DRUG INFORMATION

NAME OF THE DRUG: N-acetyl L- cysteine

STRUCTURE:

IUPAC NAME: (2R)-2-acetamido-3-sulfanylpropanoic acid

SYNONYMS :acetein, acetylcysteine, N-acetylcysteine, N-acetyl-N-cysteine, N- acetyl-3mercaptoalanine, airbron, broncholysin, fluimucetin, fluimucil, flumicil, inspir, mercapturic acid,
mucolyticum, mucolyticum lappe, mucolytikum lappe, mucomyst, mucosolvin, NAC, NAC-TB, NSC
111180, parvolex, respaire.

MOLECULAR FORMULA: C5H9NO3S

MOLECULAR WEIGHT: 163.2 (anhydrous)

PHYSICAL PROPERTIES
1) Appearance:
EUROPEAN
PHARMACOPOEIA
A white crystalline
powder or colourless
crystals

UNITED STATES
PHARMACOPOEIA
-

SIGMA ALDRICH
White to white with
light yellow cast
powder

2) Melting Point:
EUROPEAN
PHARMACOPOEIA
104OC - 1100C

UNITED STATES
PHARMACOPOEIA
-

SIGMA ALDRICH
109C -110C

3)Specific optical rotation

EUROPEAN
PHARMACOPOEIA
+21.0 to +27 .0

UNITED STATES
PHARMACOPOEIA
+21 to +27

SIGMA ALDRICH
Optical rotation:
+5(c = 3% in
water)

4)pH :
EUROPEAN
PHARMACOPOEIA
2.0 to 2.8(0.125g/ml)

UNITED STATES
PHARMACOPOEIA
2.0 to 2.8(1in 100)

SIGMA ALDRICH
-

5) Odour : slight acetic odour

6) Taste: characteristic sour taste
7) pKa - 3.24 (carboxylic acid moiety), 9.52 (SH group)
8) logP - -0.66
9) Solubility
EUROPEAN
PHARMACOPOEIA
Soluble in water and
alcohol
insoluble in methylene
chloride

UNITED STATES
PHARMACOPOEIA
-

SIGMA ALDRICH
soluble 1 in 8 of water
and 1 in 2 of ethanol
insoluble in chloroform
and ether

 Soluble in hot isopropyl alcohol, methyl acetate, and ethyl acetate

RELATED SUBSTANCES
A) L-cystine (EP impurity A,USP impurity A)

B) L-cysteine ( EP Impurity B)
L-cysteine hydrochloride (USP impurity B)

C) N,N-diacetyl L-cystine(EP impurity C,USP impurity C)

D) N,S-diacetyl L-cysteine(EP impurity D,USP impurity D)

JOURNAL ARTICLES AND MONOGRAPHS

Journal name

Author

Article name

Comments

Analytical
methods

Aline Ferreira ourique et

al

A LC UV method to assay
N- acetyl cysteine without
derrivitization :analysis of
pharmaceutical products

Column :phenomenex luna C18 (250x

4.60 mm,5 microns,100A0 )
Mobile phase :KH2PO4(0.05M):ACN
(95:5)(pH adjusted with 0.095%
phosphoric acid )
Injection volume -100l
Flow rate :1.3ml/min
Wavelength :214 nm
Run time -10 mins
Retention time :6 mins
Temperature :251C

Journal of
chromatography

B.toussaint et al

Quantitative analysis of N
acetyl cysteine and its
pharmacopoeial
impurities in a
pharmaceutical
formulation by liquid
chromatography UV
detection mass
spectrometry.

Column :platinum EPS C18

(150x2.1mm,5 microns )
Mobile phase :ACN :NH3-formic acid
(7/12mM)(2:98v/v)at pH3
Flow rate -0.3ml/min
Wavelength :210 nm
Injection volume -10 l
Temperature -25C

- it includes all the four

pharmaceutical impurities as per EP
Journal of
pharmaceutical
and biomedical
analysis

J Farquhar et al

A reversed phase high

performance liquid
chromatographic assay for
the determination of Nacetyl cysteine in aqueous
formulations

Column :
C18spherisorb(200mm5mm,10m)
Mobile phase :0.5%m/v aqueous solution
of sodium perchlorate at pH2
Flow rate -2ml/min
Wavelength -215nm
Temperature -60C

International
journal of drug
development and
research

Sheikh sana et al

Development and
validation of RP-HPLC
method for the estimation
of N-acetyl cysteine in
wet cough syrup

Using internal standard solutions

-0.4mg/ml l-tyrosine in dil ortho
phosphoric acid (pH2)
The method is capable for the
simultaneous determination of
acetyl cysteine and N,N diacetyl
cysteine (impurity C)
from the degradation studies
carried out by exposing the
aqueous formulation (pH6) to
atmospheric oxygen for
4,20,30,37 for 12 months reports
the formation of N,N diacetyl
cystine (impurity C) N,S diacetyl
cystiene(imp D) ,L-cysteine(imp
B) and an impurity 4 (structure
given in the article )
The retention time of the impurity
4 has been provided but its
formation nor its structure
elucidation has been discussed .

