A Review: Current Analytical Methods for Determination of
Atopine Sulphate in Pharmaceutical dosage forms
Hiral M. Pokiya1*, Priyanka S. Malani1, Bhumi K. Patel1, Dr. Hasumati A. Raj1, Dr. Vinit C. Jain1
1
Shree Dhanvantary Pharmacy College,Kim, Surat, Gujarat
Address For correspondence:
Department of Quality Assurance
Shree Dhanvantary Pharmacy College
At : Kim, Taluka : Olpad, Dist: Surat, Pin code : 394110
Mobile No: 8866109499
*Email: pokiyahiral@yahoo.in
Number of figures: 1
Number of tables: 3
ABSTRACT:
Atropine is an anticholinergic or parasympatholytic drug. It inhibits the muscarinic actions of
acetylcholine. Which is available in the different pharmaceutical dosage forms through
various routes of administration, such as oral, ophthalmic and systemic. It is used to produce
mydriasis and cycloplegic effect. This article reviews from different journals in which
developed analytical methods for identification and quantitative determination of Atropine as
a single or combination with other drugs in bulk and pharmaceutical dosage form. The most
commonly used methods for the determination of Atropine presented in this article are
chromatographic methods: HPLC,RP-UPLC,UV spectrophotometric methods were reported.
KEY WORD:-Atropine Sulphate, Chromatographic method, UV spectroscopic method
1) INTRODUCTION:
Atropine Sulphate[Bis[(1R,3R,5S)-8-methyl-8-azabicyclo[3.2.1]oct-3-yl (2RS)-3-hydroxy-2phenylpropanoate]sulphate] is appears as a White Crystalline Powder (Figure 1)(1).Atropine is a
naturally occuring tropen alkaloid and basic in nature. It is obtained from Atropa belladonna
and Solanaceous family(2). It is Very soluble in water, freely soluble in methanol and ethanol(3)
Atropine Sulphate has pKa values is 9.9(4) and melt at Not lower than 187°C(5)
Figure 1: Chemical structures of Atropine
Sulphate(1)
Atropine is an anti-muscarinic agents or parasympatholytic drug. It is act on muscarinic
receptor and inhibit the action of acetylcholine. This results in dilation of the pupil (mydriasis)
and unresponsive to light. Another effect is relaxation of ciliary muscle causes paralysis of
accomodation (cycloplegia) and near vision is impaired(6)
2) METHODS FOR THE DETERMINATION OF DEXAMETHASONE SODIUM
PHOSPHATE
a) Compendial Method:
Atropine Sulphate is official in Indian pharmacopoeia, British
Pharmacopoeia,
United
State
Pharmacopoeia
and
European Pharmacopoeia and estimation is done by potentiometric
titration(table-1)
Table-1 Official compendial methods for assay of Atropine Sulphate
INDIAN PHARMACOPOEIA(1)
1)
Method
Potentiometric titration
Procedure Weigh drug and dissolve in anhydrous glacial acetic acid.Titrate
with 0.1 M perchloric acid.
Factor :- 1 ml of 0.1 M perchloric acid is equivalent to 0.06768 g
of Atropine sulphate.
BRITISH PHARMACOPOEIA(3)
2)
Method
3)
Potentiometric titration
Procedure Weigh drug and dissolve in anhydrous glacial acetic acid.Titrate
with 0.1 M perchloric acid.
Factor :- 1 ml of 0.1 M perchloric acid is equivalent to 0.06768 g
of Atropine sulphate
UNITED STATES PHARMACOPOEIA(5)
Method
Potentiometric titration
Procedure
Weigh drug and dissolve in anhydrous glacial acetic acid.Titrate
with 0.1 M perchloric acid.
Factor :- 1 ml of 0.1 M perchloric acid is equivalent to 0.06768
g of Atropine sulphate.
EUROPEAN PHARMACOPOEIA(7)
4)
Method
Potentiometric titration
Procedure
Weigh drug and dissolve in anhydrous glacial acetic acid.Titrate
with 0.1 M perchloric acid.
Factor :- 1 ml of 0.1 M perchloric acid is equivalent to 0.06768
g of Atropine sulphate.
b) Spectrophotometric Method:
Table-2 Developed spectroscopic method for Atropine Sulphate
Sr. Title
No.
