Source: MOBOT, Tropicos.org^[1]

Source: PlantaPhile^[2]

Source: PlantaPhile^[3]

Cowslip Primrose (flower) HPTLC ID - NP reagent and PEG reagent, UV 366 nm

Cowslip Primrose (flower) (Primula veris)

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

1. 2 µL Rutin
2. 2 µL Hyperoside
3. 2 µL Primula veris flower
4. 3 µL Rutin
5. 3 µL Hyperoside
6. 3 µL Primula veris flower 

Reference Sample(s) Reference: Dissolve 2 mg of rutin and 2 mg of hyperoside individually in 10 mL of methanol. 

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

Mobile Phase Ethyl acetate, water, anhydrous formic acid, acetic acid (100:27:11:11 v/v/v/v) 

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

Derivatization reagent: 1.) NP reagent, Preparation: 1 g of diphenylboric acid 2-aminoethyl ester is dissolved in 200 mL of ethyl acetate; 2.) PEG reagent, Preparation: 10 g of polyethylene glycol (Macrogol) 400 is dissolved in 200 mL of dichloromethane, Use: Heat plate at 100°C for 5 min, then dip (time 0, speed 5) in NP reagent, dry in a stream of cold air, dip (time 0, speed 5) in PEG reagent and dry in a stream of cold air. 

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 zone at Rf ~ 0.29 Hyperoside: orange zone at Rf ~ 0.49

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 a green fluorescent zone just above reference substance rutin. At the position of rutin an orange to yellow fluorescent zone is detected. Below the position of rutin there are two orange to yellow fluorescent zones. Below a weak greenish-blue zone is detected and further below a yellow to orange zone is present.

Source: HPTLC Association ^[4]