Accessed from 10.6.1.

1 by JNJconsumer on Wed Jun 28 18:24:31 EDT 2017

NF 35 Official Monographs / Benzalkonium 7533

• USP REFERENCE STANDARDS 〈11〉 Flow rate: 2 mL/min

USP Benzaldehyde RS Injection volume: 20 µL
USP Benzalkonium Chloride RS System suitability
USP Benzyl Alcohol RS Sample: Standard solution
[NOTE—See Table 1. Relative retention times are pro-
vided for information only, and the Standard should
be used to ensure appropriate peak identification.]
.

Benzalkonium Chloride Solution Table 1

Relative
DEFINITION Retention
Benzalkonium Chloride Solution contains NLT 95.0% and Name Time
NMT 105.0% of the labeled amount of benzalkonium
chloride in a solution having a concentration of 1.0% or C10 homolog 0.9
more; and NLT 93.0% and NMT 107.0% of the labeled C12 homolog 1.0
amount of benzalkonium chloride in a solution having a C14 homolog 1.3
concentration of less than 1.0%. It may contain a suitable C16 homolog 1.7
coloring agent and may contain NMT 10% of alcohol.
[CAUTION—Mixing Benzalkonium Chloride Solution with or- Suitability requirements
dinary soaps and anionic detergents may decrease or de- Resolution: NLT 1.5 between the C12 and C14
stroy the bacteriostatic activity of the Solution.] homologs
Relative standard deviation: NMT 2.0% for the C12
IDENTIFICATION homolog
• A. Analysis
Analysis: To 2 mL of a solution having an equivalent of Samples: Standard solution and Sample solution
10 mg/mL of benzalkonium chloride add 1 mL of 2 N Identify the homolog peaks by comparison of the reten-
nitric acid. tion times from the Sample solution with those of the
Acceptance criteria: A white precipitate is formed and Standard solution.
is dissolved after adding 5 mL of alcohol. Calculate the percentage of each quaternary ammo-
• B. IDENTIFICATION TESTS—GENERAL 〈191〉, Chloride: A solu- nium homolog in the portion of Solution taken:
tion of it in a mixture of equal volumes of water and
alcohol meets the requirements.
• C.
Analysis: Dissolve the residue, obtained by evaporating
on a steam bath a volume of Solution equivalent to
200 mg of benzalkonium chloride, in 1 mL of sulfuric rU = peak area of each homolog from the Sample
acid. Add 100 mg of sodium nitrate, and heat on a solution
steam bath for 5 min. Cool, dilute with water to 10 mL, Mr = molecular weight of each homolog. The
add 500 mg of zinc dust, and warm for 5 min on a molecular weights of the C10, C12, C14, and
steam bath. To 2 mL of the clear supernatant add 1 mL C16 homologs are 312, 340, 368, and 396,
of sodium nitrite solution (1 in 20), cool in ice water, respectively.
then add 3 mL of a solution of 500 mg of 2-naphthol in Acceptance criteria: On the solid basis, the content of
10 mL of 6 N ammonium hydroxide. the n-C12H25 homolog is NLT 40.0%, and the content of
Acceptance criteria: An orange-red color is produced. the n-C14H29 homolog is NLT 20.0% of the total alkyl-
• D. CHROMATOGRAPHIC IDENTITY benzyldimethylammonium chloride content. The
Analysis: Proceed as directed in the test for Ratio of amount of the n-C12H25 and n-C14H29 homolog compo-
Alkyl Components. nents together is NLT 70.0% of the total alkylbenzyldi-
Acceptance criteria: The retention times of the major methylammonium chloride content.
peaks for benzalkonium chloride of the Sample solution • TOTAL ALKYLBENZYLDIMETHYLAMMONIUM CHLORIDES
correspond to those of the Standard solution. Sample solution: Evaporate or dilute with water to
30 mL a volume of Solution equivalent to 500 mg of

