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International Journal Of Scientific & Engineering Research, Volume 5, Issue 11, November -2014

ISSN 2229-5518

High-performance liquid chromatography

method for the analysis of Benzyl Alcohol
Vilas Khade, Sunil Mirgane

Abstract: A rapid economical, reproducible and simple direct method was developed and validated for determination of Benzyl
Alcohol using high performance liquid chromatography (HPLC). Benzyl Alcohol concentration estimated with isocratic elution using a
mixture of buffer and methanol and buffer (30: 70 v/v) within 10 min, Flow rate 1.00ml, and wavelength 215nm and column C18.
This method was suitable and validated for specificity, linearity, precision, accurate and ruggedness. The proposed method was
successfully applied in the determinate of raw material assay for Benzyl Alcohol .

Index Terms— Accurate & stability indicating chromatography method for Benzyl Alcohol
.
——————————  ——————————

also a precursor to a variety of esters, used in the

soap, perfume, and flavor industries. It is also used
1 INTRODUCTION: as a photographic developer.

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Benzyl alcohol is used as a bacteriostatic
preservative at low concentration in intravenous
medications, cosmetics and topical drugs. The use of
benzyl alcohol as a 5% solution has been approved
by the U.S. FDA in the treatment of head lice in
children older than 6 months and in adults.[4]
Benzyl alcohol is an aromatic alcohol with the
formula C6H5CH2OH. The benzyl group is often Benzyl alcohol is only a mild acute toxin with an
abbreviated "Bn", thus benzyl alcohol is denoted as LD50 of 1.2 g/kg in rats.[5] It oxidizes rapidly in
BnOH. Benzyl alcohol is a colorless liquid with a healthy individuals to benzoic acid, conjugated with
mild pleasant aromatic odor. It is a useful solvent glycine in the liver, and excreted as hippuric acid.
due to its polarity, low toxicity, and low vapor Very high concentrations can result in toxic effects
pressure. Benzyl alcohol is partially soluble in water including respiratory failure, vasodilation,
(4 g/100 mL) and completely miscible in alcohols hypotension, convulsions, and paralysis.
and diethyl ether. Benzyl alcohol is toxic to neonates, it is associated
with the gasping syndrome.[6][7].
Benzyl alcohol is produced naturally by many
plants and is commonly found in fruits and teas. It Benzyl alcohol has been reported to cause skin
is also found in a variety of essential oils including allergy.[8].
jasmine, hyacinth, and ylang-ylang.[1]
It is also one of the chemical compounds found in Benzyl alcohol is severely toxic and highly irritating
castoreum. This compound is gathered from the to the eye.[5] Pure benzyl alcohol produces corneal
beaver plant food.[2] necrosis.[9].
Benzyl alcohol is prepared by the hydrolysis of
benzyl chloride using sodium hydroxide: Benzyl alcohol is not considered to be a carcinogen
C6H5CH2Cl + NaOH → C6H5CH2OH + NaCl and no data are available regarding teratogenic or
It can also be prepared via a Grignard reaction by reproductive effects.[5]
reacting phenylmagnesium bromide (C6H5MgBr)
with formaldehyde, followed by acidification No chromatography method is available for
Benzyl alcohol is used as a general solvent for inks, determination Benzyl Alcohol assay in raw material.
paints, lacquers, and epoxy resin coatings [3].It is At present all pharmacopeia i.e., United
state Pharmacopeia USP 34 [10],

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International Journal Of Scientific & Engineering Research, Volume 5, Issue 11, November -2014

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British Pharmacopeias 2014 [11] and Indian response of the peaks due to Benzyl alcohol.
Pharmacopoeia [12] having potentiometric
titrimetry methods. 2.7 System suitability:
The system is suitable for analysis if,
2 EXPERIMENTAL: Relative standard deviation for five replicate
2.1 Reagent & chemical: injections of both standard preparations is not more
All the employed solvent were of HPLC grade and than 2.0%.
were obtained from Merck. All other chemicals
were analytical-reagent grade and deionized water 2.8 Calculation :
was used to prepare solution. Calculate the amount of Benzyl Alcohol in %w/w
HPLC system consisting pump, UV detector, auto by using following
sampler software for the detection. Column used 250
x 4mm, RP 18, 10 microns. Chromatographic
condition is as follows. Formula:

2.2 Chromatographic condition: Sample Weight of % Potency of

a) Flow rate : 1.0 ml /min. Area Standard in mg standard (as such)
b) Detection : 215 nm. = -----------------x---------------------x-------------------x 100
c) Application : 20 µl Standard Weight of 100
d) Run Time : 10 min, Area Sample in mg

