Amikacin Sulfate

(a'' mi kay' sin sul' fate)

C22H43N5O13·H2SO4

762.15

C22H43N5O13·2H2SO4

781.76
d-Streptamine, O-3-amino-3-deoxy- -d-glucopyranosyl-(1®6)-O-[6-amino-6-deoxy- -d-
glucopyranosyl-(1®4)]-N1-(4-amino-2-hydroxy-1-oxobutyl)-2-deoxy-, (S)-, sulfate (1:2 or 1:1.8)
(salt);    
O-3-Amino-3-deoxy- -d-glucopyranosyl-(1®4)-O-[6-amino-6-deoxy- -d-glucopyranosyl-
(1®6)]-N3-(4-amino-l-2-hydroxybutyryl)-2-deoxy-l-streptamine sulfate (1:2 or 1:1.8))    
[39831-55-5].

DEFINITION

Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:2 contains the equivalent of
NLT 674 µg and NMT 786 µg of amikacin (C22H43N5O13) per mg, calculated on the dried
basis; Amikacin Sulfate having a molar ratio of amikacin to H2SO4 of 1:1.8 contains the
equivalent of NLT 691 µg and NMT 806 µg of amikacin (C22H43N5O13) per mg, calculated on
the dried basis.

IDENTIFICATION
•  A. Thin-Layer Chromatographic Identification Test 201

Standard solution and Sample solution:  6 mg/mL in water

Solution A:  Standard solution and Sample solution (1:1)

Application volume:  3 µL

Developing solvent system:  Methanol, chloroform, and ammonium hydroxide (12:5:7)

Spray reagent:  10 mg/mL of ninhydrin in a mixture of butyl alcohol and pyridine (100:1)

Analysis 

Samples:  Standard solution, Sample solution, and Solution A

Proceed as directed in the chapter, except to develop the chromatogram by continuous flow
for 5.5 h. Remove the plate from the chamber, allow the solvent to evaporate, and heat the
plate at 110 for 15 min. Spray the plate with Spray reagent, and immediately locate the spots.

Acceptance criteria:  Amikacin appears as a pink spot, and the spots of the Sample solution and
Solution A correspond in distance from the origin to that of the Standard solution.

•  B. The retention time of the peak for amikacin of the Sample solution corresponds to that of
the Standard solution, as obtained in the Assay.

ASSAY

•  Procedure

Mobile phase:  0.115 N sodium hydroxide

System suitability solution:  0.02 mg/mL of USP Amikacin RS and 0.008 mg/mL of USP
Kanamycin Sulfate RS in water

Standard solution:  0.02 mg/mL of USP Amikacin RS in water

Sample solution:  Equivalent to 0.02 mg/mL of amikacin, from Amikacin Sulfate, in water

Chromatographic system 

(See Chromatography 621 , System Suitability.)

Mode:  LC

Detector:  Electrochemical detector, a gold working electrode, and a pH silver–silver chloride

reference electrode. [Note—The electrochemical detector is used in the integrated
amperometric mode with a range of 300 nC, an output of 1 V full scale, a rise time of 0.5 s,
positive polarity, potential E = 0.04 V; t1 = 200 ms; E2 = 0.8 V; t2 = 190 ms; E3 = 0.8 V; and t3
= 190 ms. ]

Column 

Guard column:  Packing L47

Analytical column:  4-mm × 25-cm; packing L47

Flow rate:  0.5 mL/min

Injection size:  20 µL

System suitability 

Samples:  System suitability solution and Standard solution

[Note—The relative retention times for kanamycin and amikacin are 0.8 and 1.0, respectively. ]

Suitability requirements 

Resolution:  NLT 3 between kanamycin and amikacin, System suitability solution

Tailing factor:  NMT 2, Standard solution

Relative standard deviation:  NMT 3%, Standard solution

Analysis 

Samples:  Standard solution and Sample solution

Calculate the quantity, in µg, of C22H43N5O13 in each mg of Amikacin Sulfate taken:

Result = (rU/rS) × (CS/CU) × E

rU = = peak area from the Sample solution

rS = = peak area from the Standard solution

CS = = concentration of USP Amikacin RS in the Standard

solution (mg/mL)

CU = = concentration for the Sample solution (mg/mL)

E = = designated amikacin content of USP Amikacin RS

(µg/mg)

Acceptance criteria: 

Ratio of Acceptance Criteria

amikacin/H2SO4 (µg (C22H43N5O13)/mg) on the dried basis

1:2 674–786

1:1.8 691–806

IMPURITIES

Inorganic Impurities 

•  Residue on Ignition 281 : NMT 1.0%, the charred residue being moistened with 2 mL of
nitric acid and 5 drops of sulfuric acid

SPECIFIC TESTS

•  Optical Rotation, Specific Rotation 781 : +76 to +84

Sample solution:  20 mg/mL, in water

•  Crystallinity 695 : Meets the requirements

•  pH 791 : 2.0–4.0 (1:2 salt), or 6.0–7.3 (1:1.8 salt), in a solution containing 10 mg/mL

•  Loss on Drying 731 : Dry 100 mg, in a vacuum at a pressure not exceeding 5 mm of
mercury at 110 for 3 h: it loses NMT 13.0% of its weight.

ADDITIONAL REQUIREMENTS

•  Packaging and Storage: Preserve in tight containers.

•  Labeling: Label it to indicate whether its molar ratio of amikacin to H2SO4 is 1:2 or 1:1.8.

•  USP Reference Standards 11

USP Amikacin RS

USP Kanamycin Sulfate RS

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

Topic/Question Contact Expert Committee

Monograph Ahalya Wise, M.S. (MDANT05) Monograph Development-

Scientist Antibiotics
1-301-816-8161

Reference Lili Wang, Technical Services

Standards Scientist
1-301-816-8129
RSTech@usp.org

USP33–NF28 Page 1939

Chromatographic Column— 

AMIKACIN SULFATE

Chromatographic columns text is not derived from, and not part of, USP 33 or NF 28.