Boswellia serrata – Oleo-gum resin

1. Scope
This method identifies dried oleo-gum resin of stems and branches of Boswellia serrata Roxb.
Ex Colebr. by HPTLC fingerprint and discriminates the herbal drugs described under results.
2. Source of method
HPTLC Association
Adopted in USP Dietary Supplements Compendium 2019 as non-USP method.
3. Procedure
Test solution Mix 0.20 g of powdered sample with 4.0 mL of ethanol 96% and
sonicate for 10 minutes, then centrifuge the mixture. Use the
supernatant as test solution.
Reference solutions Standard Solution A: 1 mg/mL of thymol and 4 µL/mL of anethole
in ethanol 96%.
Standard Solution B: 30 mg/mL of USP Powdered Boswellia
serrata Extract RS in ethanol 96%, sonicate for 10 minutes, then
centrifuge the mixture. Use the supernatant as reference solution.
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 2.0 µL of reference solutions and of test solution.
Developing solvent Cyclohexane, diisopropyl ether, glacial acetic acid 6:4:1 (v/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 and carefully mix 170 mL of ice-cooled
methanol with 20 mL of acetic acid and 10 mL of sulfuric acid.
Allow the mixture to cool to room temperature, then add 1 mL of
anisaldehyde (p-methoxy benzaldehyde).
Use: Derivatize (Derivatizer: 3 mL, blue nozzle, spraying level 3),
heat at 100°C for 3 min.
Detection A) Underivatized, UV 254 nm
B) Underivatized, UV 366 nm
C) Underivatized, white light RT
D) Derivatized, UV 366 nm
E) Derivatized, white light RT

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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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 (under white light after derivatization)

 thymol: orange zone at RF ~ 0.59

 anethole: violet zone at RF ~ 0.69

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

Track Sample Origin

1 SST Thymol, USP Anethole RS (with increasing RF) -
2 USP Powdered Boswellia serrata Extract Unknown
3-5 Boswellia serrata – powdered oleo-gum resin Unknown
6-8 Boswellia sacra – powdered oleo-gum resin (same preparation as test solution) Unknown
9-10 Dammar – powdered gum resin (same preparation as test solution) Unknown
11-12 Commiphora mukul – powdered oleo-gum resin (same preparation as test solution) Unknown
13 Commiphora myrrha – powdered oleo-gum resin (same preparation as test solution) Unknown
14 Styrax tonkinensis – powdered oleo-gum resin (same preparation as test solution) Unknown

Version Revision history Released by

1 Created by: VW/26 Nov 2018 MHS/22 Jan 2019
2 Method description changed; Samples labels were harmonized with other M Sharaf/25 Mar 2020
methods: DF/05 Aug 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
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