Column :waters symmetry C18

(150x4.6mm i.d.,3.5microns )

Mobile phase :KH2PO4(pH3):ACN

Time
0
5
10
5

A
95
95
20

B
5
5
80

15
20

95
95

5
5

Flow rate -0.8ml/min

Run time -20 mins
forced degradation studies were
carried under acid,base,peroxide
,sulfite ,heat and light .
The four pharmacopoeial
impurities had been identified in
the degraded sample .
Other unknown impurity peaks in
addition to the pharmacopoeial
impurities were also observed .
American journal
of health systems
pharmacy

Alexander L.Fohl et al

Stability of
extemporaneously
prepared acetyl cysteine
1% and 10 %solutions for
treatment of meconium
ileus

Column -1503.9mm,4m
Mobile phase 0.05M KH2PO4(pH3)
Wavelength -210 nm
Flow rate -1ml /min

Journal of
pharmacy practice
and research

The phung To et al

Stability of a formulated
N-acetyl cysteine capsule
for prevention of contrast
induced nephropathy

Degradation samples were

prepared in acidic ,alkaline
,oxidative conditions .The
retention time for the degradation
products have been provided but
whether these match to
pharmaceutical impurities is
unclear .
Stability samples (1% aqnd 10 %)
were prepared and stored at room
temperature and samples were
withdrawn at 7,14,30,60 and 90
days .A significant decrease in
percentage of NAC has been
reported at 90 days but no
information regarding the
degradtion products has been
provided .
Hence it is unclear if the
degradation products match with
the pharmacopoeial impurities and
whether new impurities are
formed .
Column
AllsphereODS2(250mm4.6mm,5)
Mobile phase

-0.05MKH2PO4(pH3):ACN(98:2)
Wavelength -214nm
Flow rate -1.5ml/min
Run time -9 minutes

American journal
of health systems
pharmacy

Rivka Siden et al

Stability of a flavoured
formulation of acetyl
cysteine for oral
administration

Capsules were stored at 2-8C,

18-25C,and 40C with and
without desiccant at ambient and
high humidity .
The article reports that in the high
temperature high humidity
conditions there was significant
loss in percentage of NAC but it
could not be accounted for in
terms of mass balance production
of NAC impurities .It assumes
that the imbalance can be due to
the inaccurate recovery of acetyl
cysteine from the degraded
capsules or NAC degraded into
other break down products that
were undetectable by the HPLC
method used in the study .

Column-250mm4.6mm,5
Mobile phase -0.05MKH2PO4 (pH3)
Wavelength -210 nm
Flow rate 1ml/min
Retention time -9.48 mins

Strawberry flavoured acetyl

cysteine solution was prepared
and stored at room temperature
and in refrigerated conditions
.samples were withdrawn at
0,7,14,21,28 and 35 days . There
is a decrease in percentage of
acetyl cysteine but there is no
mention whether or not
degradation products have been
formed .
The forced degradation samples

Peritoneal dialysis
international

Eun-Young Seo et al

Stability of N-acetyl
cysteine in peritoneal
dialysis solution

were prepared under

oxidative,acidic and alkaline
conditions .The article reports the
retention time of N,N diacetyl
cysteine(acetyl cysteine impurity
C) which is the oxidative
degradation product.
It reports the presence of other
degradation peaks but it is unclear
as to whether the degradation
peaks correspond to the
pharmacopoeial impurities or not .
Column phenomenex C18
Mobile phase -0.05MKH2PO4 (pH3)
Flow rate -1.5ml/min
Wavelength -214 nm
Retention time -5.4 mins

Annals of
emerging
medicine

William H.Dribben et al

Stability and
microbiology of inhalant
N-acetyl cysteine used as
an intravenous solution
for the treatment of
acetaminophen poisoning

