Adenosine CAS 58-61-7 Assay 99.0%-101.0% USP Standard Factory High Purity

DEFINITION
Adenosine contains NLT 99.0% and NMT 101.0% of C10H13N5O4, calculated on the dried basis.
IDENTIFICATION
• A. INFRARED ABSORPTION <197K>: NMT 0.1%
ASSAY
• ADENOSINE
Sample: 200 mg of Adenosine previously dried at 105° for 2h
Titrimetric system
(See Titrimetry <541>)
Mode: Direct titration
Titrant: 0.1 N per chloric acid VS
Endpoint detection: Potentiometric
Analysis: Dissolve in 50 mL of glacial acetic acid and titrate with 0.1 N per chloric acid VS. Calculate the per centage of Adenosine (C10H13N5O4) in the portion taken:
Result = [(V − B) × N × F × 100]/W
V = Sample titrant volume (mL)
B = Blank titrant volume (mL)
N= titrant normality (mEq/mL)
F= equivalency factor: 267.25 mg/mEq
W= weight of the Sample (mg)
Acceptance criteria: 99.0%-101.0% on the dried basis
• IMPURITIES
• RESIDUEON IGNITION <281>: NMT 0.1%
• HEAVY METALS, Method II <231>: NMT 10 ppm
• LIMIT OF AMMONIA
Sample solution: Suspend 0.5 g in 10 mL of water. Stir for 30 s, and pass through a coarse filter. Dilute the filtrate with water to 15 mL, and use the filtrate.
Standard solution: 0.4 µg/mL of ammonium chloride in water
Analysis: To the Sample solution and the Standard solution add 0.3 mL of alkaline mer curic–potassium iodide TS, cap the test tubes, and allow to stand for 5 min.
Acceptance criteria: The Sample solution does not exhibit a more intense yellow color than that of the Standard solution (NMT 4 ppm of ammonia).
• LIMIT OF CHLORIDE
Sample solution: Suspend 0.2 g in 10 mL of water. Stir for 30 s, pass through a coarse filter, and use the filtrate.
Standard solution: 2.3 µg/mL of sodium chloride in water
Analysis: To the Sample solution and 10 mL of the Standard solution add 1 mL of nitric acid and 1 mL of silver nitrate TS, and dilute each solution with water to 40 mL. Allow the solutions to stand for 5 min, protected from light.
Acceptance criteria: When viewed against a dark background, the Sample solution is not more turbid than the Standard solution (NMT 0.007% chloride).
• LIMIT OF SULFATE
Sample solution: Suspend 0.75 g in 15 mL of water. Stir for 30 s, pass through a coarse filter, and use the filtrate.
Standard solution: Add 0.15 mL of 0.020 N sulfuric acid to 15 mL of water.
Analysis: To the Sample solution and the Standard solution add 2 mL of barium chloride TS and 1 mL of 3 N hydrochloric acid, dilute each solution with water to 30 mL, and mix. Allow the solutions to stand for 5 min.
Acceptance criteria: The Sample solution is not more turbid than the Standard solution (NMT 0.02% sulfate).
• ORGANIC IMPURITIES
Solution A: 6.8 g/L of potassium hydrogen sulfate and 3.4g/L of tetrabutylammonium hydrogen sulfate in water. Adjust with 2 N potassium hydroxide to a pH of 6.5.
Solution B: 0.1 g/L of sodium azide solution
Mobile phase: Solution A and Solution B (60:40)
System suitability solution: 0.2 mg/mL each of Adenosine and inosine in Mobile phase
Sample solution: 1.0 mg/mL of Adenosine in Mobile phase
Chromatographic system
(See Chromatography <621>, System Suitability.)
Mode: LC
Detector: UV 254 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Flow rate: 1.5 mL/min
Injection size: 20 µL
System suitability
Samples: System suitability solution
Suitability requirements
Resolution: NLT 9.0 between adenosine and inosine
Tailing factor: NMT 2.5
Relative standard deviation: NMT 2.0%
[NOTE-Chromatograph the Sample solution, and adjust the run time to at least twice the retention time of the major peak.]
Analysis
Sample: Sample solution
Calculate the percentage of each impurity in the portion of Adenosine taken:
Result = (rU/rT) × 100
rU = peak response of each impurity from the Sample solution
rT = sum of all the responses for all peaks from the Sample solution
Acceptance criteria
Individual impurities: NMT 0.1% each of guanosine, inosine, and uridine, and NMT 0.2% of adenine
Total impurities: NMT 0.5%
SPECIFIC TESTS
• MELTING RANGE OR TEMPERATURE <741>: 233°-238°
• OPTICAL ROTATION, Specific Rotation <781S>: -68° to -72°
Test solution: 20 mg/mL in sodium hydroxide solution (1 in 20), determined on a sample previously dried at 105° for 2h
ACIDITY OR ALKALINITY: Suspend 1 g in 20 mL of carbon dioxide-free water. Stir for 30 s, and pass through a coarse filter. To each of two 10-mL portions of the filtrate add 0.1mL of bromocresol purple TS.
Acceptance criteria: NMT 0.3 mL of 0.01 N sodium hydroxide is required to produce a blue-violet color in one portion. NMT 0.1 mL of 0.01 N hydrochloric acid is required to produce a yellow color in the other portion.
•LOSSON DRYING <731>: Dry a sample at 105℃ for 2 h: it loses NMT 0.5% of its weight.
ADDITIONAL REQUIREMENTS
•PACKAGING AND STORAGE: Preserve in tight, light-resistant containers, and store at controlled room temperature.
• USP REFERENCE STANDARDS <11>
USP Adenosine RS