Analysis of Capsaicin and Dihydrocapsaicin in Capsicum

February 16, 2021


The pungent components contained in the Capsicum are collectively referred to as the Capsaicinoid. Especially, the Capsaicin and Dihydrocapsaicin in Capsicum constitute 80 – 90% of the pungent components, and are now used in the medical products and healthy food. In addition, it is known that the Capsaicinoid content depends on the part of the Capsicum.

In this article, the Capsaicinoid content of Pericarp, Seed and Placenta each of the 4 kinds of the Capsicum such as the Capsicum frutescens, Finger hot and Habanero was measured and analyzed.

LC-4000 HPLC system


Chromatographic 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.:   40ºC
Wavelength:   280 nm
Injection volume:  20 mL
Standard sample:   Capsaicin (10, 50, 100 mg/mL in Methanol), Dihydrocapsaicin (10, 50, 100 µg/mL in Methanol)


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

Figure 1. Chromatogram of Capsaicin and Dihydrocapsaicin standard sample (1: Capsaicin, 2: Dihydrocapsaicin 2 µg each)

Figure 2. Preparation of the sample

The chromatograms for each 4 kinds of Capsicum are shown in Figure 3. In Table 1, the Capsaicin and Dihydrocapsaicin content in the each Capsicum sample (wet weight) are shown. As shown in Table 1, the contents depends on each part, and Placenta part in all capsicum contains the most Capsaicin and Dihydrocapsaicin comparing to the other parts.

Figure 3. Chromatograms for each part (Pericarp, Seed and Placenta) of capsicum (1: Capsaicin, 2: Dihydrocapsaicin)

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

Sample Contents 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

About the Author

Chromatography Group