Three-lobed sage (leaf) HPTLC ID - Anisaldehyde reagent, white RT

Three-lobed sage (leaf) (Salvia fruticosa, syn. S. triloba)

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

1. 3µL Sage oil
2. 3µL Clary sage oil
3. 10µL Cineole, thujon (with increasing Rf)
4. 5µL Sage leaf
5. 7µL Sage leaf
6. 10µL Sage leaf
7. 5µL Clary sage leaf
8. 7µL Clary sage leaf
9. 10µL Clary sage leaf
10. 5µL Spanish sage leaf
11. 7µL Spanish sage leaf
12. 10µL Spanish sage leaf
13. 5µL Three-lobed sage leaf
14. 7µL Three-lobed sage leaf
15. 10µL Three-lobed sage leaf 

Reference Sample(s) Reference: Dissolve 5 mg of thujon in 1 mL of methanol. Dissolve 25 µL of cineole in 10 mL of toluene. 

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

Mobile Phase Dichloromethane 

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

Derivatization reagent: Anisaldehyde reagent, Preparation: 170 mL of ice-cooled methanol add 20 mL of glacial acetic acid, 10 mL of sulfuric acid and 1 mL of anisaldehyde, Use: spray, 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: Thujon: violet zone at Rf ~ 0.47; Cineole: violet zone at Rf ~ 0.27.

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 violet zone just below the solvent front. A diffuse violet zone is seen at the position of thujon (black arrow). There is a violet zone at Rf ~ 0.30 and slightly below reference cineole there is another violet zone. A brown zone is seen at Rf ~ 0.20 and there is a blue violet zone just below it.

Test for other species: No yellow zone is seen at Rf ~ 0.50 (Sage leaf, green arrow).

Source: HPTLC Association ^[1]