Hibiscus sabdariffa HPTLC ID - Thymol Sulfuric Acid Reagent UV 365 nm

Hibiscus (flower) (Hibiscus sabdariffa)

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

1. μL Epicatechin ~0.1% in CH3OH
2. μL Hibiscus sabdariffa-1 (flower)
3. μL Hibiscus sabdariffa-2 (flower)
4. μL Hibiscus sabdariffa-3 (flower)
5. μL Hibiscus sabdariffa-3 (flower)
6. μL Hibiscus sabdariffa-4 (flower)
7. μL Hibiscus sabdariffa-5 (flower)
8. μL Catechin ~0.1% in CH3OH

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

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

Mobile Phase tolulene: ethyl formate: formic acid [5/4/1] 

Sample Preparation Method 0.5 g + 5mL 70%gr EtOH sonicate/heat @ 50° C~1/2 hr, evap w N2, qs 1mL CH3OH 

Detection Method Thymol Sulfuric acid spray -> 115° C 10 min -> UV 365 nm 

Reference see Thin-Layer Chromatography Jork, Funk Wimmer P421-424

Source: Elan M. Sudberg, Alkemist Laboratories ^[2]

Roselle (flower) HPTLC ID - White RT

Roselle (flower) (Hibiscus sabdariffa)

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

1. 1.0 µL Roselle flower 1
2. 3.0 µL Roselle flower 1
3. 6.0 µL Roselle flower 1
4. 0.5 µL Roselle flower 1 (Ph. Eur. extract.)
5. 1.0 µL Roselle flower 1 (Ph. Eur. extract.)
6. 2.0 µL Roselle flower 1 (Ph. Eur. extract.)
7. 2.0 µL Quinaldine red
8. 2.0 µL Patent blue V
9. 8.0 µL Roselle flower 2
10. 16.0 µL Roselle flower 2
11. 3.0 µL Roselle flower 3
12. 6.0 µL Roselle flower 3
13. 3.0 µL Roselle flower 4
14. 6.0 µL Roselle flower 4 

Reference Sample(s) Reference: Dissolve 2.6 mg of patent blue V in 10 mL of ethanol. Dissolve 2.8 mg of quinaldine red in 10 mL of ethanol. 

Stationary Phase Stationary phase, i.e. Silica gel 60, F254 

Mobile Phase 1-Butanol, formic acid, water 40:10:12: (v/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. 

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: Patent blue V: blue zone at Rf ~ 0.36; Quinaldine red: orange zone at Rf ~ 0.63.

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 test solution shows two violet-blue zones at Rf ~ 0.33 and Rf ~ 0.40.

Source: HPTLC Association ^[3]