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Printed by: Nguyen Nhan Official Date: Official as of 01-Jul-2020 Document Type: USP @2021 USPC
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Sensitivity solution: 0.25 µg/mL of USP Atropine Sulfate RS

Atropine Sulfate in Diluent
Standard solution: 0.5 mg/mL of USP Atropine Sulfate RS
Change to read: in Diluent
Sample solution: 0.5 mg/mL of Atropine Sulfate in Diluent
Chromatographic system
(See Chromatography á621ñ, System Suitability.)
Mode: LC
Detector: UV 210 nm
Column: 4.6-mm × 15-cm; 3-µm packing L1
Column temperature: 50°
Flow rate: 2 mL/min
Injection volume: 5 µL
System suitability
Samples: System suitability solution, Sensitivity solution, and
Standard solution
(C17H23NO3)2 · H2SO4 · H2O ▲
694.84▲ (ERR 1-Jul-2020) [NOTE—See Table 2 for the relative retention times.]
Anhydrous 676.82 Suitability requirements
Benzeneacetic acid, α-(hydroxymethyl)-, 8-methyl-8- Resolution: NLT 1.4 between hyoscyamine related
azabicyclo[3.2.1]oct-3-yl ester, endo-(±)-, sulfate (2:1) (salt), compound A and atropine, System suitability solution
monohydrate; Tailing factor: 0.8–1.8 for atropine, System suitability
1αH,5αH-Tropan-3-α-ol (±)-tropate (ester), sulfate (2:1) (salt) solution

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monohydrate [5908-99-6]. Relative standard deviation: NMT 0.73%, Standard
Anhydrous [55-48-1]. solution
Signal-to-noise ratio: NLT 10 for atropine, Sensitivity
DEFINITION solution
Atropine Sulfate contains NLT 98.0% and NMT 102.0% of Analysis
atropine sulfate [(C17H23NO3)2 · H2SO4], calculated on the Samples: Standard solution and Sample solution
anhydrous basis.
[CAUTION—Handle atropine sulfate with exceptional care,
because it is highly potent.]
ci Calculate the percentage of atropine sulfate [(C17H23NO3)2
· H2SO4] in the portion of Atropine Sulfate taken:

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

• A. SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared
ffi
Spectroscopy: 197K or 197A rU = peak response of atropine from the Sample
• B. IDENTIFICATION TESTS—GENERAL á191ñ, Chemical solution
Identification Tests, Sulfate rS = peak response of atropine from the Standard
Sample solution: 50 mg/mL solution
Acceptance criteria: Meets the requirements CS = concentration of USP Atropine Sulfate RS in the
• C. The retention time of the major peak of the Sample Standard solution (mg/mL)
O

solution corresponds to that of the System suitability CU = concentration of Atropine Sulfate in the Sample
solution, as obtained in the Assay. solution (mg/mL)

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

• PROCEDURE basis
Buffer: 1.8 g/L of monobasic potassium phosphate and
IMPURITIES
2.5 g/L of sodium 1-pentanesulfonate, adjusted with
• RESIDUE ON IGNITION á281ñ: NMT 0.2%
phosphoric acid to a pH of 2.5
• ORGANIC IMPURITIES
Diluent: Acetonitrile and Buffer (20:80)
Buffer, Diluent, Solution A, Solution B, Mobile phase,
Solution A: Acetonitrile and Buffer (5:95)
System suitability solution, Sensitivity solution,
Solution B: Acetonitrile and Buffer (80:20)
Standard solution, Sample solution, Chromatographic
Mobile phase: See Table 1.
system, and System suitability: Proceed as directed in the
Table 1
Assay.
Analysis
Time Solution A Solution B Sample: Sample solution
(min) (%) (%)
Calculate the percentage of each impurity in the portion of
0 92 8 Atropine Sulfate taken:
11 79 21
Result = (rU/rT) × (1/F) × 100
15 46 54
rU = peak response of each impurity from the Sample
15.1 92 8
solution
20 92 8 rT = sum of all the peak responses from the Sample
solution
[NOTE—The gradient was established on an HPLC system F = relative response factor for each impurity (see
with a dwell volume of approximately 0.8 mL.] Table 2)
System suitability solution: 1 µg/mL of USP Hyoscyamine
Related Compound A RS and 0.5 mg/mL of USP Atropine Acceptance criteria: See Table 2. The reporting threshold is
Sulfate RS in Diluent 0.05%.

