Cullen corylifolium Fruit Dry Extract

Cullen corylifolium Fruit Dry Extract

Proposed For Development Version 0.1

Cullen corylifolium Fruit Dry Extract

 


 

DEFINITION

 

The article consists of the dried fruit of Cullen corylifolium (L.) Medik., formerly known as Psoralea corylifolia (Family Fabaceae). The fruit may be misidentified as seeds. It contains NLT 90.0% and NMT 110.0% of the labeled amount of psoralen on the dried basis.

 

POTENTIAL CONFOUNDING MATERIALS

Senna tora (L.) Roxb.

 

CONSTITUENTS OF INTEREST

Furocoumarin: Psoralen

 

IDENTIFICATION

• A. HPTLC for Identification of Articles of Botanical Origin <203>

Standard solution A: 1.0 mg/mL of USP Psoralen RS in methanol

Standard solution B: 50 mg/mL of USP Cullen corylifolium Fruit Dry Extract RS in methanol. Sonicate for 10 min, centrifuge or filter, and use the supernatant or the filtrate.

Sample solution: Sonicate about 0.5 g of Cullen corylifolium Fruit Dry Extract in 10 mL of methanol for 10 min, centrifuge or filter, and use the supernatant or filtrate.

Chromatographic system

Adsorbent: Chromatographic silica gel F254 mixture with an average particle size of 5 µm.

Application volume: 5 µL of Standard solution A, 2 µL of Standard solution B, and 5 µL of 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: Toluene, ethyl acetate, and formic acid (80:18:2)

Developing distance: 6 cm

Analysis

Samples: Standard solution AStandard solution B, and Sample solution

  Apply the Samples as bands and dry in air. Develop in a saturated chamber (20 min with filter paper), remove the plate from the chamber, and dry in air. Examine under UV light at 254 and 365 nm.

System suitability: Under UV 254 nm, the chromatogram of Standard solution B exhibits one intense black band and two black bands in the upper half of the chromatogram, with the lower black band near the middle of the chromatogram similar in RF and color to the psoralen band in the chromatogram of Standard solution A. About eight additional black bands seen in the lower half of the chromatogram. One bluish band appears near RF 0.3. Under UV 365 nm, a blue fluorescent band appears in the upper half of the chromatogram similar in RF and color to the psoralen band in the chromatogram of Standard solution A. A black band, faint blue band, and two orange bands, with increasing RF above the psoralen band appear in Standard solution B. A bright blue fluorescent band and a less bright fluorescent band appear below the psoralen band. Below the less fluorescent band, with decreasing RF, a dark intense black band and an intense bright blue fluorescent band appear in Standard solution B.

Acceptance criteria: Under UV 254 nm, the chromatogram of Sample solution exhibits one intense black band and two black bands in the upper half of the chromatogram, with the lower black band near the middle of the chromatogram similar in RF and color to the psoralen band in the chromatogram of Standard solution A. About eight additional black bands seen in the lower half of the chromatogram. One bluish band appears near RF 0.3. Under UV 365 nm, a blue fluorescent band appears in the upper half of the chromatogram similar in RF and color to the psoralen band in the chromatogram of Standard solution A. A black band, faint blue band, and two orange bands, with increasing RF above the psoralen band appears in Sample solution. A bright blue fluorescent band and a less bright fluorescent band appear below the psoralen band. Below the less fluorescent band, with decreasing RF, a dark intense black band, an intense bright blue fluorescent band appears in the Sample solution.

 

• B. HPLC

Analysis: Proceed as directed in the test for Content of Psoralen.

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

 

ASSAY

• Content of Psoralen

Solution A: 0.1% trifluoroacetic acid in water

Solution B: 0.1% trifluoroacetic acid in acetonitrile

Mobile phase: See Table 2.

 

Table 2

Time
(min)

Solution A
(%)

Solution B
(%)

0

90

10

5

85

15

8

70

30

15

40

60

22

40

60

25

10

90

30

10

90

32

90

10

35

90

10

 

Standard solution A: 1 mg/mL of USP Psoralen RS in methanol

Standard solution B: 5 mg/mL of USP Cullen corylifolium Fruit Dry Extract RS in methanol. Sonicate, and pass through a membrane filter of 0.45-µm or finer pore size.

Sample solution: Transfer 0.5 g of Cullen corylifolium Fruit Dry Extract, accurately weighed, to a 250-mL volumetric flask and add 70 mL of methanol. Reflux at 80° for 15 min and transfer the extract into a 100-mL volumetric flask. Repeat the extraction process with another 20 mL of methanol and add the extract to the volumetric flask. Dilute with methanol to volume. Pass through a membrane filter of 0.45-µm pore size.

Chromatographic system

(See Chromatography <621>, System Suitability.)

Detector: UV 290 nm

Column: 4.6-mm × 10-cm; 2.7-µm packing L1

Column temperature: 40°

Flow rate: 1.0 mL/min

Injection volume: 10 µL

System suitability

Samples: Standard solution A and Standard solution B

Suitability requirements

Chromatogram similarity: The chromatogram of Standard solution B is similar to the reference chromatogram provided with the lot of USP Cullen corylifolium Fruit Dry Extract RS being used.

Resolution: NLT 2.0 between the psoralen peak and the peak after, Standard solution B

Tailing factor: NMT 2.0 for the psoralen peak, Standard solution A

Relative standard deviation: NMT 2.0% for the psoralen peak in repeated injections, Standard solution A

Analysis

Samples: Standard solution AStandard solution B, and Sample solution

Using the chromatograms of Standard solution AStandard solution B, and the reference chromatogram provided with the lot of USP Cullen corylifolium Fruit Dry Extract RS being used, identify the retention times of the peaks corresponding to psoralen in the Sample solution.

Calculate the percentages of psoralen in the portion of Cullen corylifolium Fruit Dry Extract taken:

 

Result = (rU/rS) × (CS/CU) × 100

 

rU   = peak area of psoralen from the Sample solution

rS   = peak area of psoralen from Standard solution A

CS  = concentration of USP Psoralen RS in Standard solution A (mg/mL)

CU  = concentration of Cullen corylifolium Fruit Dry Extract in the Sample solution (mg/mL)

 

Calculate the percentages of the labeled amount of psoralen in the portion of Cullen corylifolium Fruit Dry Extract taken:

 

Result = (P/L) × 100

 

P   = content of psoralen as determined above (%)

L   = labeled amount of psoralen (%)

 

Acceptance criteria: 90.0%–110.0% on the dried basis

 

CONTAMINANTS

• Elemental Impurities—Procedures <233>

Acceptance criteria

Arsenic: NMT 2.0 µg/g

Cadmium: NMT 0.5 µg/g

Lead: NMT 5.0 µg/g

Mercury: NMT 0.2 µg/g

• Articles of Botanical Origin <561>, Pesticide Residue Analysis: 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

• Loss on Drying <731>

Sample: 2 g of Cullen corylifolium Fruit Dry Extract 

Analysis: Dry the Sample at 105° for 5 h.

Acceptance criteria: NMT 8%

 

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 Cullen corylifolium Fruit Dry Extract RS

USP Psoralen RS

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Other Versions

Proposed For Comment Version 0.2