NF Monographs
ASSAY
• RATIO OF ALKYL COMPONENTS benzalkonium chloride.
Solution A: Adjust a 0.1 M solution of sodium acetate Analysis: Transfer the Sample solution, with the aid of a
with glacial acetic acid to a pH of 5.0. minimum quantity of water, to a glass-stoppered,
Mobile phase: Acetonitrile and Solution A (9:11). Aceto- 250-mL conical separator. Transfer 25 mL of methylene
nitrile and Solution A may be adjusted from (2:3) to chloride. Add 10 mL of 0.1 N sodium hydroxide, and
(3:2) to meet system suitability requirements. 10.0 mL of freshly prepared potassium iodide solution
Standard solution: 4 mg/mL of benzalkonium chloride (1 in 20), insert the stopper in the separator, shake,
prepared from USP Benzalkonium Chloride RS and allow the layers to separate, and discard the methylene
water chloride layer. Wash the aqueous layer with three
Sample solution: Transfer a volume of Solution, equiva- 10-mL portions of methylene chloride, and discard the
lent to 400 mg of benzalkonium chloride, to a 100-mL washings. Transfer the aqueous layer to a glass-stop-
volumetric flask, and dilute with water to volume. pered, 250-mL conical flask, and rinse the separator
Chromatographic system with three 5-mL portions of water, adding the washings
(See Chromatography 〈621〉, System Suitability.) to the flask. Add 40 mL of cold hydrochloric acid to the
Mode: LC flask, mix, and titrate with 0.05 M potassium iodate VS
Detector: UV 254 nm until the solution becomes light brown in color. Add
Column: 3.9-mm × 30-cm; packing L10 or 4.6-mm × 5 mL of methylene chloride, insert the stopper into the
25-cm; 10-µm packing L10 flask, and shake vigorously. Continue the titration,
dropwise, with shaking after each addition, until the
methylene chloride layer no longer changes color and
the aqueous layer is clear yellow. Record the titrant vol-
ume, VT, in mL. Perform a blank determination, using
20 mL of water as the sample, and record the titrant

Official from May 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.
Accessed from 10.6.1.1 by JNJconsumer on Wed Jun 28 18:24:31 EDT 2017

7534 Benzalkonium / Official Monographs NF 35

volume, VB, in mL. [NOTE—VB > VT.] The difference be- Table 3
tween the two titrations represents the amount of po- Hold Time at
tassium iodate equivalent to the weight of benzalko- Initial Flow Flow Ramp Final Flow Final Flow
nium chloride in the sample. Each mL of 0.05 M (mL/min) (mL/min2) (mL/min) (min)
potassium iodate is equivalent to x/10 mg of benzalko-
.

nium chloride, where x represents the average molecu- 1.4 — 1.4 6

lar weight of the sample, derived by summing, for all 1.4 5 3 8.6
homologs, the products:
Injection volume: 0.5 µL
Injection type: Split, 75:1
Inlet liner: 4-mm liner with deactivated glass wool
System suitability
rU = peak area of each homolog from the test for Sample: Standard solution containing 0.0012 g/mL of
Ratio of Alkyl Components alcohol (C2H5OH)
rT = sum of all the peak areas of the homologs [NOTE—The relative retention times for alcohol,
from the test for Ratio of Alkyl Components 2-propanol, and tertiary butyl alcohol are 0.75, 0.90,
Mr = molecular weight of each homolog. The and 1.0, respectively.]
molecular weights of the C10, C12, C14, and Suitability requirements
C16 homologs are 312, 340, 368, and 396, Resolution: NLT 2.0 between 2-propanol and tertiary
respectively. butyl alcohol
Acceptance criteria Relative standard deviation: NMT 2% for the peak
For labeled concentrations of NLT 1.0%: response ratio of alcohol to the internal standard
95.0%–105.0% Analysis
For labeled concentrations less than 1.0%: Samples: Standard solutions and Sample solution
93.0%–107.0% Plot the peak response ratios of alcohol to tertiary butyl
alcohol in the Standard solutions versus the content, in
OTHER COMPONENTS g/mL, of alcohol, and draw the straight line best fit-
• ALCOHOL CONTENT (if added) ting the plotted points. From the graph obtained, de-
Diluent: 2-Propanol and water (8:2) termine the content, C, in g/mL, of alcohol (C2H5OH)
Internal standard solution: 0.005 g/mL of tertiary bu- in the Sample solution.
tyl alcohol in water Calculate the percentage of alcohol (C2H5OH) in the
Alcohol stock solution: 0.015 g/mL of alcohol portion of Solution (v/v) taken:
(C2H5OH) in water
Standard solutions: Introduce 1, 2, and 4 mL, respec- Result = (V × C × DS)/(W × DA) × 100
tively, of Alcohol stock solution into three separate and
identical 25-mL volumetric flasks. To each flask add a V = volume of the Sample solution, 25 mL
5-mL portion of the Internal standard solution. Dilute DS = density of the Solution (g/mL)
with Diluent to volume, and mix thoroughly. The Stan- W = weight of Solution taken to prepare the
dard solutions contain 0.0006, 0.0012, and 0.0024 g/ Sample solution (g)
mL of alcohol (C2H5OH), respectively. DA = density of alcohol (g/mL)
Sample solution: Weigh an appropriate amount of So- Acceptance criteria: If present, 95.0%–105.0% of the
lution into a 25-mL volumetric flask, and pipet 5 mL of labeled amount of alcohol (C2H5OH)
the Internal standard solution into the flask. Dilute with
Diluent to volume, and mix thoroughly to obtain a solu- IMPURITIES
tion containing 0.0006–0.0024 g/mL of alcohol • LIMIT OF AMINES AND AMINE SALTS
(C2H5OH). Sample: A quantity of Solution equivalent to 5.0 g of
Chromatographic system benzalkonium chloride
(See Chromatography 〈621〉, System Suitability.) Analysis: Dissolve the Sample by heating carefully (e.g.,
Mode: GC on top of a steam bath with water as the steam source)
Detector: Flame ionization in 20 mL of a mixture of methanol and 1 N hydrochlo-
Column: 0.32-mm × 15-m glass or quartz capillary; ric acid VS (97:3). [NOTE—The mixed solution, however,
1.8-µm layer of phase G43 must not reach the boiling point.] Add 100 mL of iso-
NF Monographs

