Guggul Tablets are prepared from Native Guggul Extract or Purified Guggul Extract. Tablets contain NLT 90.0% and NMT 110.0% of the labeled amount of Extract, calculated as the sum of guggulsterones E and Z.
• A. Thin-Layer Chromatographic Identification Test
Standard solution: 10 mg/mL of USP Purified Guggul Extract RS, with heating, in acetonitrile
Sample solution: Powder and transfer a portion of the Tablets equivalent to 100 mg of Extract to a conical flask. Extract three times, each with 25 mL of acetonitrile, in a 55 water bath for 15 min, stirring with a magnetic stirrer, and filter. Evaporate the combined extracts to dryness in a vacuum between 45 and 50, dissolve the residue in 10 mL of acetonitrile, centrifuge, and use the clear supernatant.
Developing solvent system: A mixture of hexane and ethyl acetate (6:4)
Adsorbent: 0.25-mm layer of chromatographic silica gel mixture, typically 20 cm in length
Application volume: 10 µL
Samples: Standard solution and Sample solution
Apply the Samples as bands to a suitable plate. Use a saturated chamber. Develop until the solvent front has moved about about three-fourths the length of the plate, dry the plate, and examine under UV light at 254 nm.
Acceptance criteria: The chromatogram of the Sample solution exhibits bands at RF values of about 0.38 and 0.47, due to guggulsterone E and Z, respectively. Both bands correspond in RF to bands in the chromatogram from the Standard solution.
• B. HPLC Identification Test
Analysis: Proceed as directed in the test for Content of Guggulsterones E and Z.
Acceptance criteria: The chromatogram of the Sample solution exhibits peaks for guggulsterones E and Z at retention times that correspond to those of Standard solution A.
• Content of Guggulsterones E and Z
Mobile phase: A mixture of acetonitrile and water (45:55)
Standard solution A: 10 mg/mL of USP Purified Guggul Extract RS, with heating, in acetonitrile. Pass the solution through a filter of 0.45-µm pore size before injection.
Standard solution B: 0.1 mg/mL of USP Guggulsterone Z RS in acetonitrile. Pass the solution through a filter of 0.45-µm pore size before injection.
Sample solution: Weigh and finely powder NLT 20 Tablets. Transfer a weighed amount of the powder, equivalent to about 10 mg of guggulsterones E and Z, to a conical flask, and extract five times, each with a 20-mL portion of acetonitrile, shake for 1 min, and reflux in a water bath for 30 min, stirring with a magnetic stirrer. Evaporate the combined extracts to dryness in a vacuum at 4550. Dissolve the residue in 50.0 mL of acetonitrile, and pass through a filter of 0.45-µm pore size before injection.
Detector: UV 242 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Flow rate: 2.0 mL/min
Injection size: 20 µL
Column temperature: 27 ± 1
Samples: Standard solution A and Standard solution B [NoteThe relative retention times for guggulsterones E and Z are about 0.69 and 1.0, respectively. ]
Chromatogram similarity: The chromatogram from Standard solution A is similar to the reference chromatogram provided with the lot of USP Purified Guggul Extract RS being used.
Resolution: NLT 2.0 between the guggulsterone Z peak and the peak before, Standard solution A
Tailing factor: NMT 1.5 for the guggulsterone Z peak, Standard solution B
Relative standard deviation: NMT 2.0% for the guggulsterone Z peak (replicate injections), Standard solution B
Samples: Standard solution A, Standard solution B, and Sample solution
Allow Standard solution A to elute for NLT two times the retention time of guggulsterone Z, as determined in Standard solution B. Using the chromatogram of Standard solution A and the reference chromatogram provided with the lot of USP Purified Guggul Extract RS being used, identify the retention times of the peaks corresponding to guggulsterone E and guggulsterone Z.
Calculate the content of guggulsterones E and Z as guggulsterone Z in the portion of Tablets taken:
CI = (rU/rS) × CS × V
Result = CI × (AWT/W) × (100/LE) × (100/L)
Acceptance criteria: 90.0%110.0% of the labeled amount of Extract, calculated as the sum of guggulsterones E and Z
• Disintegration and Dissolution 2040: It meets the requirement for Disintegration only; 30 min, the use of the disk being omitted.
• Weight Variation 2091: Meet the requirements
• Microbial Enumeration Tests 2021: The total aerobic microbial count does not exceed 104 cfu/g, and the total combined yeasts and molds count does not exceed 103 cfu/g.
• Absence of Specified Microorganisms 2022: It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
• Residual Solvents 467: Meets the requirements
• Packaging and Storage: Preserve in well-closed containers, protected from light and moisture, and store at room temperature.
• Labeling: The label states the Latin binominal and, following the official name, the article from which the Tablets were prepared. The label also indicates the amount of Extract, in mg/Tablet, and the content, in mg, of guggulsterones E and Z per 100 mg of Extract.
• USP Reference Standards 11
USP Guggulsterone Z RS
USP Purified Guggul Extract RS
Auxiliary Information Please check for your question in the FAQs before contacting USP.
USP35NF30 Page 1353Pharmacopeial Forum: Volume No. 34(4) Page 1004