1/8/2019 USP-NF

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Iohexol
To view the Notice from the Expert Committee that posted in conjunction with this accelerated revision, please click https://www.uspnf.com/rb- iohexol-20190726

C19H26I3N3O9 821.14
1,3-Benzenedicarboxamide, 5-[acetyl(2,3-dihydroxypropyl)amino]-N,N′-bis(2,3-dihydroxypropyl)-2,4,6-triiodo;
N,N′-Bis(2,3-dihydroxypropyl)-5-[N-(2,3-dihydroxypropyl)acetamido]-2,4,6-triiodoisophthalamide [66108-95-0]; UNII: 4419T9MX03.

DEFINITION
Iohexol contains NLT 98.0% and NMT 102.0% of iohexol (C19H26I3N3O9), calculated on the anhydrous basis.

IDENTIFICATION
• A. INFRARED ABSORPTION 〈197K〉
• B. The retention times of the two principal peaks of the Sample solution correspond to those of the System suitability solution, as obtained in the test for Organic
Impurities.

ASSAY
• PROCEDURE
Sample: 500 mg of Iohexol
Sample solution: Transfer the Sample to a glass-stoppered, 125-mL conical ask. Add 25 mL of 1.25 N sodium hydroxide and 500 mg of powdered zinc. Connect th
ask to a re ux condenser, and re ux for 1 h. Cool the ask to room temperature, rinse the condenser with 20 mL of water, disconnect the ask from the
condenser, and pass the mixture through a lter. Rinse the ask and the lter thoroughly with small portions of water, adding the rinsings to the ltrate. Add 5 mL
of glacial acetic acid.
Titrimetric system
(See Titrimetry 〈541〉.)
Mode: Direct titration
Titrant: 0.1 N silver nitrate VS
Endpoint detection: Potentiometric
Analysis: Titrate the Sample solution with 0.1 N silver nitrate VS.
Calculate the percentage of iohexol (C19H26I3N3O9) in the portion of Iohexol taken:

Result = [(V × N × F)/W] × 100

V = Titrant volume consumed by the Sample (mL)

N = Titrant normality (mEq/mL)

F = equivalent weight of iohexol, 273.7 mg/mEq

W = Sample weight (mg)

Acceptance criteria: 98.0%–102.0% on the anhydrous basis

IMPURITIES
• LIMIT OF IONIC COMPOUNDS
Rinse all glassware ve times with distilled water.
Standard solution: 0.002 mg/mL of sodium chloride in water
Sample solution: 1 g of Iohexol in 50 mL of water
Analysis
Samples: Standard solution and Sample solution

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Acceptance criteria: The speci c conductance of the Sample solution is NMT that of the Standard solution (equivalent to 0.01% ionic compounds as sodium
chloride).
• LIMIT OF FREE IODIDE
Sample: 5 g of Iohexol
Sample solution: Dissolve the Sample in 20 mL of water.
Titrimetric system
(See Titrimetry 〈541〉.)
Mode: Direct titration
Titrant: 0.001 N silver nitrate VS
Endpoint detection: Potentiometric
Analysis
Calculate the percentage of free iodide in the portion of the Sample taken:

Result = [(V × N × F)/W] × 100

V = Titrant volume consumed by the Sample (mL)

N = Titrant normality (mEq/mL)

F = equivalent weight of iodide, 126.9 mg/mEq

W = Sample weight (mg)

Acceptance criteria: NMT 0.001%

Change to read:
• ORGANIC IMPURITIES
Solution A: Acetonitrile
Solution B: Water
Mobile phase: See Table 1.

