Cascara sagrada bark (Rhamnus purshiana)

1. Scope
This method identifies Cascara sagrada bark (Rhamnus purshiana D.C. syn. Frangula
purshiana (DC.) J.G. Cooper) by HPTLC fingerprint.

2. Source of method
Modified from Ph.Eur 7.4 monograph Cascara, Test A for Other species of Rhamnus
(change in sample preparation, additional reference standard, change in preparation of
derivatization reagent)

3. Procedure
Sample preparation: Mix 500 mg of powdered sample (100 mg of
powdered extract) with 10 mL of ethanol – water
(7:3) and sonicate for 10 minutes, then centrifuge or
filter the solutions and use the supernatants /
filtrates as test solutions.

Reference substances: Dissolve 1.0 mg each of aloin (=barbaloin) and aloe

emodin individually in 10 mL of methanol.

Stationary phase: HPTLC Si 60 F254

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

Mobile phase: Ethyl acetate, methanol, water 100:17:13 (v/v/v)

Development: - Saturated chamber

- Developing distance 70 mm from lower edge
- Relative humidity 33%

Derivatization reagent: KOH reagent

Preparation: 20 g of potassium hydroxide are
dissolved in 200 mL of methanol (prepare in an ice-
bath).
Use: Dip plate (time 0, speed 5) in reagent and heat
plate at 105°C for 10 min.

Documentation: 1.) KOH reagent, white RT

2.) KOH reagent, UV 366 nm

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A non-profit organization dedicated to the promotion of HPTLC in plant analysis and other analytical fields • www.hptlc-association.org
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4. Results
Note: The images presented in this section are examples and are not intended to be used as
basis for setting specifications for quality control purposes.

Fig. 1) KOH reagent, white RT Fig. 2) KOH reagent, UV 366 nm

1 2 3 4 5 1 2 3 4 5
Track Volume Sample
1 2 µL Aloin (Barbaloin)
2 2 µL Aloe emodin
3 2 µL Cascara sagrada bark
4 2 µL Cascara sagrada extract 1
5 2 µL Cascara sagrada extract 2

System suitability test (UV 366 nm)

Aloin: a yellow zone at Rf ~ 0.45.
Aloe emodin: a red zone at Rf ~ 0.81.

Identification
Compare result with reference. The fingerprint of the test solution is similar to that of the
corresponding botanical reference sample. Additional weak zones may be present.

White RT (KOH reagent): The chromatogram obtained with the test solution shows several
reddish-brown zones with different intensities: there are 4 faint zones, 3 being situated at
about the mid-point of the chromatogram and 1 in the lower third and there is a strong zone
in the upper third of the chromatogram.

UV 366 (KOH reagent): The chromatogram obtained with the test solution shows in the
central part an intense yellowish-brown fluorescent zone due to aloin. In the upper part there
is a red fluorescent zone similar in color and position to the zone due to emodin in the
chromatogram obtained with the reference solution. In addition, the chromatogram obtained
with the test solution shows, above the zone due to aloin, another yellowish-brown
fluorescent zone. Below the zone due to aloin there is a series of blue and green fluorescent
zones (cascarosides).

International Association for the Advancement of High Performance Thin Layer Chromatography
A non-profit organization dedicated to the promotion of HPTLC in plant analysis and other analytical fields • www.hptlc-association.org
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