The solutions of NAC in

peritoneal dialysis solution were
kept at 4C and at room
temperature .Samples were
withdrawn at
0,1,3,7,15,30,60,90,120,150,180
days .A decrease in percentage of
NAC has been reported but no
information whether or not
degradation products are formed
has been provided .
Solutions of NAC were forcefully
degraded under acidic and
alkaline conditions .But no
information is provided regarding
the degradation products .
So it is unclear whether the
impurities generated match with
the pharmacopoeial impurities .
USP method for the assay of acetyl
cysteine
Solutions of inhalent NAC were
compounded and stored at
25C/60%RH,40C/75%RH and
at ambient conditions .the samples
were withdrawn at
0,4,8,12,24,36,48,60 and 72 hrs .
A decrease in percentage of NAC

Journal of the
Chilean Chemical
Society

Siddiquiet al

Iodate oxidation of Nacetyl L-cysteine

:application in drug
determination and
characterization of its
oxidation and degradation
products by mass
spectrometry

has been reported .No information

whether or not impurities has been
generated has been provided .
The article reports that the
solution which was kept at
accelerated conditions did not
degrade at those conditions hence
it is limited to drawing
conclusions on the effects of these
accelerated conditions on stability
hence it is unclear whether the
impurities if generated will match
with the pharmacopoeial
impurities or not .
A kinetic spectrophotometric
method based on initial rate
measurement of the oxidation of
of N-acetyl cysteine with iodate
has been developed for the
determination of N-acetyl cysteine
.The reaction product was
characterized using mass
spectrometry .The m/z ratio was
326 which indicated the formation
of disulphide acetyl cysteine
The degradation products of
acetyl cysteine was also prepared
and characterized
1.5310-4 M NAC in presence
of
1.2510-2 M sodium hydroxide
was heated at 100Cat 30
minutes .In this condition the
acetamide group breaks away
from the parent compound
leaving behind a compound with
m/z ratio 103 .Since acetamide
is highly unstable it gets
converted to acetic acid having
m/z ratio of 59
1.5310-4 M NAC was heated
with
5.010-4M hydrochloric acid for
30 minutes at 100C.The acid
degradation product has a mass
no of 121 and from the mass
spectra formation of cysteine
was predicted
100mg of acetyl cysteine was
heated at 100C for 30 minutes

Analytica Chimica
Acta

Xinyi Wang Ruoyun et al

N-acetylcysteine induced
quenching of red
fluorescent
oligonucleotide-stabilized
silver nanoclusters and the
application in
pharmaceutical detection

Journal of
chromatography
and biomedical
applications

Nuran Ercal et al

High-performance liquid
chromatography assay for
N-acetylcysteine in
biological samples
following derivatization
with N-( 1pyrenyl)maleimide

10

and from the heated sample

1.5310-4 M NAC was prepared
.The mass spectrometric
analysis revealed a compound with mass
no 119.04
1.5310-4 M NAC was
subjected to peroxide
degradation by subjecting
heating the parent compound in
presence of 5ml of 3% hydrogen
peroxide for 30 mins at
100C.The mass spectra had
revealed a major compound is
formed at m/z 160.
The degradation products
prepared under iodate/iodide
conditions is N,S diacetyl
cysteine which is acetyl cysteine
impurity D and has been
reported by few articles as an
impurity formed during degradation studies .
The degradation product under
acidic condition -cysteine
correspond to impurity B as per
EP and has been reported by a
few articles as a manufacturing
impurity as well as a degrdation
product .
The degrdation products under
alkaline ,peroxide and heat
conditions does not match with
the pharacopoeial impurities and
have not yet been reported in the
stability related articles .
A new, simple and sensitive method for
determination of N-acetylcysteine (NAC)
based on quenching of the red
fluorescence of oligonuleotide-protected
silver nanoculsters (Ag NCs) was
described This method was further used
for the assay of N-acetyl cysteine in
granules.
Column C18 (100x 4.6 mm ,3)( (Astec,
Whippany, NJ, USA)
Mobile phase water: acetonitrile (50:50,
v/v) (pH 3.75)
Flow rate - 0.45 ml/min.

Excitation wavelength - 330 nm

Emission wavelength of 380 nm.
Scientia
Pharmaceutica

Rim Haggag et al

Derivatization with 4Chloro-7-nitro-2,1,3benzoxadiazole for the

spectrophotometric and
differential Pulse
polarographic
determination of
acetylcysteine and
captopri

Chromatographia

V. Cavrini et al

HPLC Determination of
Thiol Drugs in
Pharmaceutical
Formulations Using
Ethacrynic Acid as a
Precolumn Ultraviolet
Derivatization Reagent

Analytical Letters

W. Baeyens et al

Hplc Determination of NAcetylcysteine in

Pharmaceutical
Preparations After PreColumn Derivatization
With ThiolyteR MB Using
Fluorescence Detection

