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1 by ebsc0sa on Mon Mar 28 12:07:37 EDT 2016

NF 34

Official Monographs / Aspartame 7165

SPECIFIC TESTS
OPTICAL ROTATION, Specific Rotation 781S
Sample solution: 10 mg/mL, in 6 N hydrochloric acid
Acceptance criteria: +33.0 to +36.5, measured at 20
MICROBIAL ENUMERATION TESTS 61 and TESTS FOR SPECIFIED MICROORGANISMS 62: The total aerobic microbial
count does not exceed 1000 cfu/g, and the total combined molds and yeasts count does not exceed 100 cfu/
g.
LOSS ON DRYING 731: Dry a sample at 130 for 3 h: the
anhydrous form loses NMT 1.0% of its weight, and the
monohydrate loses between 11.5% and 12.5% of its
weight.

Suitability requirements
Resolution: NLT 8 between the L-phenylalanine and
aspartame related compound A peaks, System suitability solution
Tailing factor: NMT 1.5, Standard solution
Relative standard deviation: NMT 1.0%, Standard
solution
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of aspartame (C14H18N2O5) in
the portion of Aspartame taken:

ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in well-closed, lightresistant containers. Store at room temperature.
LABELING: Label it to indicate whether it is anhydrous or
the monohydrate.
USP REFERENCE STANDARDS 11
USP Asparagine Anhydrous RS
USP Asparagine Monohydrate RS

rU

Aspartame

Result = (rU/rS) (CS/CU) 100

= peak area of aspartame from the Sample
solution
rS
= peak area of aspartame from the Standard
solution
CS
= concentration of USP Aspartame RS in the
Standard solution (mg/mL)
CU
= concentration of Aspartame in the Sample
solution (mg/mL)
Acceptance criteria: 98.0%102.0% on the dried basis
IMPURITIES
RESIDUE ON IGNITION 281: NMT 0.2%
Delete the following:

HEAVY METALS, Method II 231: NMT 10 ppm (Official 1.

Jan-2018)

C14H18N2O5
294.30
L-Phenylalanine, N-L--aspartyl-, 1-methyl ester;
3-Amino-N-(-carboxyphenethyl)succinamic acid N-methyl
ester [22839-47-0].
DEFINITION
Aspartame contains NLT 98.0% and NMT 102.0% of aspartame (C14H18N2O5), calculated on the dried basis.
IDENTIFICATION
A. INFRARED ABSORPTION 197M
[NOTEDo not dry specimens.]

Result = (rU/rS) (CS/CU) 100

rU

= peak response of aspartame related

compound A from the Sample solution
rS
= peak response of aspartame related
compound A from the Standard solution
CS
= concentration of 5-benzyl-3,6-dioxo2-piperazineacetic acid in the Standard
solution (mg/mL)
CU
= concentration of Aspartame in the Sample
solution (mg/mL)
Acceptance criteria: NMT 1.5%
CHROMATOGRAPHIC PURITY
Mobile phase, Diluent, System suitability solution, and
Chromatographic system: Proceed as directed in the
Assay.
Sample stock solution: Proceed as directed in the Sample solution in the test for Limit of 5-Benzyl-3,6-dioxo2-piperazineacetic Acid.

Official from May 1, 2016

Copyright (c) 2016 The United States Pharmacopeial Convention. All rights reserved.

NF Monographs

ASSAY
PROCEDURE
Buffer: 0.05 M monobasic potassium phosphate, adjusted with phosphoric acid to a pH of 4.3
Mobile phase: Methanol and Buffer (18:82)
Diluent: Methanol and water (1:9)
System suitability solution: 0.1 mg/mL each of USP
Aspartame Related Compound A RS and USP L-Phenylalanine RS in Diluent
Standard solution: 0.5 mg/mL of USP Aspartame RS in
Diluent
Sample solution: 0.5 mg/mL of Aspartame in Diluent.
[NOTEAvoid heat and excessive holding times.]
Chromatographic system
(See Chromatography 621, System Suitability.)
Mode: LC
Detector: UV 210 nm
Column: 4.6-mm 25-cm; 5-m packing L1
Column temperature: 40
Flow rate: 2 mL/min
Injection volume: 20 L
Run time: 30 min
System suitability
Samples: System suitability solution and Standard
solution
[NOTEThe relative retention times for the L-phenylalanine and aspartame related compound A peaks are 0.6
and 1.0, respectively.]

LIMIT OF 5-BENZYL-3,6-DIOXO-2-PIPERAZINEACETIC ACID

Mobile phase, Diluent, System suitability solution, and
Chromatographic system: Proceed as directed in the
Assay.
Standard solution: 75 g/mL of USP Aspartame Related
Compound A RS in Diluent
Sample solution: 5 mg/mL of Aspartame in Diluent.
[NOTEAvoid heat and excessive holding times.]
System suitability
Samples: System suitability solution and Standard
solution
Suitability requirements
Resolution: NLT 8 between the L-phenylalanine and
aspartame related compound A peaks, System suitability solution
Tailing factor: NMT 2.0, Standard solution
Relative standard deviation: NMT 4.0%, Standard
solution
Analysis
Samples: Standard solution and Sample solution
[NOTEContinue the elution of the Sample solution for
45 min.]
Calculate the percentage of aspartame related compound A in the portion of Aspartame taken:

