chromatography(HPLC UNI 22630:2000 Seed and vegetable oils and derived products - Vegetable proteins - Determination of tryptophan aminoacid content by high performance liquid chromatography BS ISO 22863-12:2022 Fireworks. Test methods for determination of specific chemical substances - Picrates and picric acid by high performance liquid chromatography ISO 23190:2021 Traditional Chinese medicine — Determination of aristolochic acids in natural products by high-performance liquid chromatography (HPLC



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