Injection volume - 20l

Acetylcysteine and captopril were
determined based on their reaction with 4chloro-7-nitro-2, 1, 3-benzoxadiazole
(NBD-Cl) in the presence of sodium
tetraborate in absolute methanol. The
yellow coloured products obtained were
measured spectrophotometrically at 417
and 420 nm for acetyl cysteine and
captopril respectively and by differential
pulse polarography at -872 and -1007 mV
for acetyl cysteine and captopril
respectively .The methods were validated
and used for the determination of drugs in
pharmaceutical preparations.
HPLC method has been developed for the
determination of aliphatic thiol drugs,
such as N-acetyI-L-cysteine, captopril and
mer- captopropionylglycine in
pharmaceutical formulations. The
procedure involves a precolumn
derivatization of the thiol drug with
ethacrynic acid followed by reversedphase HPLC separation and UV detection.
The method is reliable for the quality
control of commercial dosage forms of the
examined thiol drugs.
Column- Lichrosorb C18 ( 250 x 4.6 mm ,
10)
Temperature - 22 C
Flow rate -2ml/min
Mobile phase -acetic acid 1 % (v/v) 95
ml : acetone 5 ml.
Excitation wavelength - 380 nm
Emission wavelengths-470 nm

Acta
chromatographica

11

Y. Vander Heyden et al

Development and
validation of an HPLC
method with post-column
derivatization for assay of
N-acetylcysteine in
plasma

Column-LiChrosorb C-18 ( 250 mm 4.6

mm , 5 m)
Mobile phase - 1% acetonitrile (ACN) in
phosphate buffer (pH 3.0)

flow rate- 1.0 mL min1.

Excitation wavelength - 365 nm
Emission wavelength -442 nm
Science Asia

Elham Anwer Tahaa et al

Kinetic
Spectrophotometic
Determination of
Acetylcysteine and
Carbocisteine in Bulk
Powder and in Drug
Formulations

International
Journal of
Pharmaceutical
and
Phytopharmacolo
gical Research

Nitin S et al

International
Journal of
Analytical
Chemistry

Lea Kukoc-Modun et al

Development and
validation of
spectroscopic method for
simultaneous estimation
of acebrophylline and
acetylcysteine in capsule
dosage form
Spectrophotometric
Determination of NAcetyl-L-Cysteine and N(2-Mercaptopropionyl)Glycine in Pharmaceutical
Preparations

Molecules

Josipa Giljanovi et al

12

Flow Injection
Spectrophotometric
Determination of NAcetyl-L-cysteine as a
Complex with
Palladium(II)

Kinetic spectrophotometric method for the

determination of acetylcysteine and
carbocisteine is described.The method is
based on the reaction between the drugs
and 4-chloro-7-nitrobenzo-2-oxa 1, 3
-diazole (NBD-Cl) in an alkaline
medium.the absorbance was measured at
424 nm at ambient temperature (250C 5)
for a fixed time of 30 minutes for acetyl
cysteine , and at 468 nm for a fixed time
of 15 minutes at 70 0C for
Carbocisteine.the method was successfully
applied for the determination of the two
drugs in bulk powder, in pharmaceutical
formulations as well as in the presence of
their related substances .
A simple, economic, sensitive, accurate
and reproducible spectroscopic method
has been developed and validated for the
simultaneous estimation of Acebrophylline
and Acetylcysteine in capsule dosage form
by Simultaneous equation method.
The proposed equilibrium method for the
determination of N-acetyl-L-cysteine
(NAC) and N-(2mercaptopropionyl)glycine (MPG) is
based on a coupled two-step redox and
complexation reaction. In the first step,
Fe(III) is reduced to Fe(II) by NAC or
MPG. Subsequently, Fe(II) is complexed
with 2,4,6-tripyridyl-s- triazine
(TPTZ).The method was successfully
applied to quantify NAC and MPG in
pharmaceutical preparations.
A flow-injection analysis with spectrophotometric detection, suitable for the
determination of N-acetyl-L-cysteine
(NAC) is described . NAC and Pd2+ form
complexes of Pd2+:NAC molar ratios of
1:1 and 1:2.The proposed method was
compared with the classic
spectrophotometric determination of
NAC, using the same reagent, PdCl2.

