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HPLC Analysis of Capsaicin and Dihydrocapsaicin in Capsicum

By Masao Bounashita

Introduction

capsaicin analysis — LC-4000 Series HPLC

The pungent components (which are responsible for the ‘hotness’) contained in the fruit of the genus Capsicum are collectively referred to as the Capsaicinoids. Capsaicin and dihydrocapsaicin in Capsicums constitute 80 – 90% of the pungent components; these compounds are now widely used in medical products and health foods.

It is known that the capsaicinoids are present in differing amounts in the various parts of the capsicum fruit. In this experiment the capsaicinoid content of the pericarp, seed and placenta from each of four types of capsicum was measured – Capsicum frutescens, Serrano, Finger hot and Habanero.

Experimental

Equipment

Pump:	PU-4180-LPG
Column oven:	CO-4061
Autosampler:	AS-4050
Detector:	MD-4010

Conditions

Column:	CrestPak C18S (4.6 mmID x 150 mmL, 5 mm)
Eluent A:	1% Acetic acid/Acetonitrile (50/50)
Eluent B:	Acetonitrile
Gradient condition:	(A/B), 0 min (100/0), 15 min (100/0), 15.05 min (0/100)
	20.00 min (0/100), 20.05 min (100/0) 1 cycle; 35.5 min
Flow rate:	1.0 mL/min
Column temp.:	40°C
Wavelength:	280 nm
Injection volume:	20 mL
Standard sample:	Capsaicin 10, 50, 100 µg/mL in methanol
	Dihydrocapsaicin 10, 50, 100 µg/mL in methanol

Keywords

510003H, capsaicin, Dihydrocapsaicin, capsicum, pepper, Scoville Scale

Results

The chromatograms of 10 µg/mL each of Capsaicin and Dihydrocapsaicin standards are shown in Fig. 1. Good separation was obtained within 12 minutes. The sampling procedure of the actual samples is shown in Fig. 2.

Fig. 1 Chromatogram of Capsaicin and Dihydrocapsaicin standard
1: Capsaicin, 2: Dihydrocapsaicin 2µg each

The chromatograms for each of 4 types of capsicum are shown in Fig. 3. In Table 1 the Capsaicin and Dihydrocapsaicin content in each Capsicum sample (wet weight) are shown. It is known that the contents are very different depending on each part and for the all capsicum, Placenta part contains Capsaicin and Dihydrocapsaicin the most.

Fig. 3 Chromatograms of each of the four species of Capsicum measured. Table 1 shows the Capsaicin and Dihydrocapsaicin content for each sample (wet weight).

Table 1 Capsaicin and Dihydrocapsaicin content in each part of the Capsicum

Sample	Components in each sample [µg/g]
	Capsaicin	Dihydrocapsaicin
Capsicum frutescens
Pericarp	154	120
Seed	713	542
Placenta	9540	7410
Serrano
Pericarp	7.64	4.47
Seed	387	229
Placenta	1080	644
Finger hot
Pericarp	N.D.	N.D.
Seed	2.89	2.87
Placenta	217	160
Habanero
Pericarp	134	43.2
Seed	297	92.5
Placenta	8720	2750
N.D.: Not detected

Conclusion

It is well known that the amount of capsaicinoids present is very different depending on the part of the capsicum fruit and is also widely different between species; as seen here the placenta contains the majority of capsaicin and dihydrocapsaicin, followed by the seed and then the pericarp which may contain little or no capsaicinoids. This simple application demonstrates a reliable sample preparation combined with a routine chromatographic separation to determine the concentration of capsaicinoids.

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.

About the Author

Masao Bounashita

Introduction