Powdered Andrographis Extract

Printed Document Overview

  • Date Printed: Mon Nov 09 2020, 10:13:04 AM

  • Official Status: Currently Official on 09-Nov-2020

  • Official Date: Official as of 1-May-2020

  • Document Type: Dietary Supplements

  • Document ID: 1GUID-A77016D8-DCBB-4B79-8587-829967F4C0B84_en-US

  • Source: https://online.uspnf.com/uspnf/document/1GUID-A77016D8-DCBB-4B79-8587-829967F4C0B84_en-US

Powdered Andrographis Extract

Definition

  • **Preparation: **

    • Powdered Andrographis Extract is formed by extracting Andrographis using methanol or alcohol.

    • The plant material to extract ratio is between 15:1 and 10:1.

  • Composition Specifications:

    • Contains Not Less Than (NLT) 90.0% and Not More Than (NMT) 110.0% of the labeled amount of diterpene lactones on a dry basis.

    • Diterpene lactones include:

    • Andrographolide

    • Neoandrographolide

    • 14-deoxy-11,12-didehydroandrographolide

    • Andrograpanin

    • The content of 14-deoxy-11,12-didehydroandrographolide is NMT 15% of the total diterpene lactones.

    • May include suitable added substances as carriers.

Identification

A. Thin-Layer Chromatographic Identification Test 〈201〉

  • Standard Solutions Preparation:

    • Standard Solution 1:

    • Use Standard solution A, prepared per the Content of Diterpene Lactones test.

    • Standard Solution 2:

    • Sonicate approximately 15 mg of diterpene lactones equivalent of USP Powdered Andrographis Extract RS in 25 mL methanol for 10–15 min.

    • Centrifuge and use the supernatant.

    • Sample Solution:

    • Similar preparation as Standard Solution 2 with the test sample.

  • Adsorbent: Chromatographic silica gel with an average particle size of 10–15 µm.

  • Application Volume: 10 µL (5–10 mm bands).

  • Developing Solvent System:

    • Chloroform, acetone, toluene (ratio 2:2:1).

  • Spray Reagent:

    • 1% vanillin in alcohol and 10% sulfuric acid in alcohol (1:1).

  • Analysis Procedure:

    • Use a saturated chamber; develop until the solvent reaches approximately 90% up the plate.

    • Dry, spray with the reagent, and heat for 5–10 min at 100°C; examine under visible light.

  • Acceptance Criteria:

    • Sample solution shows three zones: grayish-blue with R values of about 0.4, 0.6, and 0.8 compared to Standard solution 2

    • The andrographolide zone from Standard Solution 1 appears at an R of about 0.4, consistent with the sample solution.

B. Peak Identification in Content Test

  • Content Test Analysis:

    • The Sample solution in the test spot shows a main peak at the retention time of andrographolide from Standard solution A.

    • Identify other diterpene lactone peaks by comparison to Standard solution B and the reference chromatogram for USP Powdered Andrographis Extract RS.

    • Peaks to be identified include:

    • Neoandrographolide

    • 14-deoxy-11,12-didehydroandrographolide

    • Andrograpanin

Composition

Content of Diterpene Lactones

  • Preparation of Solutions:

    • Solution A:

    • Dissolve 0.14 g of potassium dihydrogen phosphate in 900 mL water, add 0.5 mL phosphoric acid, dilute to 1000 mL, mix, filter, and degas.

    • Solution B: Use filtered and degassed acetonitrile.

  • Standard Solutions for Content Calculation:

    • Standard solution A: Prepare by dissolving a specified quantity of USP Andrographolide RS in methanol to achieve about 1.0 mg/mL.

    • Standard solution B: Prepare by transferring an amount of USP Powdered Andrographis Extract RS (about 25 mg), adding 25 mL methanol, heating, diluting to volume with acetonitrile, and mixing.

  • Sample Solution Preparation: Similar to Standard solution B, replace RS with the test sample.