Journal of .
Brazillian
Chemical .
Society

Willian T. Suarez et al

Flow Injection
Turbidimetric
Determination of
Acetylcysteine in
Pharmaceutical
Formulations Using Silver
Nitrate as Precipitant
Reagent

Instrumentation
Science &
Technology

Paraskevas D et al

A Green Hplc Method For

The Determination Of NAcetylcysteine Using
Post-Column
Derivatization With
Methyl-Propiolate

A simple, accurate and precise flowinjection turbidimetric procedure is

reported for the determination of
acetylcysteine in pharmaceutical
formulations. The procedure is based on
the precipitation of acetylcysteine with
silver nitrate solution in acid medium and
the insoluble salt produced was monitored
at 410 nm.
Column - C18( 150 4.6 mm i.d.,5 m )(
Prevail )
Mobile phase - 0.05 % v/v CH3COOH + 1
mmol L1 EDTA in water
Wavelength - 285 nm
Flow rate -1ml/min

Journal of
Chemistry

Shaesta quyoom et al

Potentiometric and UV
Spectral Studies of Binary
and Ternary Complexes of
Some Metal Ions with NAcetylcysteine and Amino
Acids

International
journal of
electrochemical
science

Acelino Cardoso de S et
al

Determination of NAcetylcysteine by Cyclic

Voltammetry Using
Modified Carbon Paste
Electrode with Copper
Nitroprusside Adsorbed
on the 3
Aminopropylsilica.

Biopharmaceutics
& drug disposition

Bernard Gabard

Endogenous plasma Nacetylcysteine and single

dose oral bioavailability
from two different
formulations as
determined by a new

13

The formation constants of the binary 1:1

and 1:2 and 1:1:1 ternary complexes
complexes of Cu(II), Zn(II), Cd(II),
Hg(II), and Pb(II) with N-acetylcysteine
and some biologically important amino
acids as secondary ligands have been
determined potentiometrically in aqueous
mediumat 25C. The formation constants
of the 1:1 complexes were found to be
higher than 1:2 complexes and the metal
ions follow the order Hg(II) >Cu(II)
>Cd(II) >Zn(II).
Copper nitroprusside was formed on
aminopropylsilica silica gel surface .The
cyclic voltammogram of CuNPSD were
found to exhibit two redox couples with
(E)1 = 0.34 V; (E)2 = 0.76 V.The second
redox process ((E)2 presented by the
graphite paste electrode with SiCuNP
shows electrocatalytic activity for the
oxidation of N-acetylcysteine. The linear
range for the determination of Nacetylcysteine was found between 9.9
10-5 and 8.9 10-4 mol L-1 showing a
detection limit of 4.18 10-5 mol L-1
Column -Nucleosil 120 C18 3 m (80 x 4
mm)
Excitation wavelength- 325 nm,
Emission wavelength - 415 nm

analytical method.

Mobile phase : Ethano l:H3P04 (0.1 M)

(4:96)

Acta Chimica
Slovenica

Ana Luiza de Toledo

Fornazari et al

Flow Injection
Spectrophotometric
System for N-Acetyl-LCysteine Determination in
Pharmaceuticals

Journal of
Chemical research

Waseem Amir

Flow-injection
Determination of
Cysteine, N-Acetyl
Cysteine and Glutathione
in Pharmaceuticals via
Potassium FerricyanideFe(III)
Spectrophotometric
System

Journal of
Chromatography

C. Celma et al

Determination of Nacetylcysteine in human

plasma by liquid
chromatography coupled
to tandem mass
spectrometry

In this method N-acetylcysteine in

plasma is reduced using tributylphosphine followed by postHPLC column derivatisation with
o-phthalaldehyde
A crossover study was done to
compare the bioavailability of two
different formulations of Nacetylcysteine.
The drug was detected in plasma
for up to 12 h after administration
of a single oral dose
The oxidation of N-acetyl-L-cysteine by
iron(III) is performed and the iron(II)
produced was determined
spectrophotometrically as a stable
tris(1,10-phenantroline)iron(II) complex at
510 nm
A simple, rapid and economical flow
injection spectrophotometric method for
the determination of cysteine, N-acetyl
cysteine and glutathione in pharmaceutical
formulations was established with Fe(III)/
ferricyanide reduction and soluble
Prussian blue detection system. The
calibration graphs are linear in the
concentration ranges of (1100)106
mol/L for cysteine and N-acetyl cysteine,
and (150)106 mol/L for glutathione
Column -. Kromasil C18 column (5034.6
mm ,5 microns )
Mobile phase - acetonitrile : water (70:30)
Flow rate - 1 ml/min.

14

The analytical method consists of

plasma digestion with
dithiothreitol in order to reduce all
the oxidized forms of Nacetylcysteine, and extraction with
ethyl acetate followed by
determination of levels by an LC
MSMS method

Journal of
Chromatography
B:Biomedical
applications

Longo A.,Toro M.

