Int J Pharm Pharm Sci, Vol 6, Issue 9, 394-397Original Article

A STABILITY INDICATING UV SPECROPHOTOMETRIC METHOD FOR DETERMINATION OF METOCLOPRAMIDE HYRDROCHLORIDE

U A DEOKATE, ANJALI MACCHINDRA GORDE*

Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India 431005
Email: gordeanjali@gmail.com

Received: 07 Jul 2014 Revised and Accepted: 27 Aug 2014


ABSTRACT

A stability indicating method has been developed for specific determination of Metoclopramide HCl in bulk by UV spectrophotometry in presence of its degradation products. The method is simple, accurate, precise, and robust. Linearity range for the method is 10-50µg/ml at detection wavelength of 272 nm. The LOD and LOQ values were found to be 3.26µg/ml and 9.89µg/ml respectively.

Keywords: Metoclopramide hydrochloride, Stability indicating, UV Bulk, Forced degradation.


INTRODUCTION

Metoclopramide HCl, 4-amino-5-chloro-N-[2-(diethylamino)ethyl]-2-methoxybenzamide hydrochloride, is an antiemetic drug. It inhibits gastric smooth muscle relaxation produced by dopamine and therefore increases cholinergic response of gastrointestinal tract. [1,2]

Knowledge of the stability of the molecule helps in selecting proper formulation and package as well as providing proper storage conditions and shelf life, which is essential for regulatory documentation. Forced degradation is a process that involves degradation of drug products and drug substances at conditions more severe than accelerated conditions and thus generates degradation products that can be studied to determine the stability of the molecule [3]. The presently available literature reveals few analytical methods for determination of Metoclopramide HCl by HPLC [4,5], UV spectrophotometry [6,7], LC-MS [8], in plasma [9].

The objective of this work was to develop a validated Stability indicating assay method for the determination of Metoclopramide hydrochloride in presence of its degradation products generated by subjecting the drug to forced degradation conditions under acid, alkali, thermal and oxidative stress as per the guidelines[10] and to establish the inherent stability of Metoclopramide hydrochloride.

MATERIALS AND METHODS

Metoclopramide hydrochloride was supplied by IPCA Laboratories, Aurangabad as the gift sample. Its identity and purity were confirmed by recording the FTIR spectra. Solvent used was Deionized water from the water treatment plant (SG) of the Institute. The reagents used were Hydrochloric acid (Qualigens), Sodium hydroxide & Hydrogen Peroxide 50% solution.

Apparatus

UV Spectrophotometer- Shimadzu Japan UV 1700, FT-IR- Thermo electron co.(IR 200) Prestige 21 and analytical balance (Schimadzu AUX 220).

Method

Spectrophotometric conditions

Spectrophotometric analysis was carried out at ambient temperature. Solvent used was deionized water; standard solution of 50µg/ml was analyzed at 272 nm.

Preparation of stock and standard solution

Accurately weighed 100mg of standard Metoclopramide HCl was transferred to 100 ml volumetric flask to make a stock solution of 1000µg/ml. Suitable aliquots were taken into 100 ml volumetric flask to make standard solutions in the range of 10-50 µg/ml.

Preparation of calibration curve

The calibration curve was prepared in the concentration range 10-50μg/ml by analyzing each solution in triplicate and plotting the concentration (μg/ml) against absorbance. The correlation coefficient and equation of the line were determined. The spectrum of fresh drug solution (50µg/ml) is shown in fig1 and the calibration curve in fig 2. The data is shown in table 2.

Forced degradation studies

The forced degradation of MTD was done in each of the stress condition at a concentration of 1 mg/ml. The degradation was confirmed in each case by recording the changes in the ultraviolet spectra of each stressed sample comparing it with that of fresh drug solution.

A. Acidic stress

100 mg of MTD was refluxed in 1 N Hydrochloric acid for 4 hours in a water bath at 70oC.

B. Alkaline stress

100 mg of MTD was refluxed in 1 N sodium hydroxide for 4 hours in a water bath at 70oC.

C. Thermal stress

100 mg of MTD was kept in solid state at 70oC for 4 hours in an oven equipped with temperature control probe.

D. Oxidative stress

100 mg of MTD was dissolved in 3% and 6% Hydrogen peroxide separately and kept at room temperature for 24 hours in amber colored stoppered vials.

RESULTS AND DISCUSSION

Recording of spectrum of standard solution

The spectrum of standard drug was recorded with 50µg/ml solution, and scanned in the range of 400-200 nm. 272 nm was found to be the suitable wavelength for analysis.