Table 1

Time (min) Solution A (%) Solution B (%)

0 1 99

60 13 87

System suitability solution: 1.5 mg/mL of USP Iohexol RS and 0.0075 mg/mL of USP Iohexol Related Compound A RS in water
Sample solution: 1.5 mg/mL of Iohexol
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC
Detector: UV 254 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Flow rate: 1 mL/min
Injection volume: 10 µL
System suitability
Sample: System suitability solution
[NOTE— Iohexol may give two nonresolved peaks due to exo–endo isomerism. In addition, a small peak due to iohexol usually appears at the leading edge of the
rst principal peak. This small peak has a retention time about 1.2 min less than the rst principal peak. The relative retention times for the iohexol related
compound A, iohexol endo-isomer, iohexol exo-isomer, and O-alkylated compounds peaks are 0.85, 0.96, 1.0, and 1.1–1.4, respectively.]
Suitability requirements
Resolution: NLT 5.0 between iohexol related compound A and the exo-isomer (the second and greater peak) of iohexol
Analysis
Sample: Sample solution
Calculate the percentage of O-alkylated compounds and any other individual impurity in the portion of Iohexol taken.
▲Exclude peaks with a relative retention time between 0.84 [relative to the endo-isomer of iohexol ( rst main peak)] and that of the endo-isomer of iohexol.▲

(RB 1-Aug-2019)

Disregard any peak less than or equal to 0.03% of the principal peaks.

Result = (rU/rT) × 100

rU = peak response of each impurity

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rT = sum of all of the peak responses

Acceptance criteria
O-alkylated compounds: NMT 0.6%
Any individual impurity: NMT 0.1%
Total impurities excluding O-alkylated compounds: NMT 0.3%
• LIMIT OF 2-METHOXYETHANOL
Internal standard solution: 0.01 mg/mL of secondary butyl alcohol in water
Standard stock solution: 0.005 mg/mL of methanol and 0.01 mg/mL each of isopropyl alcohol, secondary butyl alcohol, and 2-methoxyethanol in Internal standard
solution
Standard solution: Transfer about 0.25 g of USP Iohexol RS and 1.0 mL of Standard stock solution to a headspace vial, and seal the vial with a septum and crimp
cap.
Sample solution: Transfer about 0.25 g of Iohexol and 1.0 mL of Internal standard solution to a headspace vial, and seal the vial with a septum and crimp cap.
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: GC with suitable headspace autosampler
Detector: Flame ionization
Column: 0.53-mm × 30-m fused-silica coated with a 1-µm phase G16
Temperatures
Autosampler: 105°
Needle: 130°–140°
Injection port: 150°
Detector: 200°
Column: See Table 2.

Table 2

Hold Time at Final Temperature

Initial Temperature (°) Temperature Ramp (°/min) Final Temperature (°) (min)

40 — 40 3

40 8 100 1

Carrier gas: Helium

Flow rate: 11 mL/min
Injection volume: 1 mL of the headspace
System suitability
Sample: Standard solution
[NOTE—The typical relative retention times for methanol, isopropyl alcohol, secondary butyl alcohol, and 2-methoxyethanol are 0.5, 0.6, 1.0, and 1.9, respectively.]
Suitability requirements
Resolution: NLT 1.0 between methanol and isopropyl alcohol
Relative standard deviation: NMT 10.0% for the ratio of 2-methoxyethanol to the internal standard
Analysis
Samples: Standard solution and Sample solution
Calculate the amount of 2-methoxyethanol in the portion of Iohexol taken:

Result = (RU/RS) × (Cs/CU)

RU = peak response ratio of 2-methoxyethanol to the internal standard from the Sample solution

RS = peak response ratio of 2-methoxyethanol to the internal standard from the Standard solution

CS = concentration of 2-methoxyethanol in the Standard solution (µg/mL)

CU = concentration of Iohexol in the Sample solution (g/mL)

Acceptance criteria: NMT 20 µg/g of 2-methoxyethanol

• LIMIT OF 3-CHLOROPROPANE-1,2-DIOL
Standard solution: 0.025 mg/mL of 3-chloropropane-1,2-diol in ethyl acetate
Sample solution: Transfer 1 g of Iohexol to a separator. Dissolve in 1 mL of water. Extract 4 times with 2 mL of ethyl acetate, and combine the extracts. Dry the
combined extracts with anhydrous sodium sulfate. Filter, and wash the lter with a small amount of ethyl acetate. Combine the wash with the ltrate, and
concentrate to a volume of 0.7 mL, using a warm water bath and a stream of nitrogen. Dilute with ethyl acetate to 2 mL.
Chromatographic system

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(See Chromatography 〈621〉, System Suitability.)
Mode: GC
Detector: Flame ionization
Column: 0.32-mm × 30-m fused-silica capillary bonded with a 1-µm layer of phase G46
Temperatures
Injection port: 230°
Detector: 250°
Column: See Table 3.