Determination of Nacetylcysteine in human

plasma by gas
chromatographymass
spectrometry

International
Journal of
Electrochemical
Science

Ante Prki et al

Direct Potentiometric
Determination of Nacetyl-L-cysteine (NAC)
in Real Samples by Using
home made Iodide ISE

Ghannam S.,Brashy A.et

al

Fluorimetric
determination of some
thiol compounds in their
dosage forms

Il Farmaco

NAC was extracted from plasma with

ethyl acetate and derivatized in two steps
with 2-praponol and pentafluoropropionic
anhydride. The volatile derivative
obtained was ideal for gas
chromatographicmass spectrometric
analysis.
Iodide ion selective electrode membrane
was made of
AgI:Ag2S:PTFE( Polytetrafluoroethylene )
= 1:1:2. The Proposed method was used
the for determination of NAC in acetic
buffer, pH 5 without pre-treatment of
pharmaceuticals. The determination is
based on the reaction between NAC and
Ag+ from electrode membrane. The
method has a linear response range for
NAC from 2105 to 1102 mol L1 with
limit of detection of 7.8106 mol L1.
Three pharmaceutical compounds
containing thiol groups namely, captopril,
d-penicillamine and N-acetylcysteine
were detrmined. The drugs are reacted
with 1,2-naphthoquinone-4-sulfonic acid.
The latter is reduced to 1,2dihydroxynaphthalene-4-sulfonic acid
which has a maximum fluorescence
intensity at 480/318 nm (Em/Ex).the
method has a range of 0.54.5 g ml1
with minimum detectability 0.05 g ml1

MONOGRAPHS
European pharmacopeia

Test for related substances

Column C18 (0.25Mx 4mm .)

Note the related substance LC method undergoing improvement

Mobile phase water R :ACN (97:3) (mix and

adjust
to pH3)
Wavelength -220nm
Injection volume-20l
Retention time for acetyl cysteine -6.4 minutes
L cysteine-2.2 mins
L-cysteine-2.4mins
Acetyl cysteine impurity C-12 mins
Acetyl cysteine impurity D-14 mins

15

The sample and standard solutions are prepared

in 0.01Mhydrochloric acid
United states pharmacopoeia

Assay of acetyl cysteine

Column :C18 (30cmx3.9 mmi.d.)
Mobile phase 0.05M potassium dihydrogen
phosphate buffer (pH3)

Note : Monograph being revised

Flow rate -1.5 ml/min
Injection volume -10l
Wavelength -214nm
Internal standard DL-phenyl alanine

United states pharmacopoeia

Relative retention time -6.4 minutes

Internal standard DL-phenylalanine
Sample ,standard and internal standard
solutions are prepared in sodium
metabisulfite solution (1in 2000)

Assay of acetyl cysteine solution

Column :C18 (30cmx3.9 mmi.d.)
Mobile phase 0.05M potassium dihydrogen
phosphate buffer (pH3)
Flow rate -1.5 ml/min
Injection volume -10l
Wavelength -214nm
Internal standard DL-phenyl alanine

USP Medicines Compendium

Note Monographs proposed for development

16

Relative retention time -6.4 minutes

Internal standard DL-phenylalanine

Sample ,standard and internal standard

solutions are prepared in sodium metabisulfite
solution (1in 2000)
Acetyl cysteine capsules
Acetyl cysteine for injection
Acetyl cysteine for oral solution
Acetyl cysteine inhalation solution
Acetyl cysteine injection
Acetyl cysteine oral solution
Acetyl cysteine tablets
Acetyl cysteine and Cefexime tablets
Acetyl cysteine and Isoproterenol inhalation

solution

British pharmacopoeia (test for related substances of acetyl cysteine)

Column -0.25Mx4mm i.d.

Stationary phase octadecyl silyl silica gel ,5
microns
Mobile phase water :ACN (97:3)
Flow rate -1ml /min
Wavelength -220nm
Run time -30 minutes
Retention time -6.4 minutes

