SIMULTANEOUS ESTIMATION OF LEVOCETRIZINE DIHYDROCHLORIDE AND AMBROXOL HYDROCHLORIDE IN SYRUP DOSAGE FORM BY USING UV SPECTROPHOTOMETRIC METHOD
S. S. Pekamwar, Madhav Shelke
ABSTRACT: A simple, economical, rapid, accurate and precise UV Spectrophotometric method was developed for simultaneous estimation of Levocetrizine dihydrochloride and Ambroxol hydrochloride in bulk and in Syrup formulation. Determination of Levocetrizine dihydrochloride (LCTZ) and Ambroxol hydrochloride (AMB) in bulk and in syrup was conducted by using simultaneous equation method based on measurement of absorbance at 246 nm and 231 nm, λmax of LCTZ and AMB, respectively. The solvent used for proposed method was methanol: water (50:50). The linearity for LCTZ and AMB was in the range of 5-35 µg/ml and 5-50 µg/ml, respectively. The developed method was validated as per ICH guidelines. The % recovery was found to be in the range of 99.52- 99.82% and 100.37- 100.72% for LCTZ and AMB, respectively. Thus the proposed method was simple, rapid, reproducible and accurate and can be successfully applied for simultaneous estimation of Levocetrizine dihydrochloride and Ambroxol hydrochloride in bulk and in Syrup formulation.
KEYWORDS: Levocetrizine dihydrochloride, Ambroxol hydrochloride, Simultaneous Equation Method, UV Spectrophotometry, Syrup, Validation.
INTRODUCTION: Levocetirizine is a third generation non-sedative antihistamine. Chemically, levocetirizine is the active enantiomer of cetirizine. It is the L-enantiomer of the cetirizine racemate. Levocetirizine works by blocking histamine receptors. Levocetrizine is chemically, (R)-2-[2-[4-[(4- chlorophenyl)phenylmethyl] piperazin-1-yl] ethoxy] acetic acid dihydrochloride.
Chemical Structure of Levocetrizine dihydrochloride:
Ambroxol is an active N-desmethyl metabolite of mucolytic bromhexine. Ambroxol is used as a bronchosecretolytic and expectorant drug. Ambroxol has been shown to increase the quantity and decrease the viscosity of respiratory tract secretions. It enhances pulmonary surfactant production (surfactant activator) and stimulates ciliary motility. These actions result in improved mucus flow and transport (mucociliary clearance). Enhancement of fluid secretion and mucociliary clearance facilitates expectoration and eases cough. Ambroxol is chemically, trans-4-[(2-Amino-3,5-dibromobenzyl)amino] cyclohexanol hydrochloride.
Chemical Structure of Ambroxol hydrochloride:
Literature survey revealed that various analytical methods involving UV Spectrophotometry, HPLC and HPTLC have been reported for LCTZ and AMB alone and in Combination with other drugs. However there is no method developed for simultaneous estimation of Levocetrizine dihydrochloride and Ambroxol hydrochloride in liquid dosage form (Syrup). The objective of present work was to develop simple, rapid, economical accurate and precise analytical method for simultaneous estimation of Levocetrizine dihydrochloride and Ambroxol hydrochloride in bulk and in liquid dosage form (Syrup). The developed method was validated in accordance with ICH guidelines and successfully employed for the assay of LCTZ and AMB in combined liquid dosage form (Syrup).
MATERIALS AND METHODS
Chemicals and Reagents: Analytically, pure LCTZ and AMB were provided as gift sample by Bidwai Chemicals Pvt. Ltd. Nanded and Glenmark Pharmaceuticals, Mumbai, Maharashtra, respectively. Syrup of LCTZ and AMB, Mzic D manufactured by Monad Pharmaceuticals, were purchased from local pharmacy. Methanol AR grade (Molychem) and Distilled water was used throughout the experimental work.
Instruments: A UV-Visible Spectrophotometer: Shimadzu, Model: UV-1800 with 1 cm matched quartz cell and UV Probe 2.33 software was used. Calibrated analytical balance Anamed was used for weighing purpose. Digital Ultrasonic Cleaner (Sonicator): HMG India and Vaccum Pump Assembly: Millipore was used for the experimental work.
Preparation Standard Stock Solution:
Ambroxol hydrochloridre standard stock solution:
Accurately weighed reference standard of AMB (10mg) was transferred to 100ml volumetric flask and dissolved in 50 ml solvent (Water, methanol 1:1) and sonicate for 5 min. then volumes was made up to the mark with same solvent; to obtain standard stock solution (100µg/ml) of drug. For the preparation of working standards, suitable aliquots of stock solution were pipette out and volumes were made up to the mark with solvent (Water, methanol 1:1); so as to get required concentrations.