Accessed from 10.6.1.1 by ebsc0sa on Mon Mar 28 12:07:37 EDT 2016

7166 Aspartame / Official Monographs

Sample solution: 0.1 mg/mL of Aspartame from the
Sample stock solution in Diluent
System suitability
Sample: System suitability solution
Suitability requirements
Resolution: NLT 8 between the L-phenylalanine and
aspartame related compound A peaks, System suitability solution
Analysis
Samples: Sample stock solution and Sample solution
[NOTEContinue the elution of the Sample stock solution for 45 min.]
Acceptance criteria: The sum of all of the peak responses of the Sample stock solution, excluding the
5-benzyl-3,6-dioxo-2-piperazineacetic acid and aspartame peak responses, is NMT the aspartame peak
response of the Sample solution, corresponding to NMT
2.0% of chromatographic impurities.
SPECIFIC TESTS
TRANSMITTANCE
Sample solution: 10 mg/mL of Aspartame in 2 N hydrochloric acid, prepared by means of sonication
Analysis: Determine the transmittance in a 1-cm cell at
430 nm with a suitable spectrophotometer.
Acceptance criteria: Transmittance of NLT 0.95, corresponding to an absorbance of NMT about 0.022
OPTICAL ROTATION, Specific Rotation 781S
Sample solution: 40 mg/mL in 15 N formic acid
Acceptance criteria: +14.5 to +16.5, determined at
20 within 30 min after preparation of the Sample
solution
LOSS ON DRYING 731
Analysis: Dry at 105 for 4 h.
Acceptance criteria: NMT 4.5%
ADDITIONAL REQUIREMENTS
PACKAGING AND STORAGE: Preserve in well-closed
containers.
USP REFERENCE STANDARDS 11
USP Aspartame RS
USP Aspartame Related Compound A RS
5-Benzyl-3,6-dioxo-2-piperazineacetic acid.
C13H14N2O4
262.27
USP L-Phenylalanine RS

NF Monographs

Aspartame Acesulfame

C18H23O9N3S
457.45
Aspartame acesulfame salt;
[2-Carboxy--(N-b-methoxycarbonyl-2-phenyl)ethylcarbamoyl)]ethanaminium 6-methyl-4-oxo-1,2,3-oxathiazin-3-ide-2,2-dioxide;
L-Phenylalanine, L--aspartyl-2-methyl ester compound with
6-methyl-1,2,3-oxathiazin-4(3H)-one 2,2-dioxide (1:1)
[106372-55-8].
DEFINITION
Aspartame Acesulfame contains NLT 63.0% and NMT
66.0% of aspartame, calculated on the dried basis. It contains NLT 34.0% and NMT 37.0% of acesulfame, calculated as the acid form on the dried basis.

NF 34
IDENTIFICATION
A. INFRARED ABSORPTION 197K
[NOTEDo not dry specimens.]
ASSAY
PROCEDURE
Sample: 0.150 g
Analysis: Dissolve the Sample in 50 mL of dehydrated
alcohol. Titrate with 0.1 N tetrabutylammonium hydroxide in methanol/isopropyl alcohol VS. Potentiometrically determine the volumes of titrant, in mL, at the
first equivalence point (V1) and at the second equivalence point (V2). Perform a blank titration with 50 mL of
dehydrated alcohol, and designate the volume of titrant, in mL, as VB.
Calculate the percentage of acesulfame (Result 1) and
aspartame (Result 2) in the portion of Aspartame
Acesulfame taken:
Result 1 = 163N(V1 VB)/10W
Result 2 = 294N(V2 V1)/10W
N
= normality of the titrant
W
= weight of Aspartame Acesulfame taken (g)
Acceptance criteria: Aspartame, 63.0%66.0% on the
dried basis; acesulfame, 34.0%37.0% as the acid form
on the dried basis
IMPURITIES
RESIDUE ON IGNITION 281: NMT 1.1%
Delete the following:

HEAVY METALS, Method II 231: NMT 10 ppm (Official 1.

Jan-2018)

Change to read:

LIMIT OF POTASSIUM
Potassium stock solution: 19.0 g/mL of potassium
chloride, previously dried at 105 for 2 h, in water. This
solution contains 10 g of potassium/mL.
Standard solution A: Transfer 10.0 mL of Potassium
stock solution to a 100-mL volumetric flask. Add 2.0 mL
of sodium chloride solution (1 in 5) and 1.0 mL of hydrochloric acid, and dilute with water to volume
(1.0 g/mL of potassium).
Standard solution B: Transfer 15.0 mL of Potassium
stock solution to a 100-mL volumetric flask. Add 2.0 mL
of sodium chloride solution (1 in 5) and 1.0 mL of hydrochloric acid, and dilute with water to volume
(1.5 g/mL of potassium).
Standard solution C: Transfer 20.0 mL of Potassium
stock solution to a 100-mL volumetric flask. Add 2.0 mL
of sodium chloride solution (1 in 5) and 1.0 mL of hydrochloric acid, and dilute with water to volume
(2.0 g/mL of potassium).
Sample stock solution: 3.0 mg/mL of Aspartame
Acesulfame
Sample solution: Transfer 10 mL of Sample stock solution to a 100-mL volumetric flask. Add 2.0 mL of sodium chloride solution (1 in 5) and 1.0 mL of hydrochloric acid, and dilute with water to volume. Filter the
solution.
Instrumental conditions
(See Atomic Absorption Spectroscopy 852 (CN 1-May2016).)
Mode: Atomic absorption spectrophotometry
Analytical wavelength: 766.5 nm (potassium emission
line)
.

Official from May 1, 2016

Copyright (c) 2016 The United States Pharmacopeial Convention. All rights reserved.