C:
Thin-Layer Chromatographic Identification Test 
201
—
Test solution—
Transfer 0.5 g of finely powdered Myrrh to a 10-mL centrifuge tube, add 2 mL of alcohol, shake for 1 minute, centrifuge, and filter. Apply 2 µL of the filtrate to the plate.
Standard solution—
Dissolve accurately weighed quantities of (E)-anethole, linalool, (–)-bornyl acetate, and (R)-(–)-carvone in toluene, and dilute quantitatively, and stepwise if necessary, with toluene to obtain a solution having known concentrations of about 7 µg per mL of (E)-anethole, 8 µg per mL of linalool, and 10 µg per mL each of (–)-bornyl acetate and (R)-(–)-carvone. Apply 1 µL to the plate.
Developing solvent system:
a mixture of toluene and ethyl acetate (93:7).
Spray reagent—
Dissolve 0.5 mL of p-anisaldehyde in 10 mL of glacial acetic acid, add 85 mL of methanol, and mix. Carefully add 5 mL of sulfuric acid, and mix. [NOTE—Prepare fresh immediately before use.]
Procedure—
[NOTE—Wash the plate in
Developing solvent system, and air-dry prior to use.
] Proceed as directed in the chapter. Spray the plate with
Spray reagent, heat in an oven at 100
for 5 minutes, and examine in white light. The chromatogram of the
Standard solution exhibits four well-resolved spots: an olive-brown spot due to (
E)-anethole at an
RF value of about 0.6; an orange-brown spot due to (–)-bornyl acetate at an
RF value of about 0.5; a reddish brown spot due to (
R)-(–)-carvone at an
RF value of about 0.4; and a deep gray spot due to linalool at an
RF value of about 0.2. The chromatogram of the
Test solution exhibits an intense purplish red spot at an
RF value of about 0.7 and two moderately intense purplish red spots at
RF values of about 0.5 and 0.4. The chromatogram of the
Test solution may exhibit other spots of varying intensities, including a spot at the origin.
D:
Thin-Layer Chromatographic Identification Test 201—
Test solution—
Transfer 0.5 g of finely powdered Myrrh to a test tube containing 5.0 mL of alcohol, and warm the mixture in a water bath for 2 to 3 minutes. Cool, and filter.
Standard solution—
Dissolve accurately weighed quantities of (E)-anethole and thymol in alcohol, and dilute quantitatively, and stepwise if necessary, with alcohol to obtain a solution having known concentrations of about 4 µg of (E)-anethole per mL and about 1 mg of thymol per mL.
Developing solvent system:
a mixture of toluene and ethyl acetate (98:2).
Procedure—
Proceed as directed in the chapter. Allow the plate to air-dry, and examine under UV light at 365 nm. The chromatogram of the Test solution shows no blue to violet fluorescent zones in the lower third of the chromatogram (absence of Commiphora mukul).