Levocetrizine dihydrochloride standard stock solution:
Accurately weighed reference standard of LCTZ (10mg) was transferred to 100ml volumetric flask and dissolved in 50 ml solvent (Water, methanol 1:1) and sonicated for 5 min. then volumes was made up to the mark with same solvent; to obtain standard stock solution (100µg/ml) of drug. For the preparation of working standards, suitable aliquots of stock solution were pipette out and volumes were made up to the mark with solvent (Water, methanol 1:1); so as to get required concentrations.
Selection of analytical wavelength:
Sample solutions were scanned over the wavelength range of 200 nm to 400 nm, λmax for AMB and LCTZ were found at 231 nm and 246 nm. Representative absorption spectra of AMB and LCTZ are shown in fig. 1 & 2.
Fig. 1 : Absorption Spectra of LCTZ
Fig. 2 : Absorption spectra of AMB
Preparation of calibration curve of AMB and LCTZ
By appropriate dilution of standard stock solution, different dilutions were prepared ranging from 5µg/ml to 35µg/ml for LCTZ and 5 µg/ml to 50 µg/ml for AMB. Absorbance of all the dilutions was plotted against the respective concentrations to obtain the calibration curves. The absorption overlain spectrum of AMB and LCTZ is shown in fig. 3
Fig. 3 : Overlay Spectra of LCTZ & AMB
Fig. 4 : Calibration Curve of LCTZ
Fig. 5 : Calibration Curve of AMB
Analysis of Marketed Syrup Formulation:
Ten syrup bottles were weighed accurately, average weight was determined and poured in calibrated measuring cylinder to check extractable volume. As per label claim syrup contain LCTZ 5mg/5ml and AMB 30mg/5ml, so 5ml syrup was pipette out in 100 ml volumetric flask and dissolved in water: methanol (1:1) solution, sonicated for 10 min and filtered. Then different concentrations of syrup sample were prepared by serial dilution method and used for analysis. Prepared syrup sample was analysed on UV spectrophotometer using Simultaneous Equation Method.
Method Validation
The method validation parameters linearity, precision, accuracy, repeatability, limit of detection and limit of quantitation were checked as per ICH guidelines.
Precision: Precision of the method was verified by using syrup stock solution, the repeatability indicates the performance of the UV instrument. Interday and intraday precision was determined by repeating the assay three times in same day for intraday precision and on different day for interday precision studies.
Accuracy (Recovery Study): To check the accuracy of proposed method, recovery studies were carried out at 80%, 100% and 120% of the test conc. As per ICH guidelines.
To perform recovery study at 100%, 5ml syrup pipette out in volumetric flask and to this standard 30mg AMB and 5mg LCTZ was added. The volume as made up to the mark with water: methanol (1:1) solution and further dilution with same solvent to obtain sample solution Similarly, for 80% recovery study, 26mg AMB and 4mg LCTZ were added, and for 120% recovery study, 36mg AMB and 6mg LCTZ were added respectively.
RESULTS AND DISCUSSION
Precision
Intraday Precision:
Sr. No.
Interval of
Concentration (µg/ml)
% Recovery
Time
AMB
LCTZ
I
30.10
4.96
100.34
99.35
II
Intra-day
29.95
4.95
99.84
99.28
III
30.20
4.97
100.68
99.54
Table 1 : Intra-day Precision data of marketed formulation
Name of the drug
Mean*
SD
%RSD
AMB
100.28
0.4225
0.4213
LCTZ
99.40
0.1345
0.1353
* Indicates average of three determinations
Table No. 2 : Statistical validation of intra-day precision data
Inter-day Precision:
Sr. No.
Interval of
Concentration (µg/ml)
% Recovery
Time
AMB
LCTZ
AMB
LCTZ
I
30.14
4.98
100.45
99.79
II
Inter-day
30.09
4.98
100.32
99.72
III
29.96
4.98
99.87
99.63
Table 3 : Inter-day Precision Data
Name of the drug
Mean*
SD
%RSD
AMB
100.21
0.3043
0.3036
LCTZ
99.71
0.0802
0.0804
* Indicates average of three determinations
Table 4 : Statistical validation of inter-day precision data
Accuracy (Recovery study):
Level of Recovery
Amount present
(mg/5ml)
Added concentration (mg)
Amount recovered
(mg)
% Recovery
AMB
LCTZ
AMB
LCTZ
AMB
LCTZ
AMB
LCTZ
30
5
24
4
54.31
8.97
100.58
99.74
80%
30
5
24
4
54.20
8.95
100.38
99.52
30
5
24
4
54.38
8.98
100.72
99.82
30
5
30
5
60.28
9.96
100.47
99.68
100%
30
5
30
5
60.32
9.97
100.54
99.70
30
5
30
5
60.22
9.95
100.38
99.54
30
5
36
6
66.31
10.96
100.48
99.69
120%
30
5
36
6
66.36
10.96
100.56
99.72
30
5
36
6
66.24
10.95
100.37
99.58
Table 5 : Recovery study data
Level of
recovery
% Mean recovery
S.D.
