Capsicum annuum var. annuum L. HPTLC ID - Vanillin Sulfuric Acid White RT

Cayenne (fruit) (Capsicum annuum var. annuum L.)

Lane Assignments Lanes, from left to right (Track, Volume, Sample):

1. 2 μL Capsaicin ~0.1% in Methanol
2. 2 μL Capsicum annuum var. annuum L.-1 (fruit)
3. 2 μL Capsicum annuum var. annuum L.-2 (fruit)
4. 2 μL Capsicum annuum var. annuum L.-3 (fruit)
5. 2 μL Capsicum annuum var. annuum L.-3 (fruit)
6. 2 μL Capsicum annuum var. annuum L.-4 (fruit)
7. 2 μL Capsicum annuum var. annuum L.-5 (fruit)
8. 2 μL Capsaicin ~0.1% in Methanol

Reference materials used here have been authenticated by macroscopic, microscopic &/or TLC studies according to the reference source cited below held at Alkemists Laboratories, Costa Mesa, CA. 

Stationary Phase Silica gel 60, F254, 10 x 10 cm HPTLC plates 

Mobile Phase toluene: ethyl acetate [7/3] 

Sample Preparation Method 0.5g+5ml dichloromethane, sncte/cntrfge/dcnt, evap dry (N2), qs 1.0 ml Toluene 

Detection Method Vanillin/H2SO4 Reagent -> 110° C 5 min -> Visible light 

Reference see British Herbal Pharmacopoeia, 1996

Source: Elan M. Sudberg, Alkemist Laboratories ^[4]

Capsicum annum / Capsicum frutescens HPTLC ID - Dichloroquinone chlorimide reagent and ammonia vapour, white RT

Cayenne pepper (fruit) (Capsicum annum / Capsicum frutescens)

Lane Assignments Lanes, from left to right (Track, Volume, Sample):

1. 3 µL Capsicum annum (Ph.Eur.)
2. 4 µL Capsicum annum (Ph.Eur.)
3. 5 µL Capsicum annum (Ph.Eur.)
4. 3 µL Capsicum annum
5. 4 µL Capsicum annum
6. 5 µL Capsicum annum
7. 2 µL Capsaicin
8. 2 µL Dihydrocapsaicin
9. 2 µL Capsicum frutescens 1
10. 2 µL Capsicum frutescens 2
11. 8 µL Cayenne pepper 1
12. 10 µL Cayenne pepper 1
13. 6 µL Cayenne pepper 2
14. 8 µL Cayenne pepper 2 

Reference Sample(s) Reference: Dissolve 2 mg of capsaicin in 5 mL of methanol. Dissolve 2 mg of dihydrocapsaicin in 5 mL of methanol. 

Stationary Phase Stationary phase, i.e. Silica gel 60, RP-18, F254 

Mobile Phase Water, methanol 20:80 (v/v) 

Sample Preparation Method Sample: Mix 1 g of powdered sample with 10 mL of methanol and sonicate for 10 minutes, then centrifuge or filter the solutions and use the supernatants / filtrates as test solutions.

Derivatization reagent: Dichloroquinone chlorimide reagent Preparation: 50 mg dichloroquinone chlorimide in 200 mL of ethyl acetate Use: Dip (time 0, speed 5), dry with cold air for 5 min, then expose to ammonia vapor 

Detection Method Saturated chamber; developing distance 70 mm from lower edge; relative humidity 33% 

Other Notes Images presented in this entry are examples and are not intended to be used as basis for setting specifications for quality control purposes.

System suitability test: Capsaicin: blue zone at Rf ~ 0.47; Dihydrocapsaicin: blue zone at Rf ~ 0.40

Identification: Compare result with reference images. The fingerprint of the test solution is similar to that of the corresponding botanical reference sample. Additional weak zones may be present. The chromatogram of the test solution shows two blue zones at Rf ~ 0.40 and Rf ~ 0.47.

Source: HPTLC Association ^[5]