Source: MOBOT, Tropicos.org^[1]

Source: Royal Botanic Gardens, Kew.^[2]

Source: PlantaPhile^[3]

Source: PlantaPhile^[4]

Lovage (root) HPTLC ID - UV 254 nm

Lovage (root) (Levisticum officinale)

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

1. 4 µL Imperatorin, z-Ligustilide (incr. Rf)
2. 4 µL Osthole
3. 4 µL Angelica root 1 (these samples do not comply with Ph.Eur specifications)
4. 4 µL Angelica root 2
5. 4 µL Angelica root 3
6. 4 µL Chinese Angelica root
7. 4 µL Dahurian Angelica root
8. 4 µL Doubleteeth pubescent Angelica root 1
9. 4 µL Doubleteeth pubescent Angelica root 2
10. 4 µL Lovage root 1 (these samples do not comply with Ph.Eur specifications)
11. 4 µL Lovage root 2
12. 4 µL Lovage root 3
13. 4 µL Chinese lovage root (Ligusticum sinensis)
14. 4 µL Chinese lovage root 1 (Ligusticum jeholense)
15. 4 µL Chinese lovage root 2 (Ligusticum jeholense)
16. 4 µL Chinese lovage root (Ligusticum chuanxiong) 

Reference Sample(s) Reference: Dissolve 1 mg each of osthole and imperatorin in 10 mL of methanol. Optional: Dissolve 1 mg of Z-ligustilide in 10 mL of methanol. 

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

Mobile Phase Toluene, ethyl acetate, glacial acetic acid 90:10:1 (v/v/v) 

Sample Preparation Method Sample: Mix 1.0 g of powdered sample with 4 mL of heptane and sonicate for 5 minutes, then centrifuge and filter the solutions and use the filtrates as test solutions.

Derivatization reagent: Sulfuric acid reagent, Preparation: 20 mL sulfuric acid with 180 mL methanol, Use: Dip for 1 s, 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: Imperatorin: greenish fluorescent zone at Rf ~ 0.31 (UV 366 nm); Osthole: blue fluorescent zone at Rf ~ 0.36 (UV 366 nm).

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. Under UV 254 nm the chromatogram of the test solution shows an intense blue fluorescent zone at Rf ~ 0.57 and a weak quenching zone corresponding to reference osthole (Rf ~ 0.36) (red arrows). Under UV 366 nm the chromatogram of the test solution shows an intense blue white fluorescent zone at Rf ~ 0.57 and a weak whitish fluorescent zone with similar Rf-value as reference imperatorin (yellow arrows). After derivatization under white light there are two prominent purple zones below the solvent front. Below reference imperatorin there are two distinct purple zones and below them is an intense greenish zone.

Test for adulteration: Under UV 254 nm no zone is seen at or directly below the position of imperatorin. Under UV 366 nm there is no zone at the position of osthole. After derivatization under white light there are no zones at the position of imperatorin and osthole and just below the position of imperatorin there is no prominent diffuse purple zone (red arrow) (Angelica root, Chinese Angelica root, Doubleteeth pubescent root, Dahurian Angelica root, Chinese Lovage root).

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  Lovage (root) HPTLC ID - UV 254 nm

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  Lovage (root) HPTLC ID - UV 366 nm

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  Lovage (root) HPTLC ID - Sulfuric acid, WRT

Source: HPTLC Association ^[5]