Curcuma zanthorrhiza – root and rhizome

1. Scope
This method identifies dried roots and rhizomes of Curcuma zanthorrhiza Roxb. by HPTLC
fingerprint and discriminates related herbal drugs.
2. Authors and adoptions
HPTLC Association.
Adopted in the International HPTLC Atlas for Identification of Herbal Drugs.
3. Procedure
Test solution To 200 mg of powdered rhizome, add 3 mL of methanol, sonicate
for 10 min, centrifuge and use the supernatant as test solution.
Reference solutions 1.0 mg/mL each of bisdesmethoxycurcumin, desmethoxycurcumin,
and curcumin in methanol.
Stationary phase HPTLC Si 60 F254 (Merck)
Application 15 tracks, band length 8 mm, track distance 11.4 mm, distance from
left edge 20 mm, distance from lower edge 8 mm, application
volume 2 µL of reference solutions and test solutions
Developing solvent Toluene, acetic acid 4:1 (v/v)
Developing distance 70 mm from lower edge
Saturation time 20 min, with a saturation pad
Relative humidity 33%, saturated MgCl2
Temperature 22 ± 5°C
Derivatization reagent Anisaldehyde reagent
Preparation: Slowly mix 85 mL of ice-cooled methanol with 10 mL
of acetic acid and 5 mL of sulfuric acid. Allow the mixture to cool to
room temperature, then add 0.5 mL of anisaldehyde (p-methoxy
benzaldehyde).
Use: Derivatize (Derivatizer: 3 mL, blue nozzle, spraying level 3),
heat plate at 100°C for 3 min, let cool down
Detection A) Underivatized, UV 254 nm
B) Underivatized, UV 366 nm
C) Underivatized, white light
D) Derivatized, UV 366 nm
E) Derivatized, white light

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
Page 1 of 3
4. Results
Note: These chromatographic fingerprints are representative of the samples used in this
analysis. Fingerprints obtained may vary from sample to sample. Analysts must validate the
most appropriate fingerprint for their identity standard.
System suitability test (SST)

 Under development
Specific markers used as SST (under UV 366 nm, after derivatization)

 Bisdesmethoxycurcumin: an orange zone at RF ~ 0.19

 Desmethoxycurcumin: an orange zone at RF ~ 0.29
 Curcumin: a red zone at RF ~ 0.42

Origin(s):

Figure 1: HPTLC profiles under UV 254 nm (A), UV 366 nm (B) and white light (C) prior to derivatization, and under
UV 366 nm after derivatization (D), and white light after derivatization (E).

Track Sample Origin

Curcuminoids (bisdesmethoxycurcumin (1 mg/mL), desmethoxycurcumin
1 --
(1 mg/mL), curcumin (1 mg/mL) with increasing RF)
2-4 Curcuma zanthorrhiza rhizome Indonesia (ID)

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
Page 2 of 3
Related herbal drugs

Figure 2: HPTLC profiles under UV 254 nm (A), UV 366 nm (B) and white light (C) prior to derivatization, and under
UV 366 nm after derivatization (D), and white light after derivatization (E).

Track Sample
Curcuminoids (bisdesmethoxycurcumin (1 mg/mL), desmethoxycurcumin (1 mg/mL),
1
curcumin (1 mg/mL) with increasing RF)
2-3 Curcuma zanthorrhiza rhizome
4-5 Curcuma longa rhizome (same test solution preparation)
6-7 Curcuma aromatica rhizome (same test solution preparation)
8-9 Curcuma amada rhizome (same test solution preparation)
10-11 Curcuma comosa rhizome (same test solution preparation)
12-13 Curcuma zedoaria rhizome (same test solution preparation)
14-19 Curcuma spp. unresolved drugs (same test solution preparation)

Version Revision history Released by

1 Created by: TD/06 May 2019 MHS, 09 May 2019

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
Page 3 of 3