Source: Botanical Voucher Specimen Library, Alkemists Laboratories^[1]

Source: PlantaPhile^[2]

Source: PlantaPhile^[3]

Thick-walled lignified fiber of Eleutherococcus senticosus root viewed at 400x with Acidified Chloral Hydrate Solution.
Source: Elan M. Sudberg, Alkemist Laboratories^[4]

Large rosettes viewed at 400x under polarized light with Acidified Chloral Hydrate Solution.
Source: Elan M. Sudberg, Alkemist Laboratories^[5]

Eleuthero HPTLC ID - Developed, UV 254 nm

Eleuthero (root) (Eleutherococcus senticosus)

Lane Assignments Lanes, from left to right (Track, Volume, Sample): (2 μL of references, 10 μL of test solutions)

1. 2 μL Eleutheroside E
2. 2 μL Eleutheroside B
3. 10 μL Eleuthero extract 1
4. 10 μL Eleuthero extract 2
5. 10 μL Eleuthero root
6. 5 μL Periploca sepium root 

Reference Sample(s) Reference: Dissolve 1 mg of eleutheroside B and 1 mg of eleutheroside E individually in 1.5 mL of ethanol-water (1:1). 

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

Mobile Phase Chloroform, methanol, water 70:30:4 (v/v/v) 

Sample Preparation Method Sample: Mix 1 g of powdered sample with 5 mL of ethanol-water (1:1) and sonicate for 10 minutes, then centrifuge or filter the solutions and use the supernatants / filtrates as test solutions

Sulfuric Acid Reagent: Preparation: 20 mL of sulfuric acid are mixed with 180 mL of ice-cooled methanol 

Detection Method UV/Vis, Sulfuric Acid Reagent Use: Dip (time 0, speed 5), heat at 100°C for 5 min 

Other Notes System suitability test: After derivatization the fingerprint of the test solution shows a zone corresponding to eleutheroside B and another zone directly below. These two zones appear as two distinct bands, compare lane 2 with lanes 3, 4 and lane 1 with corresponding feature in 3, 4. An arrow marking the distinct bands appears in the illustration in further methodology, below.

Identification: Compare result under UV 254 nm and white RT with reference images in Image Comparison Viewer. The fingerprint of the test solution is similar to that of the corresponding botanical reference sample. Additional weak zones may be present.

Eleuthero: The chromatogram of the test solution shows bands corresponding to eleutherosides E (Rf 0.34) and B (Rf 0.45), and two very strong bands near the application zone. Additional brownish-purple zones are seen between eleutheroside E and the solvent front.

Periploca sepium Adulteration: No band corresponding to eleutheroside B is seen, but strong brown bands at Rf=0.53 and Rf=0.28 are detected.

Further methodology available here, in Identification method of Eleutherococcus senticosus by HPTLC.

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  Eleuthero HPTLC ID - Developed, UV 254 nm

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  Eleuthero HPTLC ID - Sulfuric Acid Reagent, white RT

Source: AHPA Practical, CAMAG HPTLC ^[6]