Ziziphus jujuba var. spinosa Seed

DEFINITION

The article consists of the sun dried ripe seeds of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chow (Family Rhamnaceae) collected in late autumn to early winter. It contains NLT 0.15% of total flavone C-glycosides calculated as the sum of spinosin and 6'''-feruloylspinosin on the dried basis. It contains NLT 0.1% of total triterpenoid glycosides calculated as the sum of jujuboside A and jujuboside B on the dried basis.

SYNONYMS

Ziziphus sativa var. spinosa (Bunge) C.K. Schneid.

Ziziphus spinosa (Bunge) Hu ex F.H. Chen

Ziziphus vulgaris var. spinosa Bunge

POTENTIAL CONFOUNDING MATERIALS

Zizyphus mauritianae Lam.

Ziziphus jujuba Mill.

Hoveniae dulcis Thunb.

Cercis chinensis

SELECTED COMMON NAMES

Chinese: 酸枣仁

English: Spine date seed, sour Chinese date seed, sour jujube seed, sour date seed

Korean: 산조인 (sanjoin)

Pinyin: Suanzaoren, suan zao ren

CONSTITUENTS OF INTEREST

Saponins: Jujubosides, including jujuboside A and jujuboside B

Flavone C-glycosides: spinosin, 6'''-feruloylspinosin

IDENTIFICATION

• A. Botanic Characteristics

Macroscopic: Oblate or flattened ellipsoidal, 5–9 mm long, 5–7 mm wide, about 3 mm thick The surface is purplish-red or purplish-brown, smooth, lustrous. Some show fissures; some show round protrusions at two sides; one end is dented showing a linear hilum. The testa is fragile; endosperm is white with two pale yellow, oily cotyledons.

Microscopic: Transverse section reveals seed coat composed of an upper epidermis, parenchyma and lower epidermis; upper epidermal cells are radially elongated; endosperm is composed of parenchyma containing cluster crystals of calcium oxalate, aleurone grains, and starch grains; cotyledons are composed of parenchyma containing aleurone grains, starch grains, and oil droplets.

• B. HPTLC for Articles of Botanical Origin <203>

Standard solution A: 1 mg/mL of USP Jujuboside A RS (To Come) in methanol

Standard solution B: 1 g of USP Ziziphus jujuba Mill. var. spinosa Seed Powder RS (To Come) in 5 mL of methanol. Sonicate for 10 min on a water bath at 60°, centrifuge, and use the supernatant.

Sample solution:  1 g of Ziziphus jujuba var. spinosa Seed, finely powdered, in 5 mL of methanol. Sonicate for 10 min on a water bath at 60°, centrifuge, and use the supernatant.

Chromatographic system 

Adsorbent: Chromatographic silica gel with an average particle size 5 µm (HPTLC plates)

Application volume: 8 µL each 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: Dichloromethane, glacial-acetic acid, methanol, and water (64:32:12:8)

Developing distance: 6 cm

Derivatization reagent: Anisaldehyde-sulfuric acid reagent – 85 mL of ice-cooled methanol mixed with 10 mL of glacial-acetic acid, 5 mL of sulfuric acid, and 0.5 mL of p-anisaldehyde.

Analysis 

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

Apply the Samples as bands to a suitable HPTLC plate and dry in air. Develop the chromatograms in a saturated chamber, remove the plate from the chamber, and dry. Treat with Derivatization reagent, and heat for 3 min at 100°. Examine under visible light.

System suitability: Under visible light, the chromatogram of Standard solution B exhibits five color bands in the lower-third section. These include with increasing R_F: three olive green-brownish bands, the one with the highest R_F is the most intense; a violet band at an R_F corresponding to the band due to jujuboside A in Standard solution A; and a faint olive-green band. The Standard solution B exhibits two minor bands in the middle-third section: a pinkish-violet band, corresponding to jujuboside B, and a faint brown band below jujuboside B.  In the upper-third section, the Standard solution B exhibits two distinct violet bands.

Acceptance criteria: Under visible light, the chromatogram of the Sample solution exhibits a violet band corresponding to the band due to jujuboside A in Standard solution A, and the following bands corresponding to similar bands in Standard solution B: three olive green-brownish bands, the one with the highest R_F is the most intense; a faine olive-green band above the jujuboside A; two minor bands in the middle-third section: a pinkish-violet band due to jujuboside B, and a faint brown band below jujuboside B; and two distinct violet bands in the upper-third section.

ASSAY

• Content of Saponins

CALL FOR SUBMISSION OF VALIDATED INFORMATION

Additional information including validation data will be required to complete the development of the Assay. For requirements, please see under "Composition" and related sections of the guidelines document “Monographs in the Herbal Medicines Compendium” at http://hmc.usp.org/about/general-noticesresources

Interested parties are encouraged to submit their proposals to complete the monograph.

CONTAMINANTS

• 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 the absence of Salmonella species and Escherichia coli

SPECIFIC TESTS

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

• Articles of Botanical Origin, Alcohol-Soluble Extractives, Method 1 <561>: NLT 9.0%

• Loss on Drying <731>

Analysis: Dry 1.0 g of Ziziphus jujuba var. spinosa Seed, finely powdered, at 105° for 5 h.

Acceptance criteria: NMT 10%

• Articles of Botanical Origin, Total Ash <561>

Analysis: 2.0 g of Ziziphus jujuba var. spinosa Seed, finely powdered

Acceptance criteria: NMT 7%

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, following the official name, the part of the plant contained in the article.

• USP Reference Standards <11>

USP Jujuboside A RS (To Come)

USP Ziziphus jujuba Mill. var. spinosa Seed Powder RS (To Come)