Printed on: Wed Feb 08 2023, 11:15:37 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-443A9951-3E44-46EC-A62B-2C0963DD591C_4_en-US

Printed by: Dang Van Vu Official Date: Official as of 01-May-2020 Document Type: NF @2023 USPC
Do Not Distribute DOI Ref: 4jcq1 DOI: https://doi.org/10.31003/USPNF_M78575_04_01
1

Standard solution: 0.2 µL/mL of methanol in Internal

Sucralose standard solution
Sample solution: 0.2 g/mL of Sucralose in Internal standard
solution
Chromatographic system
(See Chromatography á621ñ, System Suitability.)
Mode: GC
Detector: Flame ionization
C12H19Cl3O8 397.63 Column: 4-mm × 2-m glass column; packed with 80- to
1,6-Dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4- 100-mesh silanized support S6
deoxy-α-D-galactopyranoside; Temperature
1′,4,6′-Trichlorogalactosucrose CAS RN®: 56038-13-2. Column: 150°
Detector: 250°
DEFINITION Injector: 200°
Sucralose contains NLT 98.0% and NMT 102.0% Carrier gas: Helium
of C12H19Cl3O8, calculated on the anhydrous basis. Flow rate: 20 mL/min
Injection size: 1 µL
IDENTIFICATION System suitability
Change to read: Sample: Standard solution
Suitability requirements
• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Relative standard deviation: NMT 2.0%
Spectroscopy: 197K▲ (CN 1-May-2020) Analysis

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• B. The retention time of the principal peak of the Sample Samples: Standard solution and Sample solution
solution corresponds to that of the Standard solution, as Calculate the percentage of methanol in the portion of
obtained in the Assay. Sucralose taken:
• C. The R F value of the principal spot of the Sample solution
corresponds to that of Standard solution A, as obtained in Result = (R U/R S) × [(C S/C U) × F 1] × F 2 × 100
the test for Related Compounds.
ASSAY
• PROCEDURE
ci RU

RS
= peak response ratio of methanol to n-propyl
alcohol, from the Sample solution
= peak response ratio of methanol to n-propyl
Mobile phase: Acetonitrile and water (3:17) alcohol, from the Standard solution
Standard solution: 1 mg/mL of USP Sucralose RS in Mobile CS = concentration of methanol in the Standard
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phase solution (µL/mL)
Sample solution: 1 mg/mL of Sucralose in Mobile phase CU = concentration of Sucralose in the Sample solution
Chromatographic system (g/mL)
(See Chromatography á621ñ, System Suitability.) F1 = conversion factor from µL to mL
Mode: LC F2 = specific gravity of methanol, 0.79 g/cm3
Detector: Refractive index
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Column: 8-mm × 10-cm; packing L1 Acceptance criteria: NMT 0.1%

Flow rate: 1.5 mL/min • RELATED COMPOUNDS
Injection size: 20 µL Adsorbent: 0.20-mm layer of octadecylsilanized
System suitability chromatographic silica gel. The thin-layer chromatographic
Sample: Standard solution plate also has a preadsorbent zone.
[NOTE—The retention time of sucralose is about 9 min.] Detection reagent: Sulfuric acid in methanol (3 in 20)
Suitability requirements Standard solution A: 10.0 mg/mL of USP Sucralose RS in
Relative standard deviation: NMT 2.0% methanol
Analysis Standard solution B: 0.5 mL Standard solution A diluted to
Samples: Standard solution and Sample solution 10.0 mL with methanol
Calculate the percentage of sucralose (C12H19Cl3O8) in the Sample solution: 100.0 mg/mL of Sucralose in methanol
portion of Sucralose taken: Developing solvent system: Acetonitrile and sodium
chloride solution (1 in 20) (3:7)
Result = (r U/r S) × (C S/C U) × 100 Application volume: 5 µL
Analysis
rU = peak response of the Sample solution Samples: Standard solution A, Standard solution B, and
rS = peak response of the Standard solution Sample solution
CS = concentration of USP Sucralose RS in the Standard Proceed as directed under Chromatography á621ñ,
solution (mg/mL) Thin-Layer Chromatography. Spray the plate with Detection
CU = concentration of Sucralose in the Sample solution reagent. Heat the plate for 10 min at 125°.
(mg/mL) Acceptance criteria: The R F value of the principal spot from
the Sample solution corresponds to that obtained from
Acceptance criteria: 98.0%–102.0% on the anhydrous Standard solution A, and the color of any other single spot
basis from the Sample solution is not more intense than that of
IMPURITIES the principal spot from Standard solution B (0.5%).
• RESIDUE ON IGNITION á281ñ: NMT 0.7% • LIMIT OF HYDROLYSIS PRODUCTS
• LIMIT OF METHANOL [NOTE—This test does not require a developing
Internal standard solution: 0.1 µL/mL of n-propyl alcohol solvent.]
in pyridine Adsorbent: 0.25-mm layer of chromatographic silica gel

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Printed on: Wed Feb 08 2023, 11:15:37 PM(EST) Status: Currently Official on 09-Feb-2023 DocId: GUID-443A9951-3E44-46EC-A62B-2C0963DD591C_4_en-US
Printed by: Dang Van Vu Official Date: Official as of 01-May-2020 Document Type: NF @2023 USPC
Do Not Distribute DOI Ref: 4jcq1 DOI: https://doi.org/10.31003/USPNF_M78575_04_01
2

Spray reagent: 12.3 mg/mL of p-anisidine and 16.6 mg/mL test, heating for a shorter period of time. Immediately
of phthalic acid in methanol. Store the solution in the dark after heating, view the plate against a dark background.
and refrigerate to prevent discoloration. Discard if the Acceptance criteria: The color of the spot from the Sample
solution becomes discolored. [CAUTION—p-Anisidine is toxic solution is not more intense than that from Standard solution
if inhaled or if absorbed through the skin.] B (0.1%).
Standard solution A: 100 mg/mL of mannitol
Standard solution B: 0.4 mg/mL of fructose and SPECIFIC TESTS
100 mg/mL of mannitol • OPTICAL ROTATION, Specific Rotation á781Sñ: +84.0° to
Sample solution: 250 mg/mL of Sucralose in methanol +87.5° at 20°
Application volume: 5-µL portions separately applied in Sample solution: 10 mg/mL of Sucralose
1-µL increments, allowing the plate to dry between • WATER DETERMINATION, Method I á921ñ: NMT 2.0%
applications ADDITIONAL REQUIREMENTS
Analysis • PACKAGING AND STORAGE: Preserve in well-closed
Samples: Standard solution and Sample solution containers, in a cool, dry place, at a temperature not
Proceed as directed under Chromatography á621ñ, exceeding 21°.
Thin-Layer Chromatography. Spray the plate with Spray • USP REFERENCE STANDARDS á11ñ
reagent, and heat the plate at 100 ± 2° for 15 min. If the USP Sucralose RS
spot from Standard solution A has darkened, repeat the

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