Sphaeranthus indicus Aerial Parts

Sphaeranthus indicus Aerial Parts

Proposed For Development Version 0.1

Sphaeranthus indicus Aerial Parts

 


 

DEFINITION

 

The article consists of aerial parts of Sphaeranthus indicus L. (Family: Asteraceae), gathered shortly before or during flowering. It contains NLT 0.14% of sphaeranthanolide calculated on the dried basis.

 

SYNONYMS

Sphaeranthus hirtus Willd.

Sphaeranthus mollis Roxb.

Sphaeranthus mollis Roxb. ex DC.

 

POTENTIAL CONFOUNDING MATERIALS

None known

 

SELECTED COMMON NAMES

English: East Indian globe thistle

Hindi: Gorakhmundi, mundi

Malayalam: Adakkamanian, mirangani

 

CONSTITUENTS OF INTEREST

Sesquiterpene glycoside: Sphaeranthanolide

 

IDENTIFICATION

• A. BOTANICAL CHARACTERISTICS

Macroscopic: Inflorescence (capitula) a coupound head, solitary, terminal on branches, 1-1.6 cm diam, globose or globose-ovoid, pink to purple, ebracteate, peduncled; peduncle 1-4.5 cm long, stout, glandular-pubescent, with toothed wings. Partial heads small, sessile, numerous, heterogamous of few florets, aggregated on a large, common receptacle forming head inflorescence (capitulum). Ivolucres of partial heads, 4 mm long, campanulate, margin toothed; bracts persistent, linear, acuminate, subpaleaceous, ciliate on the upper half. Florets all tubular; marginal and central florets with dark purple corolla; female 6-9, 3-toothed, filiform; 2-3 disc florets hermaphrodite, with 5-toothed corolla, the latter much swollen in lower half. Anther bases sagittate.  Style arms shortly bifid or entire. 

Microscopic: The bract in surface view is composed of thin walled, elongated, rectangular to polygonal-shaped parenchyma cells; marginal cells elongated in to hair-like projections. Vascular strand in centre of spiral elements. A few monoclinic calcium oxalate crystals and starch grains observed in cells. The corolla semitransparent of rectangular to polygonal parenchyma cells showing presence of few crystals. The anthers dark yellow in colour; the cells 2-3 times more elongated than width, thin walled, a few showing oval-spherical-shaped starch grains.  Filament cells comparatively smaller, rectangular, and parenchymatous. The pollen grains spherical, spinous. The gynoecium syncarpous of 2 carpels; ovary superior, oval-shaped, unilocular, with basal placentation; style short; stigma sessile.  Ovary wall multi-layered, 4-6 cells in thickness; cells polygonal, thin-walled. Starch grains few; locule pentagonal.

 

B. Thin-Layer Chromatography

Standard solution A: 0.1 mg/mL of USP Sphaeranthanolide RS in methanol

Standard solution B: 10 mg/mL of USP Sphaeranthus indicus Aerial Parts RS in methanol

Sample solution: Sonicate 5 g of Sphaeranthus indicus Aerial Parts, finely powdered, in 50 mL methanol. FIlter and use filtrate.

Chromatographic system

(See Chromatography <621>, Thin-Layer Chromatography.)

Adsorbent: Use a suitable chromatographic material with an average particle size of 5 µm (HPTLC plates).

Application volume: 10 µL of Standard solution A, Standard solution B, and Sample solution; as 8-mm bands

Relative Humidity: Condition the plate to a relative humidity of about 33% using a suitable device

Developing solvent system: Chloroform and methanol (8:2)

Developing distance: 8 cm

Derivatization reagent: A mixture of 9 mL of ice-cooled methanol, 1 mL of sulfuric acid, mixed in this order.

Analysis

Samples: Standard solution A, Standard solution B, and Sample solution

Apply the samples as bands to a suitable high performance thin-layer chromatographic plate and dry in air. Develop the chromatograms in a saturated chamber, remove the plate from the chamber and dry. Treat the plate with Derivatization reagent, heat for 5 min at 100° and examine under visible light.

System suitability: Under visible light, the chromatogram of the Standard solution B exhibits purple or pink band in the middle of the chromatogram in position and color similar to as sphaeranthanolide band in Standard solution A. A broad drak brown band near origin, one light purple band in two thirds of chromatogram (about RF 0.75) and a blue band near front.