BIOCHEMISTRY AND PHARMACOKINETICS

NAC is a sulfhydryl-containing compound rapidly absorbed into various tissues following an
oral dose, deacetylated and metabolized in the intestines and liver, and its metabolites
incorporated into proteins and peptides. Peak plasma levels of NAC occur approximately one
hour after an oral dose; at 12 hours post-dose it is undetectable in plasma. Despite a relatively
low bioavailability of only 4-10 per cent, oral administration of NAC appears to be clinically
effective. The biological activity of NAC is attributed to its sulfhydryl group; while its
acetylsubstituted amino group affords it protection against oxidative and metabolic
processes.NAC administration is an effective method of increasing plasma glutathione (GSH)
levels, as incorporation of cysteine into GSH appears to be the rate-limiting step in GSH
synthesis.
MECHANISMS OF ACTION
NACs effectiveness is primarily attributed to its ability to reduce extracellular cystine to
cysteine, and as a source of sulfhydryl groups. NAC stimulates glutathione synthesis, enhances
glutathione-S-transferase activity, promotes liver detoxification by inhibiting xenobiotic
biotransformation, and is a powerful nucleophile capable of scavenging free radicals. NACs
effectiveness as a mucolytic agent results from its sulfhydryl group interacting with disulphide
bonds in mucoproteins, with mucus subsequently being broken into smaller, less viscous units.

17

REFRENCES
1. Cavrini V , Gatti R. HPLC Determination Of Thiol Drugs In Pharmaceutical Formulations Using
Ethacrynic Acid As A Precolumn Ultraviolet Derivatization Reagent . Chromatographia 1987;
23(9): 680.
2. Paraskevas D. Tzanavaras. A Green Hplc Method For The Determination Of N-Acetylcysteine
Using Post-Column Derivatization With Methyl-Propiolate. Instrumentation Science &
Technology 2012.; 40 (2): 150-160
3. Quyoom S, Khan B.. Potentiometric And Uv Spectral Studies Of Binary And Ternary Complexes
Of Some Metal Ions With N-Acetylcysteine And Amino Acids . Journal Of Chemistry 2009;
6(S1): S117-S122
4. Cardoso A., Paim L,. Determination Of N-Acetylcysteine By Cyclic Voltammetry Using
Modified Carbon Paste Electrode With Copper Nitroprusside Adsorbed On The 3
Aminopropylsilica. International Journal Of Electrochemical Science 2011; 6(): 3754 - 3767.
5. Fornazari A,Suarez W. Flow Injection Spectrophotometric System For N-Acetyl-L- Cysteine
Determination In Pharmaceuticals. Acta Chimica Slovenica 2005; 52(): 164167.
6. Prki A, Giljanovi J. Direct Potentiometric Determination Of N-Acetyl-L-Cysteine (NAC) In
Real Samples By Using Home Made Iodide ISE . International Journal Of Electrochemical
Science 2011; 6: 5388 5395
7. Giljanovi J Brkljaa M,. Flow Injection Spectrophotometric Determination Of N-Acetyl-LCysteine As A Complex With Palladium(II) . Molecules 2011; 16: 7224-7236
8. Celma C ,Allue A. Determination Of N-Acetylcysteine In Human Plasma By Liquid
Chromatography Coupled To Tandem Mass Spectrometry . Journal Of Chromatography 2000;
870 : 1322
9. Wang X, Lin R. N-Acetylcysteine Induced Quenching Of Red Fluorescent OligonucleotideStabilized Silver Nanoclusters And The Application In Pharmaceutical Detection. Analytica
Chimica Acta 2013: 1-23.
10. Haggag R,Belal S.,Shaalan R. Derivatization With 4-Chloro-7-Nitro-2,1,3-Benzoxadiazole For
The Spectrophotometric And Differential Pulse Polarographic Determination Of Acetylcysteine
And Captopril . Scientia Pharmaceutica 2008; 76: 3348 .
11. Jadhav N,Lalitha K. DEVELOPMENT AND VALIDATION OF SPECTROSCOPIC METHOD
FOR SIMULTANEOUS ESTIMATION OF ACEBROPHYLLINE AND ACETYLCYSTEINE
IN CAPSULE DOSAGE FORM . International Journal Of Pharmaceutical And
Phytopharmacological Research (Eijppr) : 1-9.
12. Heyden Y.,Mangelings D. Development And Validation Of An Hplc Method With Post-Column
Derivatisation For Assay Of N-Acetylcysteine In Plasma. Acta Chromatographica 2004 ; 14:
149-164.

