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Home / Applications / High Speed Separation of Catechins by X-LC

  • Industry

  • Technique

High Speed Separation of Catechins by X-LC

By Miyuki Kanno

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January 5, 2024

Introduction

The main components of catechins in green tea include epicatechin (EC), epigallocatechin (EGC, hydroxy derivative of EC), epicatechin gallate (ECg, gallic acid ester of EC), and epigallocatechin gallate (EGCg, gallic acid ester of EGC).

We examined the applicability of X-LC for ultra-high speed analysis of 7 components: the 4 catechins listed above, catechin(C), catechin gallate(Cg), and caffeine.

Experimental

Jasco XLC-3000
Jasco X-LC system for high speed catechins separation

The system utilized in this experiment was a JASCO X-LC system consisting of two 3185-PU pumps, a 3080-DG degasser, a 3180-MX mixing unit, a 3067-CO column oven, a 3070-UV UV/Vis detector, a 3059-AS autosampler, and ChromNAV chromatography data system.

A commercially-available tea beverage was filtered with 0.2 µm membrane filter. A portion of the filtrate was injected.

Keywords

520015X

Results

Figure 1 shows an X-LC chromatogram of a standard mixture of 7 catechins. These components were clearly separated within 5 minutes by using a gradient elution method.

Figure 1. X-LC chromatogram of standard mixture
Peaks: 1=Epigallocatechin (500 µg/mL), 2=Caffeine (10 µg/mL), 3=Catechin (100 µg/mL), 4=Epicatechin (100 µg/mL), 5=Epigallocatechin gallate (25 µg/mL), 6=Epicatechin gallate (20 µg/mL), 7=Catechin gallate (100 µg/mL) Conditions: Column = X-PressPak AQ-C18W (2 mmID x 50 mmL, 2 µm), Column temperature = 30°C – Mobile phase: A = 0.2% phosphoric acid / acetonitrile (90/10), B = 0.2% phosphoric acid /acetonitrile (50/50), 0 min = A/B(100/0) – 0.5 min = A/B(100/0) – 4.0 min = A/B(70/30) – 5.0 min = A/B(70/30) – 5.05 min = A/B(100/0) – Flow late = 0.7 mL/min, Detection wavelength = 280 nm, Injection volume = 1 µL.

Figure 2 shows an X-LC chromatogram of the tea sample. Each component is well separated from unknown peaks.

Figure 2. X-LC chromatogram of commercially-available tea beverage
Conditions: the same as in the Figure 1.
This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

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About the Author

Miyuki Kanno

JASCO Application Note

High Speed Separation of Catechins by X-LC

Introduction

The main components of catechins in green tea include epicatechin (EC), epigallocatechin (EGC, hydroxy derivative of EC), epicatechin gallate (ECg, gallic acid ester of EC), and epigallocatechin gallate (EGCg, gallic acid ester of EGC).

We examined the applicability of X-LC for ultra-high speed analysis of 7 components: the 4 catechins listed above, catechin(C), catechin gallate(Cg), and caffeine.

Experimental

Jasco XLC-3000
Jasco X-LC system for high speed catechins separation

The system utilized in this experiment was a JASCO X-LC system consisting of two 3185-PU pumps, a 3080-DG degasser, a 3180-MX mixing unit, a 3067-CO column oven, a 3070-UV UV/Vis detector, a 3059-AS autosampler, and ChromNAV chromatography data system.

A commercially-available tea beverage was filtered with 0.2 µm membrane filter. A portion of the filtrate was injected.

Results

Figure 1 shows an X-LC chromatogram of a standard mixture of 7 catechins. These components were clearly separated within 5 minutes by using a gradient elution method.

Figure 1. X-LC chromatogram of standard mixture
Peaks: 1=Epigallocatechin (500 µg/mL), 2=Caffeine (10 µg/mL), 3=Catechin (100 µg/mL), 4=Epicatechin (100 µg/mL), 5=Epigallocatechin gallate (25 µg/mL), 6=Epicatechin gallate (20 µg/mL), 7=Catechin gallate (100 µg/mL) Conditions: Column = X-PressPak AQ-C18W (2 mmID x 50 mmL, 2 µm), Column temperature = 30°C – Mobile phase: A = 0.2% phosphoric acid / acetonitrile (90/10), B = 0.2% phosphoric acid /acetonitrile (50/50), 0 min = A/B(100/0) – 0.5 min = A/B(100/0) – 4.0 min = A/B(70/30) – 5.0 min = A/B(70/30) – 5.05 min = A/B(100/0) – Flow late = 0.7 mL/min, Detection wavelength = 280 nm, Injection volume = 1 µL.

Figure 2 shows an X-LC chromatogram of the tea sample. Each component is well separated from unknown peaks.

Figure 2. X-LC chromatogram of commercially-available tea beverage
Conditions: the same as in the Figure 1.

Keywords

520015X

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
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