  • Mobile Phase Gradient Table:

    • Time (min) - Solution A% - Solution B%

    • 0 - 95 - 5

    • 18 - 55 - 45

    • 25 - 20 - 80

    • 28 - 20 - 80

    • 35 - 55 - 45

    • 40 - 95 - 5

    • 45 - 95 - 5

Chromatographic System

  • Fitting to Chromatography Standards:

    • Refer to Chromatography 〈621〉 for system suitability.

  • Detection: UV at 223 nm.

  • Column Specifications: 4.6-mm × 25-cm; 5-µm packing L1.

  • Flow Rate: 1.5 mL/min.

  • Injection Size: 20 µL.

  • System Suitability Samples: Standard solution A and Standard solution B.

  • Suitability Requirements:

    • Chromatogram from Standard solution B should match the reference chromatogram from USP Powdered Andrographis Extract RS.

    • Column efficiency: NLT 5000 theoretical plates for Standard solution A.

    • Tailing Factor: NMT 1.5 for andrographolide peak.

    • Relative standard deviation: NMT 2.0% (from replicate injections of andrographolide peak).

    • Resolution: NLT 5 between neoandrographolide and 14-deoxy-11,12-didehydroandrographolide peaks in Standard solution B.

  • Analysis Samples: Standard solution A, Standard solution B, and Sample solution.

    • Calculate percentages of the diterpene lactones using the formula:
      Result=(rrA)×(CW)×5\text{Result} = \left(\frac{r}{r_{A}}\right) \times \left(\frac{C}{W}\right) \times 5

    • Where:

    • rr = peak response for each diterpene lactone from Sample solution.

    • rAr_{A} = peak response for andrographolide from Standard solution A.

    • CC = concentration of USP Andrographolide RS in standard solution A (mg/mL).

    • WW = weight of Powdered Andrographis Extract taken (g).

    • FF = conversion factor for each analyte:

      • 1.00 for andrographolide

      • 3.90 for neoandrographolide

      • 1.45 for 14-deoxy-11,12-didehydroandrographolide

      • 2.65 for andrograpanin.

  • Acceptance Criteria: 90.0%–110.0% on the dried basis for labeled amount of diterpene lactones as a sum of percentages of the components.

Impurities

Organic Impurities

  • Testing Standards:

    • Adhere to the requirements established under Botanical Extracts 〈565〉, addressing general pharmacopeial requirements and pesticide residues.

Specific Tests

  • Loss on Drying 〈731〉: Dry a sample of 2.0 g at 105°C for 3 hours; loss should not exceed 5.0%.

  • Microbial Enumeration Tests 〈2021〉: Total aerobic microbial count must not exceed 10 cfu/g. Total combined yeasts and molds must not exceed 10 cfu/g.

  • Microbiological Procedures for Absence of Specified Microorganisms 〈2022〉: Requirement tests for absence of Salmonella species and Escherichia coli must be met.

Additional Requirements

  • Packaging and Storage:

    • Must be preserved in well-closed containers, protected from light and moisture, and stored at controlled room temperature.

  • Labeling Entailments:

    • Labels must state the Latin binomial and the part of the plant utilized in the extract, along with additional requirements from Botanical Extracts 〈565〉.

  • **USP Reference Standards: **

    • USP Andrographolide RS

    • USP Powdered Andrographis Extract RS

Auxiliary Information

  • For inquiries, refer to FAQs before contacting USP.

  • Contact Person:

    • Expert Committee: Token-BDSHM2020

    • Scientific Liaison: Anton Bzhelyansky

    • Topic/Question: Powdered Andrographis Extract

  • Chromatographic Database Information:

    • Recently appeared in Pharmacopeial Forum, Volume No. 35(5), Page Information: USP43-NF38 - 4762, USP42-NF37 - 4718, USP41-NF36 - 4433

  • Current Document ID: GUID-A77016D8-DCBB-4B79-8587-829967F4C0B84en-US