Senna Leaf
(sen' a).
Senna Leaf consists of the dried leaflet of Senna alexandrina Mill. also known as Cassia acutifolia Delile (Alexandrian senna) or C. angustifolia Vahl (Tinnevelly senna) (Fam. Fabaceae). Senna Leaf contains NLT 2.5% of anthraquinone glucosides, calculated as sennosides, on the dried basis.
•  A.
Potassium hydroxide solution:  100 mg/mL of potassium hydroxide in alcohol
Sample:  500 mg of finely powdered Senna Leaf
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.
•  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 Leaf. 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
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 Sample solution, with a suitable spectrophotometer fitted with matched quartz cells, using the Blank.
Calculate the percentage of sennosides in the portion of Senna Leaf taken:
Result = (AU/AS) × CS × (V/W) × 100
AU== absorbance from the Sample solution
AS== absorbance from the Standard solution
CS== concentration of USP Sennosides RS in the Standard solution (mg/mL)
V== volume of water to which powdered Senna Leaf was added, 30 mL
W== weight of powdered Senna Leaf (mg)
Acceptance criteria:  NLT 2.5% of anthraquinone glucosides, calculated as sennosides, on the dried basis
•  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.
•  Microbiological Procedures for Absence of Specified Microorganisms 2022: It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
•  Botanic Characteristics
Unground Alexandrian senna leaf:  Inequilaterally lanceolate or lance-ovate leaflets, frequently broken; from 1.5 to 3.5 cm in length and from 5 to 10 mm in width, unequal at the base, with very short, stout petiolules. The leaflets are acutely cuspidate, entire, brittle, and subcoriaceous, with short and somewhat appressed hairs, few on the upper surface, more numerous on the lower surface, where they occur spreading on the midrib, especially on its lower part. The color is weak yellow to light grayish-green to pale olive. The odor is characteristic.
Unground Tinnevelly senna leaf:  Usually unbroken leaflets, from 2 to 5 cm in length and from 6 to 15 mm in width; acute at the apex; and slightly hairy. The color of the leaves is weak yellow to pale olive.
Powdered senna leaf:  Dusky greenish-yellow to light olive-brown.
Transverse section:  Senna Leaf shows polygonal epidermal cells with straight walls and frequently containing mucilage; numerous, broadly elliptical stomata mostly from 20 to 35 µm in length, usually bordered by two neighbor-cells with their long axes parallel to that of the stoma, and rarely, though more frequently in Alexandrian Senna Leaf, a third epidermal cell at the end of the stoma. The hairs are nonglandular, one-celled, conical, often curved, with thick papillose walls, from 100 to 350 µm in length. Palisade cells in a single layer underlie both surfaces except in the midrib region where they occur only beneath the upper epidermis. A meristele occurs in the midrib composed of several radially arranged fibrovascular bundles, the latter separated by narrow vascular rays and supported above and below by arcs of lignified pericyclic fibers. Calcium oxalate occurs in rosette aggregates in the spongy parenchyma and in six- to eight-sided prisms in the crystal fibers, which lie on the outer surface of each group of pericyclic fibers.
Powdered senna leaf:  Fragments of veins bearing lignified vessels, tracheids, and crystal fibers, isolated hairs, masses of palisade and spongy parenchyma, fragments of epidermis with stomata, free calcium oxalate rosette aggregates, and prisms from 10 to 20 µm in length. In powdered Alexandrian Senna Leaf, the hairs are more numerous than in powdered Tinnevelly Senna Leaf.
•  Articles of Botanical Origin, Foreign Organic Matter 561: The amount of senna stems is NMT 8.0%, and the amount of senna pods or other foreign organic matter is NMT 2.0%.
•  Loss on Drying 731: Dry 1.0 g of finely powdered Senna Leaf at 105 for 2 h: it loses NMT 12.0% of its weight.
•  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
USP Sennosides RS Click to View Structure
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