First Supplement to USP 40–NF 35 Official Monographs / Timolol 1

. Mode: LC
Timolol Maleate Detector: UV 295 nm
Column: 2.1-mm × 10-cm; 2.6-µm packing L1
Autosampler temperature: 4°
Flow rate: 0.4 mL/min
Injection volume: 2.5 µL
System suitability
Samples: System suitability solution and Standard
solution
Suitability requirements
C13H24N4O3S · C4H4O4 432.49 Resolution: NLT 2 between timolol and timolol re-
2-Propanol, 1-[(1,1-dimethylethyl)amino]-3- lated compound D, System suitability solution
[[4-(4-morpholinyl)-1,2,5-thiadiazol-3-yl]oxy]-, (S)-, (Z)- Tailing factor: NMT 2.0, Standard solution
2-butenedioate (1:1) (salt); Relative standard deviation: NMT 0.73%, Standard
(−)-1-(tert-Butylamino)-3-[(4-morpholino-1,2,5-thiadiazol- solution
3-yl)oxy]-2-propanol maleate (1:1) (salt) [26921-17-5]. Analysis
Samples: Standard solution and Sample solution
DEFINITION Calculate the percentage of timolol maleate
Timolol Maleate contains NLT 98.0% and NMT 102.0% of (C13H24N4O3S · C4H4O4) in the portion of Timolol
timolol maleate (C13H24N4O3S · C4H4O4), calculated on the Maleate taken:
dried basis.
Result = (rU/rS) × (CS/CU) × 100
IDENTIFICATION
• A. INFRARED ABSORPTION 〈197M〉 rU = peak response of timolol from the Sample
solution
rS = peak response of timolol from the Standard
Delete the following: solution
CS = concentration of USP Timolol Maleate RS in
■ • B. ULTRAVIOLET ABSORPTION 〈197U〉
.

the Standard solution (µg/mL)

Analytical wavelength: 294 nm CU = concentration of Timolol Maleate in the
Sample solution: 25 µg/mL in 0.12 N hydrochloric acid Sample solution (µg/mL)
Acceptance criteria: Absorptivities, calculated on the Acceptance criteria: 98.0%–102.0% on the dried basis
dried basis, do not differ by more than 3.0%.■1S (USP40)
IMPURITIES
Change to read: • RESIDUE ON IGNITION 〈281〉: NMT 0.1%

• ■B.■1S (USP40) The retention time of the major peak of the

. Delete the following:
Sample solution corresponds to that of the Standard solu-
tion, as obtained in the ■Enantiomeric Purity test.■1S (USP40)
.
•• HEAVY METALS 〈231〉, Method II: 20 ppm• (Official 1-Jan-2018)
.

• ORGANIC IMPURITIES
ASSAY Solution A, Solution B, Mobile phase, Diluent, and
• PROCEDURE Chromatographic system: Proceed as directed in the
Solution A: Dilute 0.5 mL of trifluoroacetic acid with Assay.
water to 1 L. System suitability solution: 100 µg/mL each of USP
Solution B: Dilute 0.5 mL of trifluoroacetic acid with Timolol Maleate RS, USP Timolol Related Compound B
acetonitrile to 1 L. RS, USP Timolol Related Compound C RS, USP Timolol
Mobile phase: See Table 1. Related Compound D RS, USP Timolol Related Com-
pound E RS, and USP Timolol Related Compound F RS
Table 1 in Diluent. [NOTE—Prepare fresh and analyze immedi-
Time Solution A Solution B
ately as USP Timolol Related Compound E RS degrades
(min) (%) (%)
rapidly.]
Standard solution: 1 µg/mL of USP Timolol Maleate RS
0 84 16 and 4 µg/mL each of USP Timolol Related Compound B
2.4 84 16 RS, USP Timolol Related Compound C RS, USP Timolol
8 20 80 Related Compound D RS, USP Timolol Related Com-
8.1 84 16 pound E RS, and USP Timolol Related Compound F RS
11 84 16 in Diluent. Sonicate if needed for 0.5 min.
Sample solution: 1 mg/mL of Timolol Maleate in
Diluent: Methanol and water (60:40) Diluent
System suitability solution: 100 µg/mL of USP Timolol System suitability
Maleate RS and 10 µg/mL of USP Timolol Related Com- Samples: System suitability solution and Standard
pound D RS in Diluent solution
Standard solution: 100 µg/mL of USP Timolol Maleate Suitability requirements
RS in Diluent Resolution: NLT 2.0 between timolol and timolol re-
Sample solution: 100 µg/mL of Timolol Maleate in lated compound D, System suitability solution
Diluent Relative standard deviation: NMT 4.0% for timolol,
Chromatographic system Standard solution
(See Chromatography 〈621〉, System Suitability.) Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each specified impurity in
the portion of Timolol Maleate taken:
Result = (rU/rS) × (CS/CU) × 100

© 2017 The United States Pharmacopeial Convention All Rights Reserved.

