Source: MOBOT, Tropicos.org^[1]

Rosemary oil (aerial parts) HPTLC ID - Anisaldehyde reagent, white RT

Rosemary oil (aerial parts) (Rosmarinus officinalis)

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

1. 1 µL Rosemary oil
2. 3 µL Rosemary oil
3. 5 µL Rosemary oil
4. 2 µL Borneol
5. 2 µL Bornyl acetate
6. 5 µL Cineole
7. 3 µL Rosemary oil CT Campher Ph.Eur. 5.0
8. 3 µL Rosemary oil CT Cineole
9. 1 µL Rosemary oil Ph.Eur 4.0
10. 5 µL Rosemary leaf #1
11. 5 µL Rosemary leaf #2
12. 5 µL Rosemary leaf #3
13. 5 µL Rosemary leaf #4
14. 5 µL Rosemary leaf from Peru
15. 5 µL Rosemary leaf from Croatia 

Reference Sample(s) Reference: Individually dissolve 1 mg of borneol and 1 mg of bornyl acetate in 1 mL of methanol. Optional: Dissolve 10 µL of cineole in 1 mL of methanol. 

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

Mobile Phase Ethyl acetate, toluene 5:95 (v/v) 

Sample Preparation Method Sample: Dissolve 20 µL of essential oil in 1 mL of toluene. Optional: mix 0.5 g of powdered dry Rosemary leaf with 5 mL of toluene and sonicate for 10 min, then centrifuge or filter the solutions and use the supernatants/filtrates as test solution.

Derivatization reagent: Anisaldehyde reagent, Preparation: 170 mL of ice-cooled methanol are mixed with 20 mL of acetic acid, 10 mL of sulfuric acid and 1 mL of anisaldehyde, Use: Dip (time 0, speed 5), heat at 100°C for 5 min. 

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: Borneol: brownish zone at ~ 0.21; Bornyl acetate: brown zone at ~ 0.54.

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 purple zone in the upper third. At the position of reference bornyl acetate there is a yellowish brown zone. Below this zone a purple zone is detected. At the position of reference cineole there is a greyish violet zone. At the position of reference borneol a yellowish brown zone is seen, just above and below this zone there is each a greyish violet zone.

Source: HPTLC Association ^[6]