Senna Pods
DEFINITION
Senna Pods are the dried ripe fruits of Senna alexandrina Mill. also known as Cassia acutifolia Delile (Alexandrian senna) or C. angustifolia Vahl (Tinnevelly senna) (Fam. Fabaceae). Senna Pods contain NLT 3.4% (Alexandrian senna) and NLT 2.2% (Tinnevelly senna) of anthraquinone glucosides, calculated as sennosides, on the dried basis.
IDENTIFICATION
• A.
Potassium hydroxide solution:
100 mg/mL of potassium hydroxide in alcohol
Sample:
500 mg of finely powdered Senna Pods
Analysis:
Add 10 mL of Potassium hydroxide solution to the Sample. Boil for 2 min, dilute with 10 mL of water, and filter. Acidify the filtrate with hydrochloric acid. Shake it with ether, remove the ether layer, and shake it with 5 mL of 6 N ammonium hydroxide.
Acceptance criteria:
The aqueous layer is colored orange or bluish-red.
ASSAY
• Procedure
[Note—Conduct all sample preparations with minimal exposure to subdued light, and use low-actinic glassware to protect solutions from light. ]
Ferric chloride solution:
105 mg/mL of ferric chloride
Methanolic magnesium acetate solution:
5 mg/mL magnesium acetate in methanol
Sodium bicarbonate solution:
5 mg/mL of sodium bicarbonate
Standard solution:
0.13 mg/mL of USP Sennosides RS in Sodium bicarbonate solution
Sample solution:
Weigh and pulverize 10 g of Senna Pods. Transfer 0.15 g to a 100-mL round-bottom flask. Add 30 mL of water. Mix, weigh, attach a condenser, and reflux in a water bath for 15 min. Cool to room temperature, weigh, and adjust to the original weight with water. Centrifuge, and transfer 20.0 mL of the supernatant to a 150-mL separatory funnel. Add 0.1 mL of diluted hydrochloric acid, and shake with three quantities, each of 15 mL, of chloroform. Allow to separate, and discard the chloroform layer after each addition. Add about 0.1 g of sodium bicarbonate, shake for 3 min, and centrifuge. Use the supernatant as the Sample solution.
Instrumental conditions
Mode:
Vis
Wavelength:
515 nm
Cell:
Quartz
Blank:
Methanol
Analysis
Samples:
Standard solution, Sample solution, and Blank
Transfer 10.0 mL each of the Standard solution and the Sample solution to separate 100-mL round-bottom flasks equipped with condensers. Add 20 mL of Ferric chloride solution, and mix. Reflux in a water bath for 20 min. Add 1 mL of hydrochloric acid, and reflux for an additional 20 min, with frequent shaking, to dissolve the precipitates. Cool to room temperature, transfer the mixtures to separate 100-mL separatory funnels, and shake with three quantities, each of 25 mL, of ether previously used to rinse the flasks. Combine the ether extracts, mix, and wash with two quantities, each of 15 mL, of water. Transfer the ether layers to separate 100-mL volumetric flasks. Dilute with ether to volume, and mix. Evaporate 10.0 mL of the ether extracts to dryness, and dissolve the residue in 10.0 mL of Methanolic magnesium acetate solution. Determine the absorbance of the resulting solution from the Standard solution and the Sample solution, with a suitable spectrophotometer fitted with matched quartz cells, using the Blank.
Calculate the percentage of sennosides in the portion of Senna Pods taken:
Result = (AU/AS) × CS × (V/W) × 100
| AU | = | = absorbances from the Sample solution |
| AS | = | = absorbances from the Standard solution |
| CS | = | = concentration of USP Sennosides RS in the Standard solution (mg/mL) |
| V | = | = volume of water to prepare the Sample solution, 30 mL |
| W | = | = weight of powdered Senna Pods taken (mg) |
Acceptance criteria:
NLT 3.4% (Alexandrian senna) and NLT 2.2% (Tinnevelly senna), of anthraquinone glucosides, calculated as sennosides, on the dried basis
CONTAMINANTS
• Microbial Enumeration Tests 
2021
:
The total 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.
2021:
The total 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.
• Microbiological Procedures for Absence of Specified Microorganisms 2022:
It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
2022:
It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
SPECIFIC TESTS
• Botanic Characteristics
Macroscopic
Unground Alexandrian senna pods:
Occur as flattened, reniform, membranous, leathery pods; green to greenish-brown with brown patches at the positions corresponding to the seeds; from 40 to 50 mm in length and at least 20 mm wide; at one end is a stylar point and at the other a short stalk. The pods contain six or seven flattened and obovate seeds, green to pale brown, with a continuous network of prominent ridges on the testa.
Unground Tinnevelly senna pods:
Occur as flattened, slightly reniform, membranous, leathery pods; brown to yellowish-brown with brown patches at the positions corresponding to the seeds; slightly longer but narrower than Alexandrian senna pods, from 35 to 60 mm in length and from 14 to 18 mm wide; at one end is a stylar point and at the other a short stalk. The pods contain 5–8 flattened and obovate seeds, green to pale brown, with incomplete, wavy, transverse ridges on the testa.
Powdered senna pods:
Brown powder
Microscopic
Senna pods:
Senna Pods show epicarp with very thick cuticulized polygonal cells, occasional anomocytic or paracytic stomata, and very few nonglandular, one-celled, conical, often curved hairs, with thick papillose walls, from 100 to 350 µm in length; mesocarp consists of parenchymatous tissue containing a layer of calcium oxalate prisms and vascular bundles partially enclosed by fibers; endocarp with two crossed layers of fibers; seeds with a subepidermal layer of palisade cells with thick outer walls and endosperm of polyhedral cells with mucilaginous walls.
Powdered senna pods:
Polygonal cells with occasional small numbers of nonglandular hairs and anomocytic or paracytic stomata, fibers in two crossed layers accompanied by a crystal sheath of calcium oxalate prisms, isolated hairs, masses of palisade cells of the seeds, clusters, and prisms of calcium oxalate.
• Articles of Botanical Origin, Total Ash 561:
NMT 9.0%
561:
NMT 9.0%
• Loss on Drying 731:
Dry 1.0 g of finely powdered Senna Pods at 105
for 2 h: it loses NMT 12.0% of its weight.
731:
Dry 1.0 g of finely powdered Senna Pods at 105 for 2 h: it loses NMT 12.0% of its weight.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve against attack by insects and rodents. Store protected from light and moisture at room temperature.
• Labeling:
The label states the Latin binomial and, following the official name, the part of the plant contained in the article.
• USP Reference Standards 11
11
USP Sennosides RS 
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Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Maged H. Sharaf, Ph.D.
Principal Scientific Liaison 1-301-816-8318 |
(DS2010) Monographs - Dietary Supplements |
| 2021 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
| 2022 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 4626
Pharmacopeial Forum: Volume No. 32(1) Page 140