18

13. Suarez W.,Vieira H.,Fatibello-Filho O.. Flow Injection Turbidimetric Determination Of

Acetylcysteine In Pharmaceutical Formulations Using Silver Nitrate As Precipitant Reagent.
Journal Of . Brazillian Chemical . Society 2007; 18(5): 1028-1033
14. Amir W,Mohammad Y, Abdul N. Flow-Injection Determination Of Cysteine, N-Acetyl Cysteine
And Glutathione In Pharmaceuticals Via Potassium Ferricyanide-Fe(III) Spectrophotometric
System . Journal Of Chemical Research 2010; 26(6): 893898
15. Gabard B. Endogenous Plasma N-Acetylcysteine And Single Dose Oral Bioavailability From
Two Different Formulations As Determined By A New Analytical Metho. Biopharmaceutics &
Drug Disposition 1991; 12: 343-353 .
16. Ercal n.,oztezcan s.,matthews r.. High-performance liquid chromatography assay for nacetylcysteine in biological samples following derivatization with n-( 1-pyrenyl)maleimide .
Journal of chromatography b: biomedicala pplications 1996; 685: 329-334 .
17. Baeyens W., Van Der Weken G., Lin Ling B.. Hplc Determination Of N-Acetylcysteine In
Pharmaceutical Preparations After Pre-Column Derivatization With Thiolyter MB Using
Fluorescence Detection . Analytical Letters 2006; 21(5): 741-757
18. Taha E.,Hassan N., Abdel Aal F., Fattah L.. Kinetic Spectrophotometic Determination of
Acetylcysteine and Carbocisteine in Bulk Powder and in Drug Formulations. ScienceAsia 3
2008; 34: 107-113.
19. Sana S., Rajani A., Sumedha N.. Development and Validation of RP-HPLC Method for the
Estimation of N- Acetylcysteine in Wet Cough Syrup. International Journal of Drug
Development & Research 2012; 4(2): 285-293.
20. Toussaint B.,Pitti C.,Ceccato A.. Quantitative analysis of N-acetylcysteine and its pharmacopeial
impurities in a pharmaceutical formulation by liquid chromatographyUV detectionmass
spectrometry. Journal of Chromatography A 2000; 896: 191199.
21. Siddiqui M., Wabaidur S.,Alothman Z.. Iodate Oxidation Of N-Acetyl L-Cysteine: Application
In Drug Determination And Char- Acterization Of Its Oxidation And Degradation Product By
Mass Spectrometry . Journal Of The Chilean Chemical Society 2014; 59(N1): 2303-2307.
22. Kukoc-Modun L., Radi N.. Spectrophotometric Determination of N-Acetyl-L-Cysteine and N(2-Mercaptopropionyl)-Glycine in Pharmaceutical Preparations. International Journal of
Analytical Chemistry 2011; 2011: 1-6.
23. Longo A.,Toro M.. Determination of N-acetylcysteine in human plasma by gas chromatography
mass spectrometry. Journal of Chromatography B:Biomedical applications 1991; 562(1-2):
639645.
24. Ghannam S.,Brashy A.. Fluorimetric determination of some thiol compounds in their dosage
forms. Il Farmaco 2002; 57(8): 625629
25. U.S. Pharmacopoeia XXIX. Acetylcysteine Monograph; 2005, pp. 51

19

26. U.S. Pharmacopoeia XXIX. Acetylcysteine solution Monograph; 2005, pp. 51

27. European Pharmacopoeia, 5th ed. N-Acetylcysteine Monograph; 2007, pp. 915917.
28. USP Medicines Compendium. https://mc.usp.org/monographs/all/A (accessed 15 october 2014).
29. PUBCHEM. Acetyl cysteine http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?
cid=12035&loc=ec_rcs (accessed 15th October 2014).
30. Ourique A.,Coradini K.. A LC UV method to assay N- acetyl cysteine without derrivitization
:analysis of pharmaceutical products . Analytical methods 2013; 5: 3321-3327.
31. Farquhar J.,Finlay G.. A reversed phase high performance liquid chromatographic assay for the
determination of N-acetyl cysteine in aqueous formulations. Journal of pharmaceutical and
biomedical analysis 1985; 3(3): 279-285.
32. Fohl A.,Johnson C.. Stability of extemporaneously prepared acetyl cysteine 1% and 10
%solutions for treatment of meconium ileus . American journal of health systems pharmacy
2011; 68: 69-72.
33. The phung to,Ellis A.,Ching M.et al . Stability of a formulated N-acetyl cysteine capsule for
prevention of contrast induced nephropathy . Journal of pharmacy practice and research 2008.
34. Siden R.,Johnson C.. Stability of a flavoured formulation of acetyl cysteine for oral
administration . American journal of health systems pharmacy 2008; 65(Mar 15): 558-561
35. Seo E.. Stability of N-acetyl cysteine in peritoneal dialysis solution . Peritoneal dialysis
international 2010; 30: 105-111.
36. Dribben W.,Porto S.. Stability and microbiology of inhalant N-acetyl cysteine used as an
intravenous solution for the treatment of acetaminophen poisoning . Annals of emerging
medicine 2003; 42(1): 9-13.

20

21

22