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2

Table 2 d (1R,3S,5R)-6-Hydroxy-8-methyl-8-azabicyclo[3.2.1]octan-3-yl (S)-3-hydroxy-

2-phenylpropanoate; also known as (1R,3S,5R,6RS)-6-Hydroxy-8-methyl-8-
Relative Relative Acceptance azabicyclo[3.2.1]oct-3-yl (2S)-3-hydroxy-2-phenylpropanoate.
Retention Response Criteria, e (1R,3r,5S)-8-Methyl-8-azabicyclo[3.2.1]octan-3-yl 2-hydroxy-3-
Name Time Factor NMT (%)
phenylpropanoate; also known as (1R,3r,5S)-8-Methyl-8-azabicyclo[3.2.1]oct-
Tropic acida 0.56 2.1 0.2 3-yl (2RS)-2-hydroxy-3-phenylpropanoate.
f (1R,3r,5S)-8-Methyl-8-azabicyclo[3.2.1]octan-3-yl 2-phenylacrylate; also
7-Hydroxyhyoscyamineb 0.66 1.0 0.2 known as (1R,3r,5S)-8-Methyl-8-azabicyclo[3.2.1]oct-3-yl 2-phenylpropenoate.
Scopolaminec 0.72 1.0 0.2
SPECIFIC TESTS
6-Hydroxyhyoscyamined 0.75 1.0 0.2 • OPTICAL ROTATION á781Sñ, Procedures, Specific Rotation
Hyoscyamine related com-
Sample solution: 0.1 g/mL of Atropine Sulfate in water
pound A 0.97 1.2 0.3 Temperature: 20°
Path length: 1.0 or 2.0 dm
Atropine 1.0 1.0 — Acceptance criteria: Between −0.50° and +0.05°
Littorine e
1.13 1.2 0.2 • WATER DETERMINATION á921ñ, Method I: NMT 4.0%
Apoatropinef 1.60 2.0 0.2 ADDITIONAL REQUIREMENTS
• PACKAGING AND STORAGE: Preserve in tight, light-resistant
Any individual,
unspecified impurity
—
1.0 0.1 containers.
• USP REFERENCE STANDARDS á11ñ
Total impurities — — 0.5 USP Atropine Sulfate RS
a 3-Hydroxy-2-phenylpropanoic acid; also known as (2RS)-3-Hydroxy-2-
USP Hyoscyamine Related Compound A RS
Norhyoscyamine sulfate; (1R,3r,5S)-8-Azabicyclo[3.2.1]

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phenylpropanoic acid.
b (1S,3R,5S)-6-Hydroxy-8-methyl-8-azabicyclo[3.2.1]oct-3-yl (S)-3-hydroxy-2- octan-3-yl (S)-3-hydroxy-2-phenylpropanoate sulfate
phenylpropanoate; also known as (1S,3R,5S,6RS)-6-Hydroxy-8-methyl-8- (2:1).
azabicyclo[3.2.1]oct-3-yl (2S)-3-hydroxy-2-phenylpropanoate. (C16H21NO3)2 · H2SO4 648.77
c (S)-(1R,2R,4S,5S,7s)-9-Methyl-3-oxa-9-azatricyclo[3.3.1.02,4]nonan-7-yl 3-
hydroxy-2-phenylpropanoate; also known as (S)-(1R,2R,4S,5S,7s)-9-Methyl-3-
oxa-9-azatricyclo[3.3.1.02,4]non-7-yl (2S)-3-hydroxy-2-phenylpropanoate.
ci
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