(a den' oh seen).
Adenosine contains NLT 99.0% and NMT 101.0% of C10H13N5O4, calculated on the dried basis.
• Infrared Absorption 197M: NMT 0.1%
Sample: 200 mg of Adenosine previously dried at 105 for 2 h
(See Titrimetry 541.)
Mode: Direct titration
Titrant: 0.1 N perchloric acid VS
Endpoint detection: Potentiometric
Blank: 50 mL of glacial acetic acid
Analysis: Dissolve in 50 mL of glacial acetic acid and titrate with 0.1 N perchloric acid VS. Calculate the percentage of Adenosine (C10H13N5O4) in the portion taken:
Result = [(V B) × N × F × 100]/W
Acceptance criteria: 99.0%101.0% on the dried basis
• Residue on 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 mercuricpotassium 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 3.4 g/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
Detector: UV 254 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Flow rate: 1.5 mL/min
Injection size: 20 µL
Sample: System suitability solution
Resolution: NLT 9.0 between adenosine and inosine
Tailing factor: NMT 2.5
Relative standard deviation: NMT 2.0%
[NoteChromatograph the Sample solution, and adjust the run time to at least twice the retention time of the major peak. ]
Sample: Sample solution
Calculate the percentage of each impurity in the portion of Adenosine taken:
Result = (rU/rT) × 100
Individual impurities: NMT 0.1% each of guanosine, inosine, and uridine, and NMT 0.2% of adenine
Total impurities: NMT 0.5%
• Melting Range or Temperature 741: 233238
• 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 2 h
• 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 0.1 mL 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.
• Loss on Drying 731: Dry a sample at 105 for 2 h: it loses NMT 0.5% of its weight.
• Packaging and Storage: Preserve in tight, light-resistant containers, and store at controlled room temperature.
• USP Reference Standards 11
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USP35NF30 Page 2079Pharmacopeial Forum: Volume No. 29(6) Page 1834