1.
The Use of the First-derivative
Curves of Absorption Spectra
in Quantitative Analysis
2.
Spectrophotometric
Determination of Atropine
Sulfate in Eye Drops using
Bromthymol Blue
3.
Colorimetric Determination of
Atropine,
Homatropine,
Scopolamine,
and
their
Derivatives by the Ferric
Hydroxamate Method
Abstract
Ref.
No.
The first-derivative curves of Uv-Visible spectra 8
(ΔE/δλ ) are useful in quantitative analysis. A
base-line technique and its algebraic version are
reported for (i) the correction of quadratic
irrelevant absorption, and (ii) the determination of
a substance in the presence of another with
overlapping spectra. The method is illustrated by
the determination of atropine sulphate in the
presence of phenyl-mercury(ii) nitrate. The results
obtained suggest that the method warrants careful
study over a wide field of applications.
Content of atropine in eye drops of atropine sulfate 9
were
determined
colorimetrically
and
spectrophotometricaly using methods of ion pairs
with bromtimolblue. The described method is
more convenient for routine analysis then non
aqueous
titration
described
in
various
Pharmacopoeias.
The purpose of this study was to develop a 10
procedure for the quantitative analysis of the salts
of the solanaceous alkaloids in the presence of
their hydrolysis products. The salts taken were
atropine sulfate, homatropine hydrobromide,
homatropine
methyl-bromide,
scopolamine
hydrobromide, and scopolamine methyl-bromide.
The ferric hydroxamate procedure was used. The
conditions necessary for a reproducible reaction
were established; a procedure of sufficient
sensitivity was developed which will allow unit
dose analysis; and an analysis was developed
which will permit the salts of the solanaceous
alkaloids to be quantitatively analyzed in the
presence of their degradation products.
c) Chromatographic method
Table-3 Developed Chromatographic method for Atropine Sulphate
Sr.No. Title
Mobile phase
1.
Water: Acetonitrile:
Tetrahydrofuran
(67:30:3,
v/v/v)
containing 1 mM
perchloric acid
Methanol:5 mmol 264 nm
Potassium
dihydrogen
Phosphate
buffer
(50: 50, v/v)
2.
3.
4.
5.
6.
7.
8.
High Performance Liquid Chromatographic
Determination of Atropine and Atropine
like
Alkaloids
in
Pharmaceutical
Preparations
with
Indirect
Conductometric Detection
RP-HPLC Method Development for
Estimation of Atropine Sulphate in Bulk
Impurity Profiling of Atropine Sulfate by
Microemulsion
Electrokinetic
Chromatography
Development and Validation of Novel RP- Phosphate buffer :
UPLC Method for Estimation of Atropine Acetonitrile (87:13,
Sulphate in Pharmaceutical Dosage Form
v/v)pH was adjusted
to 3.5 by ortho
phosphoric acid
Simultaneously Determination of Atropine CH3
CN-0.1%
Sulphate, Diphenhydramine Hydrochloride, H3PO4
Capsaicin and Dihydrocapsaicin in Painrelieving Plaster for Arthritis by RP-HPLC
LC Determination of Atropine Sulfate and Methanol : Water :
Scopolamine
Hydrobromide
in Formic
acid
Pharmaceuticals
(165:35:1) and pH
adjusted 8.3 with
triethylamine
Determination of Atropine and Obidoxime
in Automatic Injection Devices Used as
Antidotes
Against
Nerve
Agent
Intoxication
Determination of Atropine Sulphate and acetonitrile-diluted
Benzalkonium Chloride in Eye Drops by acetic acid (80:20,
HPLC
v/v)
Wavele
ngth
Ref.No.
11
12
13
210 nm
14
280 nm
15
230nm
16
17
-
260 nm
18
3) DISCUSSION
The presented review highlights on various analytical methods published on Atropine Sulphate
and combination with other drug. HPLC-UV methods were found to be most sensitive for
Atropine Sulphate. Various chromatographic conditions are presented in under Table 3. The
HPLC method could be automated; there are different column fillings; different solvents with
different polarity as mobile phases and different detection modes. The faster time, high
sensitivity, specificity and better separation efficiency enable HPLC to be used frequently for the
simultaneous qualitative and quantitative determination of Atropine Sulphate in the comparison
with the other methods. There is no doubt on the fact that these chromatographic methods are
rapid and far more economical .In this way various analytical methods for the estimation of these
Atropine Sulphate in bulk or in various matrixes like plasma, alone or in combination with other
drugs is discussed. The presented information is useful for the researchers especially those
involved in the formulation development and quality control of Atropine Sulphate and
combination with other drug.