Temperatures propyl alcohol, and pass a stream of nitrogen slowly

Injection port: 250° through the solution. Gradually add 12.0 mL of 0.1 N
Detector: 320° tetrabutylammonium hydroxide VS while recording the
Column: See Table 2. potentiometric titration curve.
Acceptance criteria: If the curve shows two inflection
points, the volume of titrant added between the two
Table 2 points is NMT 5.0 mL, corresponding to NMT
Hold Time 0.1 mmol/g of amines and amine salts. If the curve
Initial Temperature Final at Final shows no point of inflection, the substance being ex-
Temperature Ramp Temperature Temperature amined does not comply with the test. If the curve
(°) (°/min) (°) (min) shows one point of inflection, repeat the test, but add
50 — 50 4 3.0 mL of a 25.0 mg/mL solution of dimethyldecy-
lamine in isopropyl alcohol before the titration. If after
50 10 70 0
the addition of 12.0 mL of the titrant, the titration
70 50 300 4 curve shows only one point of inflection, the substance
being examined does not comply with the test.
Run time: 14.6 min • LIMIT OF BENZYL ALCOHOL, BENZALDEHYDE, AND
Carrier gas: Helium (CHLOROMETHYL)BENZENE
Flow rate: See Table 3. [NOTE—Prepare the solutions immediately before use.]
Solution A: Dissolve 1.09 g of sodium 1-hex-
anesulfonate and 6.9 g of monobasic sodium phosphate
in water in a 1000-mL volumetric flask, adjust with

Official from May 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.
Accessed from 10.6.1.1 by JNJconsumer on Wed Jun 28 18:24:31 EDT 2017

