11/2/2020 USP-NF Iron Sucrose Injection

Printed on: Mon Nov 02 2020, 17:28:19 pm

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O cial Status: Currently O cial on 02-Nov-2020
O cial Date: O cial as of 1-May-2018
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© 2020 USPC

Iron Sucrose Injection

DEFINITION
Iron Sucrose Injection is a sterile, colloidal solution of ferric hydroxide in complex with Sucrose in Water for Injection. It contains NLT
95.0% and NMT 105.0% of the labeled amount of iron. Sodium Hydroxide may be added to adjust the pH. It contains no antimicrobial
agent, chelating agent, dextran, gluconate, or other added substances.

IDENTIFICATION
• A. IRON
To 2.5 mL of Injection add 17.5 mL of water and 5 mL of hydrochloric acid. Mix and heat the solution for 5 min in a boiling water bath.
Cool, add dropwise 13.5 N ammonium hydroxide until no further precipitation of ferric hydroxide occurs, and lter. Wash the
precipitate with water to remove excess ammonium hydroxide, dissolve the precipitate in a minimum volume of 2 N hydrochloric acid,
and add su cient water to make a volume of 20 mL. To 3 mL of the solution add 1 mL of 2 N hydrochloric acid and 1 mL of
potassium thiocyanate TS: the resulting solution (Solution 1) is red. To 1 mL of Solution 1 add 5 mL of amyl alcohol or ethyl ether,
shake, and allow to stand: the organic layer is pink. To a separate 1-mL aliquot of Solution 1 add 2 mL of mercuric chloride TS: a red
color is discharged [iron (III) salts].
• B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay
for Sucrose.
• C. MOLECULAR WEIGHT DETERMINATION
Mobile phase: Dissolve 7.12 g of dibasic sodium phosphate dihydrate, 5.52 g of monobasic sodium phosphate, and 0.40 g of sodium
azide in 2 L of water.

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System suitability solution: Dissolve 200 mg of high molecular weight dextran and 100 mg of glucose in 20 mL of Mobile phase.
Standard solutions: Transfer about 20 mg of each polysaccharide molecular weight standard (5,000–400,000 Da) to separate 5-mL
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volumetric asks. Add 4 mL of Mobile phase to each ask, and allow each aliquot to stand at or below 25° for a minimum of 12 h.
After the agglomerate particles of each Standard solution have swelled to their fullest extent, gently swirl each Standard solution
until dissolved.
[ NOTE— The chromatograms of freshly prepared Standard solutions regularly show a small, unidenti ed secondary peak following
the main peak. Discard the Standard solutions if the secondary peak reaches half the height of the main peak.]
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Sample solution: Transfer 5.0 mL of Injection to a 10-mL volumetric ask, and dilute with Mobile phase to volume.
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC
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Detector: Refractive index, maintained at a constant temperature of 45°

Columns: Two 7.8-mm × 30-cm; packing L39 with pore sizes of 1000 and 120 Å, respectively
Column temperature: 45 ± 2°
Flow rate: 0.5 mL/min
Injection volume: 25 µL
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System suitability
Samples: System suitability solution and Standard solutions
Suitability requirements
Resolution: NLT 4.0 between dextran and glucose, System suitability solution
Correlation coe cient: NLT 0.98 for the calibration curve generated using a suitable program, plotting the retention times of the
Standard solutions and their molecular weights to generate a third order (cubic) calibration curve
Analysis
Samples: System suitability solution, Standard solutions, and Sample solution
The molecular weight of the complex is calculated from the calibration curve. The molecular weight distribution curve of the
sample is sliced into fractions.
Calculate the weight-average molecular weight (MW) as follows:

Result = Σ(ATMT)/ΣAT
Calculate the number-average molecular weight (MN) as follows:

Result = Σ(AT)/Σ(AT/MT)

AT = area of each fraction of the sample distribution

MT = corresponding mean molecular weight of each fraction as determined from its retention time on the calibration curve

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Acceptance criteria: The molecular weight distribution curve of the Injection conforms to the following parameters.
MW: 34,000–60,000 Da
MN: NLT 24,000 Da
MW /MN: NMT 1.7