Acceptance criteria: Under visible light, the chromatogram of the Sample solution exhibits a pale pink bands similar in position and color to sphaeranthanolide in Standard solution A. A broad light brown band near origin and a blue band near front.

 

D. HPLC

Analysis: Proceed as directed in the Assay for Content of  Sphaeranthanolide

Acceptance criteria: The chromatogram of the Sample solution exhibits peaks at the retention time corresponding to the peak due to Sphaeranthanolide in Standard solution B.

 

ASSAY

• Content of Sphaeranthanolide

Solution A: Acetonitrile

Solution B: Water

Mobile phase: See Table 1.

 

Table 1

Time
(min)

Solution A
(%)

Solution B
(%)

0

5

95

20

100

0

25

100

0

27

5

95

30

5

95

 

Standard solution A: 0.01 mg/mL of USP Sphaeranthanolide RS in methanol

Standard solution B: To come.

Sample solution: Transfer about 2.50 g of Sphaeranthus indicus Aerial Parts, finely powdered and accurately weighed, to a flask, add 20 mL of methanol, and shake by mechanical means for 2 hours at 60 °C. Cool to room temperature, and pass through filter paper into 50 mL volumetric flask. Wash the flask and the residue on the filter with methanol, dilute the volume with the washings, and mix. Further transfer 1 mL of above solution to 10 mL volumetric flask and dilute with methanol to volume to obtain solution having a known concentration of about 5 mg/mL. Pass solution through membrane filter of 0.45 μm pore size, discard the first few mL of the filtrate.

Chromatographic system

(See Chromatography <621>, System Suitability.)

Detector: UV 206 nm

Column: 4.6-mm × 25-cm; L1 (100 Å)

Flow rate: 1.0 mL/min

Injection volume: 10 µL

System suitability

Sample: Standard solution

Suitability requirements

Chromatogram similarity: To come.

Theoretical plates: NLT 2000

Tailing factor: NMT 2.0

Relative standard deviation: NMT 2.0%

Analysis

Samples: Standard solution(s) and Sample solution

Calculate the percentage of sphaeranthanolide in the portion of Sphaeranthus indicus Aerial Parts taken:

 

Result = (rU/rS) × (CS/CU× P

 

rU   = peak area for sphaeranthanolide from the Sample solution

rS   = peak area for sphaeranthanolide from the Standard solution

CS  = concentration of the Standard solution, (mg/mL)

CU  = concentration of the Sample solution (mg/mL)

P   = percent purity of sphaeranthanolide

 

Acceptance criteria: NLT 0.14% on dried basis

 

CONTAMINANTS

• Elemental Impurities—Procedures <233>

Acceptance criteria

Arsenic: NMT 2.0 µg/g

Cadmium: NMT 0.5 µg/g

Lead: NMT 5 µg/g

Mercury: NMT 0.2 µg/g

• Articles of Botanical Origin, General Method for Pesticide Residues Analysis <561>: Meets the requirements

• Microbial Enumeration Tests <61>: The total aerobic bacterial count does not exceed 105 cfu/g, the total combined molds and yeasts count does not exceed 103 cfu/g, and the bile-tolerant Gram-negative bacteria does not exceed 103 cfu/g.

• Tests for Specified Microorganisms <62>: Meets the requirements of the tests for absence of Salmonella species and Escherichia coli

 

SPECIFIC TESTS

 

• Articles of Botanical Origin, Foreign Organic Matter <561>: NMT 2.0%

• Loss on Drying <731>:

Analysis: Dry 1 g Sphaeranthus indicus Aerial Parts, finely powdered, at 105° for 2 h.

Acceptance criteria: NMT 15%

• Articles of Botanical Origin, Total Ash <561>

Acceptance criteria: NMT 20%

• Articles of Botanical Origin, Acid-Insoluble Ash <561>: NMT 10%

 

ADDITIONAL 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 binomial and the part(s) of the plant contained in the article.

• USP Reference Standards <11>

USP Sphaeranthus indicus Aerial Parts Dry Extract RS (To come)

USP Sphaeranthanolide RS (To come)

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Other Versions

Proposed For Comment Version 0.2