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2.3 Mobile Phase: 3 METHOD VALIDATION [13], [14] [15]
Mix Methanol and 0.02M Phosphoric acid (70:30)
filter through a membrane filter of 0.2 micron and 3.1 Precision:
degas it with ultrasonic bath for 5 to 10 minutes. The precision was tested with 5 repeated injection of
Benzyl Alcohol standard solution. The relative
Preparation of 0.02 M Phosphoric Acid: standard deviation (RSD) is less then 2.0% . The
Dilute 1.36 ml Orthophosphoric acid to 1000 ml with reproducibility of the chromatographic separation
water. was very good as shown by the very narrow
2.4 Standard Preparation: window of the retention time.
Weigh accurately about 20.0 mg Benzyl Alcohol in
100 ml volumetric flask. Add 50 ml of the mobile 3.2 Specificity:
phase. Sonicate it for 5 minutes and dilute up to Initially blank sample without Benzyl Alcohol inject.
mark with mobile phase. No peak observed in blank solution. Benzyl Alcohol
Standard Inject and peck was observed at 4.0
Pipette out 25 ml of above solution in 100 ml retention time (RT). No interference of any peck and
volumetric flask and make up the volume to the pack purity is 100%.
mark with Mobile phase and mix.
3.3 Accuracy:
2.5 Sample Preparation: Initially blank sample without Benzyl alcohol were
Weigh accurately about 20.0 mg Benzyl Alcohol in injected for the know any interference. working
100 ml volumetric flask. Add 50 ml of the mobile standard was used for the accuracy testing.
phase. Sonicate it for 5 minutes and dilute up to Compared sample Benzyl alcohol against working
mark with mobile phase. standard. The retention time of Std. is 4.033 &
sample retention time is 4.067. Assay is 99.99%
Pipette out 25 ml of above solution in 100 ml against the standard.
volumetric flask and make up the volume to the
mark with Mobile phase and mix. 3.4 Recovery:
The recovery was tested & it observed that recovery
2.6 Procedure for injection: is about 99.86%
Separately inject equal volume of about 20µl of
blank, standard solution in five replicate and sample 3.5 Linearity:
solution in duplicate, into the equilibrated HPLC Under the above-described optimum conditions, the
system, record the chromatograms and measure the calibration curve obtained with standard showed

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International Journal Of Scientific & Engineering Research, Volume 5, Issue 11, November -2014

ISSN 2229-5518

good linear relationship in the interval 20 pm to 80 [12] Indian Pharmacopoeia 2010.

ppm. The correlation coefficient was observed [13] International Conference on Harmonization
0.9994 for the calibration curve. (ICH ) Harmonised Tripartite Guideline (1996) Step
4 version, Parent Guideline dated 27 October 1994,
3.6 Robustness: (Complementary Guideline on Methodology. dated
Flow rate change: 6 November 1996 incorporated in November 2005)
Flow rate was change 0.8ml/min, 1.0ml/min, [14] United state Pharmacopeia USP 34, Validation
1.2ml/min and check the effect on assay of Benzyl of Compendial Procedure 1225.
Alcohol . No variation was observed in assay due to [15] British Pharmacopeias, Validation of Analytical
this change. Procedure SC III F Volume V.

Wavelength change:
Wave length was change 210nm, 215nm, 220nm
check the effect on assay of propyl paraben. No
variation was observed in assay due to change in
wavelength.

4 CONCLUSION:
This work presents a simple and validated HPLC
method for the determination of Benzyl Alcohol.
The method was validated showing satisfactory

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data for all parameters tested. Thus, it offers
advantages over other analytical methods due to its
rapidity, simplicity and lower cost.

REFERENCE:
[1] The Merck Index: An Encyclopedia of Chemicals,
Drugs, and Biologicals (11th ed.), Merck, 1989,
ISBN 091191028X , 1138.
[2] The Beaver: Its Life and Impact. Dietland Muller-
Schwarze, 2003,
[3] Benzyl alcohol, chemicalland21.com .
[4] . Prescribing Information for Ulesfia Lotion,
Sciele Pharmaceuticals, Inc., April 2009, retrieved
2009-08-29 .
[5] Friedrich Brühne; Elaine Wright (2007), "Benzyl
Alcohol", Ullmann's Encyclopedia of Industrial
Chemistry (7th ed.), Wiley, pp. 7–8
[6] Carl R. Baum (2008), "Examples of mass
exposures involving the pediatric population", in
Jerrold B. Leikin; Frank P. Paloucek, Poisoning and
Toxicology Handbook (4th ed.), Informa, p. 726
[7] Juan Gershanik et al. (1982), "The gasping
syndrome and benzyl alcohol poisoning.", N Engl J
Med307: 1384–8, doi:10.1056/nejm198211253072206
[8] EJ Curry; EM Warshaw (2005), "Benzyl alcohol
allergy: importance of patch testing with personal
products.", Dermatitis (16): 203–8
[9] Swarupa G Kulkarni; Harihara M Mehendale
(2005), "Benzyl Alcohol", Encyclopedia of
Toxicology 1 (2nd ed.), Elsevier, pp. 262–264
[10] United state Pharmacopeia USP 37, NF 32 2014.
[11] British Pharmacopeias 2014.

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