ASSAY
• SUCROSE
Mobile phase: Acetonitrile and water (79:21)
Standard solutions: Individual solutions of 13, 16, 18, 21, and 23 mg/mL of sucrose from USP Sucrose RS, in water
Sample solution: Transfer about 1.875 g of Injection to a 25-mL ask. Add 1.25 mL of water and mix. Add 1.25 mL of a monobasic
sodium phosphate solution, prepared by dissolving 30 g in 50 mL, and mix. Allow the resulting solution to stand for 10 min to
precipitate the colloidal ferric hydroxide. Dilute with water to volume. Centrifuge this solution at 3000 rpm for 15 min. Pass the
resulting solution through a lter, discarding the rst 2 mL of the ltrate.
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC
Detector: Refractive index
Column: 4-mm × 25-cm; packing L8
Temperatures
Detector: 20–25° (±2°)
Column: 20–25° (±2°)
Flow rate: 2 mL/min
Injection volume: 20 µL
System suitability
Samples: Standard solutions
[ NOTE— The retention time for sucrose is about 8 min.]
Suitability requirements
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Correlation coe cient: NLT 0.998 from the linear regression of the Standard solutions
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Analysis
Samples: Standard solutions and Sample solution
Plot the peak area for each Standard solution versus concentration of sucrose in mg/mL, and draw the straight line best tting the
ve plotted points. From the graph, determine the concentration of sucrose, in mg/mL, in the Sample solution.
Calculate the quantity of sucrose, in mg, in each mL of Injection taken:
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Result = (CU × D × G)/W

CU = concentration of sucrose in the Sample solution (mg/mL)

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D = dilution volume of the Sample solution (mL)

G = density of Injection taken (g/mL)

W = weight of Injection taken (g)

Acceptance criteria: 260–340 mg/mL

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• IRON
Solution A: Transfer 2.64 g of calcium chloride to a 1000-mL volumetric ask, add 500 mL of water, and swirl to dissolve. Add 5.0 mL
of hydrochloric acid, and dilute with water to volume.
Standard stock solution: 50 µg/mL of iron prepared as follows. Transfer about 350 mg of ferrous ammonium sulfate to a 1000-mL
volumetric ask. Add water to dissolve, dilute with water to volume, and mix.
Standard solutions: Individual solutions containing 2.0, 4.0, 6.0, 8.0, and 10.0 µg/mL of iron in Solution A from the Standard stock
solution
Sample stock solution: Using a “to contain” pipet, transfer 2.0 mL of Injection to a 100-mL volumetric ask. Rinse the pipet several
times with Solution A. Add 5 mL of hydrochloric acid, and swirl until the solution turns yellow. After the solution has cooled to room
temperature, dilute with Solution A to volume, and mix.
Sample solution: Nominally 8.0 µg/mL of iron prepared as follows. Pipet 2.0 mL of the Sample stock solution to a 100-mL volumetric
ask, and dilute with Solution A to volume.
Instrumental conditions
(See Atomic Absorption Spectroscopy 〈852〉.)
Mode: Atomic absorption spectrophotometry
Analytical wavelength: 248.3 nm iron emission line
Lamp: Iron hollow-cathode
Flame: Air–acetylene
Blank: Solution A

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Analysis
Samples: Standard solutions and Sample solution
Plot the absorbance of each Standard solution versus concentration, in µg/mL, of iron, and draw the straight line best tting the
ve plotted points. From the graph, determine the concentration, in µg/mL, of iron in the Sample solution.
Calculate the percentage of the labeled amount of iron in each mL of Injection taken:

Result = (CA/CU) × 100

CA = actual concentration of iron in the Sample solution determined from the calibration curve (µg/mL)

CU = nominal concentration of iron in the Sample solution (µg/mL)

Acceptance criteria: 95.0%–105.0%

OTHER COMPONENTS
• CONTENT OF CHLORIDE
Sample: About 12 g of Injection
Analysis: Transfer the Sample to a 50-mL beaker. Add 40 mL of water and 0.3 mL of 65% nitric acid, and, while stirring, titrate with 0.01
N silver nitrate VS, determining the endpoint potentiometrically with silver-glass electrodes.
Calculate the content of chloride, in mg, of Injection taken. Each mL of 0.01 N silver nitrate consumed is equal to 0.3545 mg of
chloride (Cl).
Acceptance criteria: 0.012%–0.025%

