(nee' oh tame).
l-Phenylalanine, N-[N-(3,3-dimethylbutyl)-l--aspartyl]-1-methyl ester;
N-[N-(3,3-Dimethylbutyl)-l--aspartyl]-l-phenylalanine 1-methyl ester [165450-17-9].
Neotame contains NLT 97.0% and NMT 102.0% of neotame (C20H30N2O5), calculated on the anhydrous basis.
Mobile phase: Dissolve 3.0 g of sodium 1-heptanesulfonate in 740 mL of water in a suitable 1000-mL vessel, and add 3.8 mL of triethylamine. Adjust the resulting solution with phosphoric acid to a pH of 3.5, and dilute with water to 750 mL. Add 250 mL of acetonitrile, and adjust with phosphoric acid to an apparent pH of 3.7.
Standard solution: 1.0 mg/mL of USP Neotame RS in Mobile phase
Sample solution: 1.0 mg/mL of Neotame in Mobile phase.
[NoteThis solution is stable for up to 32 h when stored at a temperature of 010. ]
Detector: UV 210 nm
Column: 4.6-mm × 10-cm; packing L1
Column temperature: 45
Flow rate: 1.5 mL/min
Injection size: 25 µL
Sample: Standard solution
Tailing factor: NMT 2.0
Relative standard deviation: NMT 2.0%
Samples: Standard solution and Sample solution
Calculate the percentage of neotame (C20H30N2O5) in the portion of Neotame taken:
Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: 97.0%102.0% on the anhydrous basis
• Residue on Ignition 281: NMT 0.2%
• Lead 251
[NoteUse acid-cleaned (mixture of 5% nitric acid and 5% hydrochloric acid followed by rinsing with water) autosampler cups and volumetric glassware to avoid contamination. For the preparation of all aqueous solutions and for the rinsing of glassware before use, use water that has been passed through a strong-acid, strong-base, mixed-bed ion-exchange resin. Select all reagents to have as low a content of lead as practicable. Store standards and samples in acid-cleaned polyethylene containers. ]
Diluent: Transfer 2 mL of lead-free nitric acid into a 1000-mL volumetric flask, dilute with water to volume, and mix.
Standard stock solution: 79.9 mg of lead nitrate in 100 mL of Diluent in a 500-mL volumetric flask, and dilute with Diluent to volume. Transfer 10.0 mL of the resulting solution into a 100-mL volumetric flask, and dilute with Diluent to volume. Each mL of the Standard stock solution contains the equivalent of 10 µg of lead.
Standard solution A: Dilute an aliquot of the Standard stock solution with Diluent to obtain a solution having a concentration of 0.03 µg/mL.
Standard solution B: Dilute an aliquot of the Standard stock solution with Diluent to obtain a solution having a concentration of 0.015 µg/mL.
Sample solution: Transfer 160 mg of Neotame to a 10-mL volumetric flask. Dissolve in and dilute with Diluent to volume.
Mode: Atomic absorption spectrophotometer with a graphite furnace, pyrolytically coated graphite tubes, a solid pyrolytic graphite platform, and a background compensation system
Analytical wavelength: 283.3 nm
Lamp: Lead hollow-cathode
Purge gas: Argon
Alternate gas: Breathing-quality air
Volume: 15 µL. [NoteOptimize the instrument program as recommended by the manufacturer for lead, using a char temperature of 500 and an atomization temperature of 2000. ]
Samples: Standard solution A, Standard solution B, Sample solution, and Blank
Correct the area responses of the Sample solution, Standard solution A, and Standard solution B for the Blank area response. Generate the appropriate lead calibration algorithm, and determine the lead concentration in the Sample solution, in µg/mL.
Calculate the ppm of lead in the portion of Neotame taken:
Result = (C/W) × V × F
Acceptance criteria: NMT 2 ppm
• Related Compounds
Mobile phase and Chromatographic system: Proceed as directed in the Assay.
Standard solution A: 0.03 mg/mL of USP Neotame Related Compound A RS in Mobile phase
Standard solution B: Prepare as directed for the Standard solution in the Assay.
Detector sensitivity solution: Transfer 2 mL of Standard solution A to a 50-mL volumetric flask, and dilute with Mobile phase to volume.
Sample solution: 2 mg/mL of Neotame in Mobile phase.
[NoteThis solution is stable for up to 32 h when stored at a temperature of 0 to 10. ]
Samples: Standard solution A and Detector sensitivity solution
Signal-to-noise ratio: NLT 10, Detector sensitivity solution
Relative standard deviation: NMT 5.0%, Standard solution A
Samples: Standard solution A, Standard solution B, and Sample solution
Calculate the percentage of neotame related compound A in the portion of Neotame taken:
Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: NMT 1.5%
Calculate the percentage of other impurities in the portion of Neotame taken:
Result = (rT/rS) × (CS/CU) × 100
Acceptance criteria: NMT 2.0%
• Optical Rotation, Specific Rotation 781S: 40.0 to 43.4, at 20
Sample solution: 5 mg/mL in water
• Water Determination, Method Ic 921: NMT 5.0%
• Packaging and Storage: Preserve in well-closed containers, store in a dry place, and avoid exposure to excessive heat.
• USP Reference Standards 11
USP Neotame RS
USP Neotame Related Compound A RS
Auxiliary Information Please check for your question in the FAQs before contacting USP.
USP35NF30 Page 1874Pharmacopeial Forum: Volume No. 31(2) Page 497