4) REFERENCES:
1) Indian Pharmacopoeia, The Indian Pharmacopoeia Commission Ghaziabad, Govt. of India
Ministry of Health and Family Welfare,vol.II:852-853,2010.
2) Rang HP, Dale MM, Ritter JM, Flower RJ, “Rang and Dale’s Pharmacology”,6 th edition,
Edinburgh:Churchill Livingstone,153-154, 2008.
3) British Pharmacopoeia, The Department of Health, London: The
193-195, 2010.
Stationary Office,vol.I:
4) Moffat AC, et al., “Clarke’s Isolation and Identification of Drugs”, 2nd edition, The
Pharmaceutical Press ,London - GB, 363-364,1986.
5) The United States Pharmacopoeia (USP 32 NF 27) Asian Edition, Rockville, MD:USP
Convention Inc.,vol.II:1604, 2009.
6) Tripathi KD, “Essentials of Medical
Medical Publishers (P) Ltd;106-110, 2008.
Pharmacology”,6 th edition, Jaypee Brothers
7) The European Pharmacopoeia, published by the European Directorate for the Quality of
Medicines & Health Care (EDQM),7th edition,vol.II:1429-1430,2011.
8) Wahbi AM and Ebel S, “The Use of the First-Derivative Curves of Absorption Spectra in
Quantitative Analysis”, Analytica Chimica Acta, 1974;70(1):57-63.
9) Polomik M, Sober M , Pleho A , Nikolin B, “Spectrophotometric Determination of Atropine
Sulfate in Eye Drops using Bromthymol Blue”, Medicinski arhiv., 1993;47(1-2):25-27.
10) Feldman joseph A and robb Bruce J, “Colorimetric Determination of Atropine, Homatropine,
Scopolamine, and their Derivatives by the Ferric Hydroxamate Method”, Journal of
Pharmaceutical sciences, 1970;59(11):1646–1647.
11) Oi – Wah Lau and Chuen-Shing Mok, “High Performance Liquid Chromatographic
Determination of Atropine and Atropine like Alkaloids in Pharmaceutical Preparations with
Indirect Conductometric Detection”, Journal of Chromatography A,1997;766(1-2):270-276.
12) Raghuwanshi AS and Jain UK, “RP-HPLC Method Development for Estimation of Atropine
Sulphate in Bulk”, Oriental Journal of Chemistry, 2009;25(3):621-624.
13) Bitar Yaser and Holzgrabe Ulrike, “Impurity Profiling of Atropine Sulfate by Microemulsion
Electrokinetic Chromatography”, Journal of Pharmaceutical and Biomedical Analysis,
2007;44(3):623-633.
14) Arvadiya AC and Dahivelker PP, “Development and Validation of Novel RP-UPLC Method
for Estimation of Atropine Sulphate in Pharmaceutical Dosage Form”, Chemical Industry &
Chemical Engineering Quarterly,2013;19(3):333-337.
15) Jia X, et al., “Simultaneously Determination of Atropine Sulphate, Diphenhydramine
Hydrochloride, Capsaicin and Dihydrocapsaicin in Pain-relieving Plaster for Arthritis by RPHPLC”, Zhongguo Zhong Yao Za Zhi, 2010;35(21):2838-2841.
16) Kartal M, et al., “LC Determination of Atropine Sulfate and Scopolamine Hydrobromide in
Pharmaceuticals”, Journal of Pharmaceutical and Biomedical Analysis,2001;25(3-4):399406.
17) Juhani Pohjola and Marco Harpf, “Determination of Atropine and Obidoxime in Automatic
Injection Devices Used as Antidotes Against Nerve Agent Intoxication”, Journal of
Chromatography A, 1994;686(2):350-354.
18) Pinzauti S, et al., “Determination of Atropine Sulphate and Benzalkonium Chloride in Eye
Drops by HPLC”, International Journal of Pharmaceutics,1993;93(1-3):239-243.