IMPURITIES
• LIMIT OF IRON [FE(II)]
Supplementary electrolyte solution: Dissolve 15.0 g of sodium acetate in 100 mL of water and adjust with 0.1 N acetic acid to a pH of
7.0.
Sample solution: Volume of Injection equivalent to 20–120 µg/mL of elemental iron
Analysis: Transfer a suitable amount of Supplementary electrolyte solution to a polarographic cell equipped with a mercury drop

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electrode. With the electrode submerged in the liquid, bubble nitrogen through the liquid for 5 min. Avoiding any undue exposure to
air, immediately transfer the Sample solution to the polarographic cell. The sample must be analyzed immediately upon opening the
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container.
Record the polarogram from 0 mV and −1700 mV. The iron [Fe(III)/Fe(II)] peak is detected at −750 ± 50 mV and the iron [Fe(II)/Fe(0)]
peak is detected at −1400 ± 50 mV. Measure the iron [Fe(II)/Fe(III)] peak responses obtained from the polarogram, and perform a
blank determination.
Calculate the content of iron [Fe(II)], in % w/v, in the volume of Injection taken:
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Result = [1 − (2/R)] × CT

R = peak response ratio of iron [Fe(II)] to iron [Fe(III)]

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CT = total iron concentration of the Injection (% w/v)

Acceptance criteria: NMT 0.4%

SPECIFIC TESTS
• PH 〈791〉: 10.5–11.1 at 20°
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• TURBIDITY
Sample solution: Transfer 0.5 g of Injection to a 150-mL beaker. Add 100 mL of water and, with constant stirring, adjust with 0.1 N
hydrochloric acid VS to a pH of 6.0.
Analysis: Remove the pH electrode from the solution. Adjust a light source such that the beam hits the beaker at a parallel angle 2 cm
below the surface of the liquid. The light must shine through to the surface, and the solution must not have any turbidity.
Measurement must be carried out in a room as dark as possible. Slowly add 0.1 N hydrochloric acid VS, dropwise, until a slight but
lasting turbidity develops. Record the pH of the solution as the turbidity point of the Injection.
Acceptance criteria: 4.4–5.3
• ABSENCE OF LOW-MOLECULAR WEIGHT IRON [FE(II) AND FE(III)] COMPLEXES: In the polarograms obtained in the test for Limit of Iron [Fe(II)], no
additional peaks are found.
• ALKALINITY
Sample solution: 5 mL of Injection
Analysis: Titrate the Sample solution with 0.1 N hydrochloric acid VS with constant stirring to a pH of 7.4. Record the volume of 0.1 N
hydrochloric acid VS consumed, and calculate the alkalinity of the Injection as the volume of acid, in mL, consumed per mL of
Injection.
Acceptance criteria: 0.5–0.8 mL of 0.1 N hydrochloric acid VS is consumed per mL of Injection.
• OSMOLALITY AND OSMOLARITY 〈785〉, Osmolarity
Sample solution: Dilute Injection in water (1 in 10).
Acceptance criteria: 1150–1350 mOsmol/L

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11/2/2020 USP-NF Iron Sucrose Injection

• SPECIFIC GRAVITY 〈841〉: 1.135–1.165 at 20°

• PARTICULATE MATTER IN INJECTIONS 〈788〉, Method 1 Light Obscuration Particle Count Test
Sample solution: Prepare a solution of Injection (1 in 40) using water that has been passed through a lter having a 1.2-µm or ner
pore size.
Acceptance criteria: Meets the requirements for small-volume injections
• BACTERIAL ENDOTOXINS TEST 〈85〉: NMT 3.7 USP Endotoxin Units/mg of iron contained in Injection
• OTHER REQUIREMENTS: Meets the requirements in Injections and Implanted Drug Products 〈1〉

ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in single-dose containers of Type I glass. Store at controlled room temperature. Do not freeze.
• LABELING: Label it to indicate that it is for intravenous use only, and that when administered by intravenous infusion, the Injection must
be diluted with 0.9% Sodium Chloride Injection to a concentration of 1.0–2.0 mg/mL of elemental iron. Label it also to state the total
osmolarity of the solution expressed in mOsmol/L.
• USP REFERENCE STANDARDS 〈11〉
USP Sucrose RS

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

Topic/Question Contact Expert Committee

IRON SUCROSE INJECTION Tsion Bililign SM22020 Small Molecules 2

Scienti c Liaison

Chromatographic Database Information: Chromatographic Database

Most Recently Appeared In:

Pharmacopeial Forum: Volume No. 41(6)
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Page Information:

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