Source: MOBOT, Tropicos.org^[1]

Source: MOBOT, Tropicos.org^[2]

Avena sativa - Tropicos.org - Flora of China Illustrations vol. 22, fig. 443, 9-11
Source: MOBOT, Tropicos.org^[3]

Source: PlantaPhile^[4]

Oat (herb) HPTLC ID - NP/PEG reagent, UV 366 nm

Oat (herb) (Avena sativa)

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

1. 4 µL Rutin
2. 4 µL Hyperoside
3. 7 µL Oat herb #1
4. 10 µL Oat herb #1
5. 15 µL Oat herb #1
6. 10 µL Oat herb #2
7. 7 µL Barley grass #1
8. 10 µL Barley grass #1
9. 15 µL Barley grass #1
10. 10 µL Barley grass #2
11. 7 µL Wheat grass #1
12. 10 µL Wheat grass #1
13. 15 µL Wheat grass #1
14. 10 µL Wheat grass #2 

Reference Sample(s) Reference: Dissolve 1 mg of rutin in 1 mL of methanol; Dissolve 1 mg of hyperoside in 1 mL of methanol. 

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

Mobile Phase Formic acid, water, methyl ethyl ketone, ethyl acetate 10:10:30:50 (v/v/v/v) 

Sample Preparation Method Sample: Mix 500 mg of powdered sample with 5mL of methanol and sonicate for 10 minutes, then centrifuge or filter the solutions and use the supernatants / filtrates as test solutions.

Derivatization reagent: 1.) NP reagent, Preparation: 1 g of natural products reagent in 200 mL ethyl acetate; 2.) PEG reagent, Preparation: 10 g of polyethylene glycol 400 in 200 mL dichloromethane, Use: Heat plate 3 min at 100°C, then dip (time 0, speed 5) in NP reagent, dry and dip (time 0, speed 5) in PEG reagent. 

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: Rutin: orange fluorescent zone at Rf ~ 0.32. Hyperoside: orange fluorescent zone at Rf ~ 0.50.

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. Between the orange zones corresponding to references rutin and hyperoside the chromatogram of the test solution shows two faint blue zones at Rf ~ 0.42 and Rf ~ 0.46 (yellow arrow) and below them a faint greenish zone. Below reference rutin there are two green zones at Rf ~ 0.25 and Rf ~ 0.18 (green arrow). A prominent red zone is located close to the solvent front.

Test for other species: No zone is seen between the application position and the green zone at Rf ~ 0.18 (Barley grass, red arrow). No green zone is seen at Rf ~ 0.44 (Wheat grass, blue arrow).

Source: HPTLC Association ^[5]