2 Timolol / Official Monographs First Supplement to USP 40–NF 35

rU = peak response of each specified impurity from Signal-to-noise ratio: NLT 10 for timolol related
the Sample solution compound A
rS = peak response of the corresponding USP Analysis
Reference Standard from the Standard Samples: Standard solution and Sample solution
solution Calculate the percentage of timolol related compound
CS = concentration of the corresponding USP A in the portion of Timolol Maleate taken:
Reference Standard in the Standard solution
(µg/mL) Result = (rU/rS) × (CS/CU) × 100
CU = concentration of Timolol Maleate in the
Sample solution (µg/mL) rU = peak response of timolol related compound A
Calculate the percentage of any individual unspecified from the Sample solution
impurity in the portion of Timolol Maleate taken: rS = peak response of timolol related compound A
from the Standard solution
Result = (rU/rS) × (CS/CU) × 100 CS = concentration of USP Timolol Related
Compound A RS in the Standard solution
rU = peak response of any individual unspecified (mg/mL)
impurity from the Sample solution CU = concentration of Timolol Maleate in the
rS = peak response of timolol from the Standard Sample solution (mg/mL)
solution Acceptance criteria: NMT 1.0%■1S (USP40)
CS = concentration of USP Timolol Maleate RS in
the Standard solution (µg/mL) SPECIFIC TESTS
CU = concentration of Timolol Maleate in the
Sample solution (µg/mL) Delete the following:
Acceptance criteria: See Table 2. Disregard any impu-
rity peaks less than 0.05%. ■• OPTICAL ROTATION 〈781S〉, Specific Rotation
.

Sample solution: 50 mg/mL of Timolol Maleate in 1.0

Table 2 N hydrochloric acid
Relative Acceptance Acceptance criteria: −11.7° to −12.5° (λ = 405
Retention Criteria, nm)■1S (USP40)
Name Time NMT (%) • PH 〈791〉
Sample solution: 20 mg/mL of Timolol Maleate in
Timolol related compound B 0.5 0.4
water
Timolol related compound D 0.8 0.4 Acceptance criteria: 3.8–4.3
Timolol maleate 1.0 — • LOSS ON DRYING 〈731〉
Timolol related compound E 1.4 0.4 Analysis: Dry under vacuum at 100° to constant
Timolol related compound C 1.8 0.4 weight.
Timolol related compound F 2.0 0.4 Acceptance criteria: NMT 0.5%
Any unspecified impurity — 0.10 ADDITIONAL REQUIREMENTS
Total degradation products — 1.0 • PACKAGING AND STORAGE: Preserve in well-closed
containers.

Add the following: Change to read:

■ • ENANTIOMERIC PURITY
.

• USP REFERENCE STANDARDS 〈11〉

Prepare all solutions in low-actinic glassware protected USP Timolol Maleate RS
from light. ■USP Timolol Related Compound A RS
Mobile phase: Diethylamine, 2-propanol, and hexane
.

(R)-1-(tert-Butylamino)-3-(4-morpholino-1,2,5-thiadiazol-
(2:40:960) 3-yloxy)propan-2-ol.
Diluent: Methylene chloride and 2-propanol (25:75) C13H24N4O3S 316.42■1S (USP40)
System suitability solution: 0.03 mg/mL each of USP USP Timolol Related Compound B RS
Timolol Maleate RS and USP Timolol Related Com- 3-(tert-Butylamino)-2-(4-morpholino-1,2,5-thiadiazol-
pound A RS in Diluent 3-yloxy)propan-1-ol.
Standard solution: 0.03 mg/mL of USP Timolol Related C13H24N4O3S 316.42
Compound A RS in Diluent USP Timolol Related Compound C RS
Sample solution: 3 mg/mL of Timolol Maleate in N-(tert-Butyl)-2,3-bis(4-morpholino-1,2,5-thiadiazol-
Diluent 3-yloxy)propan-1-amine.
Chromatographic system C19H31N7O4S2 485.19
(See Chromatography 〈621〉, System Suitability.) USP Timolol Related Compound D RS
Mode: LC 4-Morpholino-1,2,5-thiadiazol-3-ol.
Detector: UV 297 nm C6H9N7O4S 187.22
Column: 4.6-mm × 25-cm; 5-µm packing L40 USP Timolol Related Compound E RS
Autosampler temperature: 4° (S)-3-(tert-Butylamino)-1-(4-morpholino-1,2,5-thiadiazol-
Flow rate: 1 mL/min 3-yloxy)propan-2-yl hydrogen maleate.
Injection volume: 5 µL C17H26N4O6S 414.48
System suitability USP Timolol Related Compound F RS
Sample: System suitability solution 3-Chloro-4-morpholino-1,2,5-thiadiazol.
Suitability requirements C6H8ClN3OS 205.67
Resolution: NLT 4.0 between the timolol related
compound A and timolol peaks
Relative standard deviation: NMT 1.5% for both
peaks

© 2017 The United States Pharmacopeial Convention All Rights Reserved.