S-Adenosyl-l-methionine Disulfate Tosylate Former Title: Ademetionine Disulfate Tosylate C22H34N6O16S4766.80 S-(Adenosyl)-l-methionine disulfate tosylate; (3S)-5¢-[(3-Amino-3-carboxypropyl)methylsulfonio]-5¢-deoxyadenosine, disulfate-methylbenzenesulfonate [97540-22-2]. DEFINITION S-Adenosyl-l-methionine Disulfate Tosylate is the disulfatetosylate mixed salt of a mixture of diastereoisomers of the S-adenosyl-l-methionine ion. It contains NLT 95.0% and NMT 105.0% of S-adenosyl-l-methionine disulfate tosylate (C22H34N6O16S4) calculated through the content of S-adenosyl-l-methionine (C15H23N6O5S+), calculated on the anhydrous basis. IDENTIFICATION • B. The retention time of the major peak of the Sample solution corresponds to that of S-adenosyl-l-methionine in the System suitability solution, as obtained in the Content of S-Adenosyl-l-Methionine. COMPOSITION • Content of S-Adenosyl-l-methionine Solution A: 10 mL of glacial acetic acid in 500 mL of water. Add 2.06 g of sodium 1-hexanesulfonate, and dilute with water to 1000 mL. Mobile phase: Acetronitrile and Solution A (15:85) System suitability solution: 400 µg/mL each of USP S-Adenosyl-l-methionine Disulfate Tosylate RS and USP S-Adenosyl-l-homocysteine RS Standard solution A: 400 µg/mL of USP S-Adenosyl-l-homocysteine RS Standard solution B: 200 µg/mL from Standard solution A Standard solution C: 80 µg/mL from Standard solution A Sample solution: 20 mg of S-Adenosyl-l-methionine Disulfate Tosylate in 40 mL of water. Stir for 30 min, then dilute with water to 50.0 mL. Transfer 1.0 mL of the solution to a 1.5-mL microcentrifuge tube, and centrifuge for 1 min. Use a portion of the supernatant. Chromatographic system Mode: LC Detector: UV 260 nm Column: 4.6-mm × 15-cm; 3-µm packing L1 Flow rate: 1 mL/min Injection size: 10 µL System suitability Samples: System suitability solution and Standard solution B [NoteThe relative retention times for S-adenosyl-l-homocysteine and S-adenosyl-l-methionine disulfate tosylate are about 0.68 and 1.0, respectively. ] Suitability requirements Resolution: NLT 1.5 between S-adenosyl-l-homocysteine and S-adenosyl-l-methionine Tailing factor: NMT 1.5, Standard solution B Relative standard deviation: NMT 2.0% for S-adenosyl-l-homocysteine, Standard solution B Analysis Samples: Standard solution A, Standard solution B, Standard solution C, and Sample solution [NoteRecord the chromatograms, and measure the area of the S-adenosyl-l-homocysteine peak in all three Standard solutions and the S-adenosyl-l-methionine disulfate tosylate peak in the Sample solution. ] Plot a calibration curve of the peak area of the Standard solutions versus the corresponding S-adenosyl-l-homocysteine concentration, in mg/mL, and draw the straight line best fitting the three points. From the calibration curve, and using the peak area of S-adenosyl-l-methionine from the chromatogram from the Sample solution, determine the concentration, C, in mg/mL, of S-adenosyl-l-methionine as S-adenosyl-l-homocysteine in the Sample solution. Calculate the percentage of C15H23N6O5S+ in the portion of S-Adenosyl-l-methionine Disulfate Tosylate taken: Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100
Acceptance criteria: 95.0%105.0% on the anhydrous basis • Content of Sulfate Mobile phase: 8.0 mM sodium carbonate and 1.0 mM sodium bicarbonate in water Standard solution: 0.18 mg/mL of potassium sulfate Sample solution: 0.5 mg/mL of S-Adenosyl-l-methionine Disulfate Tosylate Chromatographic system Mode: LC Detector: Ion detector with suppressed conductivity Column: 4.0-mm × 25-cm; 7-µm packing L74 Column temperature: 30 Flow rate: 1 mL/min Injection size: 25 µL System suitability Sample: Standard solution Suitability requirements Column efficiency: NLT 8200 theoretical plates Tailing factor: NMT 1.5 Relative standard deviation: NMT 1.0% Analysis Samples: Standard solution and Sample solution [NoteMeasure the area of the sulfate peak. ] Calculate the percentage of sulfate (SO4) in the portion of S-Adenosyl-l-methionine Disulfate Tosylate taken: Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: 23.5%26.5% IMPURITIES • Heavy Metals, Method I 231: NMT 20 ppm SPECIFIC TESTS • pH 791: 1.02.0, in an aqueous solution (1 in 20) • Water Determination, Method Ia 921: NMT 3.0% • Isomeric Ratio Buffer: Transfer 4.2 g of citric acid monohydrate and 2.03 g of sodium dihydrogen phosphate dihydrate to a 1-L volumetric flask, and dissolve in and dilute with water to volume. Mobile phase: 4.0 g of sodium dodecyl sulfate and 440 mL of acetonitrile. Dilute with Buffer to 1 L. Standard solution: 1.0 mg/mL of USP S-Adenosyl-l-methionine Disulfate Tosylate RS Sample solution: 1.0 mg/mL of S-Adenosyl-l-methionine Disulfate Tosylate Chromatographic system Mode: LC Detector: UV 254 nm Column: 4.6-mm × 25-cm; 5-µm packing L1 Flow rate: 1.2 mL/min Injection size: 20 µL System suitability Sample: Standard solution [NoteThe relative retention times for R,S-isomers and S,S-isomers are about 0.94 and 1.0, respectively. ] Suitability requirements Resolution: NLT 1.0 between the S,S-isomer and the R,S-isomer Analysis Samples: Standard solution and Sample solution Identify the peaks of the S,S- and R,S-isomers of the Sample solution by comparison with the Standard solution, and calculate the percentage of the S,S-isomer: Result = [rSS/(rSS + rRS)] × 100
Acceptance criteria: NLT 60% and NLT the labeled amount of the S,S-isomer ADDITIONAL REQUIREMENTS • Packaging and Storage: Preserve in tight, light-resistant containers, and store in a refrigerator. • Labeling: Label it to indicate the minimum content of S,S-isomer, as a percentage. • USP Reference Standards 11 USP S-Adenosyl-l-methionine Disulfate Tosylate RS USP S-Adenosyl-l-homocysteine RS Auxiliary Information Please check for your question in the FAQs before contacting USP.
USP35NF30 Page 1424 Pharmacopeial Forum: Volume No. 36(2) Page 438Chromatographic Column Chromatographic columns text is not derived from, and not part of, USP 35 or NF 30. |