Table 3

Hold Time at Final Temperature

Initial Temperature (°) Temperature Ramp (°/min) Final Temperature (°) (min)

80 — 80 2

80 15 275 —

275 — 275 2

Carrier gas: Helium at 1 mL/min

Injection volume: 2 µL
System suitability
Sample: Standard solution
[NOTE—The retention time of the 3-chloropropane-1,2-diol peak is about 8 min.]
Suitability requirements
Relative standard deviation: NMT 10.0%
Analysis
Samples: Standard solution and Sample solution
Acceptance criteria: The area of the principal peak from the Sample solution is NMT the area of the principal peak from the Standard solution (NMT 0.0025%).
• LIMIT OF FREE AROMATIC AMINE
Solution A: 3 mg/mL of N-(1-naphthyl)ethylenediamine dihydrochloride in a mixture of propylene glycol and water (70:30)
Standard stock solution: 10 µg/mL of USP Iohexol Related Compound B RS in water
Standard solution: Transfer 5 mL of water and 10.0 mL of the Standard stock solution to a 25-mL volumetric ask.
Sample solution: Transfer 200 mg of Iohexol to a 25-mL volumetric ask, add 15 mL of water, and mix to dissolve.
Blank: Add 15 mL of water to a 25-mL volumetric ask.
Instrumental conditions
Mode: Vis
Analytical wavelength: 495 nm
Cell: 5 cm
Analysis
Samples: Standard solution, Sample solution, and Blank
In conducting the following steps, keep the asks in iced water and protected as much as possible from light until all of the reagents have been added.
Treat the Samples as follows. Place the ask in an ice bath for 5 min. Add 1.5 mL of 6 N hydrochloric acid, and mix by swirling. Add 1.0 mL of sodium nitrite
solution (20 mg/mL), and allow to stand in the ice bath for 4 min. Remove the ask from the ice bath, add 1.0 mL of sulfamic acid solution (40 mg/mL), and
swirl gently until gas evolution ceases. [CAUTION—Considerable pressure is produced.] Add 1.0 mL of Solution A, dilute with water to volume, and allow to stan
for 5 min. Measure the absorbance of the Standard solution and Sample solution against the Blank solution.
Acceptance criteria: The absorbance of the Sample solution is NMT that of the Standard solution (NMT 0.05% of free aromatic amine).

SPECIFIC TESTS
• COLOR OF SOLUTION
Sample solution: 647.2 mg/mL
Blank: Water
Instrumental conditions
Mode: UV-Vis
Analytical wavelengths: 400, 420, and 450 nm
Cell: 1 cm
Analysis
Samples: Sample solution and Blank
Pass the Sample solution through a lter of 0.22-µm pore size.
Determine the absorbances of the Sample solution against the Blank.
Acceptance criteria: See Table 4.

Table 4

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Wavelength (nm) NMT (au)

400 0.180

420 0.030

450 0.015

• WATER DETERMINATION 〈921〉, Method I: NMT 4.0%

ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in well-closed, light-resistant containers. Store at room temperature.
• USP REFERENCE STANDARDS 〈11〉
USP Iohexol RS
USP Iohexol Related Compound A RS
5-(Acetylamino)-N,N′-bis(2,3-dihydroxypropyl)-2,4,6-triiodo-1,3-benzenedicarboxamide.
C16H20I3N3O7 747.06
USP Iohexol Related Compound B RS
5-Amino-N,N′-bis(2,3-dihydroxypropyl)-2,4,6-triiodo-1,3-benzenedicarboxamide.
C14H18I3N3O6 705.02

Auxiliary Information- Please check for your question in the FAQs before contacting USP.

Topic/Question Contact Expert Committee

IOHEXOL Ravi Ravichandran CHM42015 Chemical Medicines Monographs 4

Principal Scienti c Liaison
+1 (301) 816-8330

Chromatographic Columns Information: Chromatographic Columns

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