% R.S.D.
AMB
LCTZ
AMB
LCTZ
AMB
LCTZ
80%
100.56
99.70
0.1708
0.1553
0.1698
0.1557
100%
100.46
99.64
0.0802
0.0871
0.0798
0.0874
120%
100.47
99.67
0.0953
0.0737
0.0948
0.0951
Table 6 : Statistical validation of recovery study data
Limit of Detection and Limit of Quantitation:
Name of the drug
LOD µg/ml
LOQ µg/ml
AMB
3.1645
9.5896
LCTZ
3.0891
9.3611
Table 7 : LOD & LOQ
CONCLUSION:
The method used is simple and rapid and does not involve the use of complex instrument, low value of Standard deviation showed that the method is precise and high percentage of recovery of as shown in table shows that the method is accurate. Thus from the results it can be concluded that above developed UV method is suitable for estimation of AMB and LCTZ in syrup formulation. The estimated method is sensitive, precise, accurate, specific and economic. Hence, the method can be used successfully for quality control and routine analysis of finished pharmaceutical dosage form.
REFERENCES:
Indian Pharmacopoeia 2007, The Indian Pharmacopoeia Commission, Government of India, Ministry of Health & Family Welfare, Ghaziabad, Volume 2, 83 , 672.
Makarand Avhad, Dr. C.G. Bonde, development and validation of simultaneous uvspectrophotometric Method for the determination of levofloxacin and ambroxol in tablets, International Journal of ChemTech Research, 2009, 1, 4, 873-888
P.A. Patel, M.N. Dole, P.S. Shedpure, S.D. Sawant, Spectrophotometric Simultaneous Estimation of Salbutamol And Ambroxol In Bulk And Formulation, Asian Journal of Pharmaceutical and Clinical Research, 4, 2011, 42-45
Ilangovan Ponnilavarasan, Chebrolu.Sunil Narendra Kumar, and P.Asha, Simultaneous Estimation of Ambroxol Hydrochloride and Loratadine in Tablet Dosage form by Using Uv Spectrophotometric Method, 2.2011, 338-345
Jawed Akhtar, Monica Padaria, Development & Validation of UV Spectrophotometric Methods for Simultaneous Determination of Doxofylline & Ambroxol Hydrochloride in Bulk & their Combined Dosage Form, Indo Global Jr. of Ph. Sciences, 2013, 3(1), 90-95
Ritu Kimbahune, Sunil K, Prachi Kabra, Kuldeep Delvadiya, Sanjay Surani, Spectrophotometric Simultaneous Analysis of Ambroxol Hydrochloride, Guaifenesin and Terbutaline Sulphate In Liquid Dosage Form (Syrup), International Journal of Pharmaceutical Sciences Review And Research, 8, 2, 2011, 24-28
Jyostna S.Patil, Chandan R. Rane, Radhika D.Ingale, Simultaneous Spectrophotometric Methods for Estimation of Levocetirizine and Ambroxol in Pharmaceuticalv Dosage Form, Journal of Chemical, Biological and Physical Sciences, 2013, 3, 2, 1115-1122
S Lakshmana Prabu, AA Shirwaikar, Annie Shirwaikar, C Dinesh Kumar, G Aravind Kumar, Simultaneous UV spectrophotometric estimation of ambroxol hydrochloride and levocetirizine dihydrochloride, Indian J Pharm Sci, 70(2), 2008, 236–238.
Kishore Konam, Pottabathula Hari Prasad, Punuri Samuel Lukaraju, Sirajudeen. M.A, Determination of montelukast sodium and levocetrizine hydrochloride by using HPTLC method, Elixir International Journal, 38, 2011, 4276-4278
Madhura V Dhoka, Shakuntala S. Chopade, Method Development & Comparative Statistical Evaluation of HPLC & HPTLC Method for Simultaneous Estimation of Cefodrixil Monohydrate & Ambroxol Hydrochloride, Indo Global Journal of Pharmaceutical Sciences, 2(2), 2012, 203-212
Prince Francis Moses, Prathap Singh, Alagar Raja, David Banji , Analytical Method Development and Validation for Simultaneous Estimation of Ambroxol and Desloratadine in its Pharmaceutical Dosage Form By Rp-Hplc, World Journal Of Pharmacy And Pharmaceutical Sciences, 2(6), 2013, 6246-6262
Rathore AS, Sathiyanarayanan L, Mahadik KR, Development of Validated HPLC and HPTLC Methods for Simultaneous Determination of Levocetirizine Dihydrochloride and Montelukast Sodium in Bulk Drug and Pharmaceutical Dosage Form, Pharm Anal Acta, 1(1), 2010, 1-6
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