NF 35 Official Monographs / Benzyl 7535

phosphoric acid to a pH of 3.5, and dilute with water (Chloromethyl)benzene: The response of the
to volume. (chloromethyl)benzene peak from the Sample solution
Solution B: Methanol is NMT 0.1 times that of the principal peak from Stan-
Mobile phase: See Table 4. dard solution A, corresponding to NMT 0.05%.
SPECIFIC TESTS
Table 4
• MICROBIAL ENUMERATION TESTS 〈61〉 and TESTS FOR SPECI-
Time Solution A Solution B FIED MICROORGANISMS 〈62〉: A solution containing less
(min) (%) (%) than 5.0% of benzalkonium chloride meets the require-
0 80 20 ments of the test for absence of Pseudomonas aeruginosa.
10 80 20 • ACIDITY OR ALKALINITY
Sample solution: Evaporate or dilute with carbon diox-
14 50 50
ide-free water to prepare a 50-mL solution of 10 mg/mL
35 50 50 of benzalkonium chloride in water.
36 20 80 Analysis: To the Sample solution add 0.1 mL of bromo-
55 20 80 cresol purple TS.
56 80 20 Acceptance criteria: NMT 0.5 mL of 0.1 N hydrochlo-
65 80 20 ric acid or 0.1 N sodium hydroxide is required to
change the color of the indicator.
Standard solution A: 0.25 mg/mL of USP Benzyl Alco-
hol RS in methanol ADDITIONAL REQUIREMENTS
Standard solution B: 0.075 mg/mL of USP Benzalde- • PACKAGING AND STORAGE: Preserve in tight containers,
hyde RS in methanol and prevent contact with metals.
Standard solution C: 0.025 mg/mL of USP Benzyl Alco- • LABELING: Label it to indicate the concentration of ben-
hol RS in methanol, prepared from Standard solution A zalkonium chloride, and to indicate the name and quan-
and methanol tity of the coloring agent added. The labeling also indi-
Sample solution: Determine the density of the Solu- cates the concentration of alcohol added.
tion. Dilute a quantity of the Solution equivalent to • USP REFERENCE STANDARDS 〈11〉
2.5 g of benzalkonium chloride with methanol to USP Benzaldehyde RS
50.0 mL. This solution contains 50 mg/mL of benzalko- USP Benzalkonium Chloride RS
nium chloride. USP Benzyl Alcohol RS
Chromatographic system
(See Chromatography 〈621〉, System Suitability.)
Mode: LC
Detector: UV 210 nm for benzyl alcohol and
(chloromethyl)benzene; UV 257 nm for benzaldehyde Benzethonium Chloride—see
Column: 4.6-mm × 15-cm; 5-µm packing L1 Benzethonium Chloride General Monographs
Column temperature: 30°
Flow rate: 1.0 mL/min
Injection volume: 20 µL
System suitability Benzoic Acid—see Benzoic Acid General
Samples: Standard solution A, Standard solution B,
Standard solution C, and Sample solution Monographs
[NOTE—See Table 5 for relative retention times.]

Table 5
.

Benzyl Alcohol
Relative
Portions of the monograph text that are national USP text,
Retention
and are not part of the harmonized text, are marked with
Name Time
symbols (◆◆) to specify this fact.
Benzyl alcohol 1.0
.

NF Monographs
Benzaldehyde 1.3
(Chloromethyl)benzene 2.4

Suitability requirements
Relative standard deviation: NMT 5.0% for benzyl C7H8O 108.14
alcohol, Standard solution A Phenylmethanol [100-51-6].
Signal-to-noise ratio: NLT 10 for the principal peak,
Standard solution C DEFINITION
Analysis Benzyl Alcohol contains NLT 98.0% and NMT the equivalent
Samples: Standard solution A, Standard solution B, of 100.5% of phenylmethanol (C7H8O).
Standard solution C, and Sample solution IDENTIFICATION
Calculate the content of (chloromethyl)benzene by mul- • ◆A. INFRARED ABSORPTION 〈197F〉: On undried specimen◆
tiplying the peak area of (chloromethyl)benzene by
.

1.3. [NOTE—The correction factor is used to adjust for ASSAY

baseline shift.] • PROCEDURE
Acceptance criteria Phenolphthalein solution: Dissolve 0.1 g of phenol-
Benzyl alcohol: The response of the benzyl alcohol phthalein in 80 mL of ethanol (96%), and dilute with
peak from the Sample solution is NMT that of the ben- water to 100.0 mL. To test for sensitivity, add 100 mL
zyl alcohol peak from Standard solution A, correspond- of carbon dioxide-free water to 0.1 mL of the Phenol-
ing to NMT 0.5%. phthalein solution. The solution is colorless. NMT 0.2 mL
Benzaldehyde: The response of the benzaldehyde of 0.02 M sodium hydroxide is required to change the
peak from the Sample solution is NMT that of the ben- color to pink.
zaldehyde peak from Standard solution B, correspond-
ing to NMT 0.15%.

Official from May 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.