Source: Tropicos.org. Missouri Botanical Garden. 05 Aug 2013^[1]

Source: Botanical Voucher Specimen Library, Alkemists Laboratories^[2]

Source: Royal Botanic Gardens, Kew.^[3]

Yarrow (flower) HPTLC ID - Anisaldehyde reagent, white RT

Yarrow (flower) (Achillea millefolium)

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

1. 8 µL Yarrow flower 1
2. 16 µL Yarrow flower 1
3. 32 µL Yarrow flower 1
4. 1 µL Yarrow flower 1 (Ph. Eur extr.)
5. 3 µL Yarrow flower 1 (Ph. Eur extr.)
6. 6 µL Yarrow flower 1 (Ph. Eur extr.)
7. 2 µL Guaiazulene
8. 5 µL Cineole
9. 8 µL Yarrow flower 2
10. 16 µL Yarrow flower 2
11. 32 µL Yarrow flower 2
12. 8 µL Yarrow flower 3
13. 16 µL Yarrow flower 3
14. 32 µL Yarrow flower 3 

Reference Sample(s) Reference: Dissolve 25 mg of cineole in 20 mL of tolulene. Dissolve 10 mg of guaiazulene in 20 mL of tolulene. 

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

Mobile Phase Tolulene, ethyl acetate 95:5 (v/v) 

Sample Preparation Method Sample: Mix 1.0 g of powdered sample with 10 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 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 4 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 a bases for setting specifications for quality control purposes.

System suitability test: Cineole: violet zone at Rf ~ 0.38. Guaiazulene: brown zone at Rf ~ 0.88.

Application: 2 µL of references, 16 µL of test solutions

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 at Rf ~ 0.94 right above the zone due to guaiazulene reference substance. Below this zone there is a violet zone at Rf ~ 0.70. There is a reddish-violet zone at Rf ~ 0.43 right above the zone due to cineole reference substance. Below this zone there is a violet zone at Rf ~ 0.29 (black arrows).

Source: HPTLC Association ^[7]