Degradation behavior of MTD

The amount (percent) of MTD remaining undegraded under each stress condition was determined from the absorbance of the fresh drug sample relative to that of stressed drug sample. The results are shown in table 7.

Method validation

A. Accuracy

Accuracy of the method was evaluated by spiking the drug at three concentration levels (8, 10 & 12µg/ml) to the original 10 µg/ml solution. The percent recovery of the added drug was calculated from the linearity plots. The results are shown in table 1.

B. Linearity and range

The linearity of the method was established by preparing a calibration curve in water in the range 10-50μg/ml. Triplicates of each of the solution were analyzed and calibration curve recorded. The mean (n=3) absorbance was plotted against concentration (μg/ml). The correlation coefficient and equation of line were determined. The results are shown in fig2 and table 2.

C. Precision

The Interday and intraday precision was determined by calculation of the % RSD values on triplicates of each concentration. The mean (n=3) absorbance of each concentration was compared with that of the second run on the same day (intraday) and with that on the next day (interday) and the percent relative deviation calculated. The results are shown in table 3.

D. Specificity

The Specificity of the method was established through the determination of the drug in the presence of its degradation products with high degree of precision. The spectrum homogeneity was confirmed by analyzing the ratio chromatograms at the wavelengths 271 nm and 273 nm.

E. Ruggedness [16]

Mean absorbance (n=3) was measured for the 10μg/ml solution analyzed by two different analysts on different days and the percent relative deviation between two trials was calculated. The results are shown in table 4.

Fig. 1: Spectrum of fresh standard solution of Metoclopramide HCl (50µg/ml)


Fig. 2: Calibration curve of Std MTD

F. Robustness

Deliberate changes in the detection wavelength were made. Three replicates of each deviation were analyzed and the %RSD between the mean (n=3) absorbance and that obtained under optimized spectrophotometric conditions were determined. The results are shown in table 5.

G. Limit of detection and limit of quantitation

The limit of detection and limit of Quantitation was calculated based on standard deviation (σ) and the slope (S) of the calibration plot, using the formulae LOD= 3.3σ/s and LOQ=10σ/S as defined by ICH. The LOD was found to be 3.26µg/ml and LOQ 9.89µg/ml.

Fig. 3: Spectrum of Acid degraded MTD


Fig. 4: Spectrum of base degraded MTD


Fig. 5: Spectrum of oxidatively (0.3%) degraded MTD


Fig. 6: Spectrum of Oxidatively (0.6%) degraded MTD


Fig. 7: Spectrum of thermally Degraded MTD

H. System suitability

The system suitability parameters Linearity and range, accuracy, precision and specificity were determined and are shown in table 6.

Table 1: Results of recovery studies

Preanalyzed

Sample Solution

(µg/ml)

Level of

Addition

Drug recovered

(µg/ml)

% Recovery %RSD
MTD10 80 17.92 99.55 0.435
100 20.02 100.1 0.698
120 22.03 100.13 0.333
Average %RSD 0.488

Table 2: Linearity Study of MTD at 272 nm

S.

No.

Conc. Absorbance

Mean±S. D.

%RSD
1 10 0.498±0.0032 0.642
2 20 0.883±0.001 0.113
3 30 1.269±0.0015 0.118
4 40 1.648±0.0005 0.030
5 50 2.045±0.0005 0.024

Table 3: Results of Intra-day and Inter-day precision studies

Drug Intra-day Precision  Inter-day Precision
Conc. Mean±S. D. %RSD  Mean±S. D. %RSD
MTD 10 9.99±0.036 0.360  10.005±0.028 0.279
20 19.91±0.035 0.175  19.98±0.077 0.385
30 30.02±0.015 0.050  29.97±0.063 0.210
40 40.07±0.056 0.139  39.96±0.148 0.370
50 49.70±0.045 0.092  49.84±0.197 0.395
 Average %RSD 0.163  Average %RSD 0.327

Table 4: Results of ruggedness studies

Drug Amount Taken
(µg/ml)
Analyst I Analyst II
Amount Found %RSD Amount Found %RSD
MTD 10 9.96±0.025 0.25 9.99±0.0152 0.152
MTD 10  Instrument I  Instrument II
10.01±0.047 0.47 9.80±0.045 0.459

Table 5: Results of Robustness studies

Drug Amount Taken
(µg/ml)
WL-1 (271 nm)
Amount %RSD Found
WL-2 (272 nm)
Amount %RSD Found
WL-3 (273 nm)
Amount %RSD Found
MTD 10 9.88±0.02 0.202 9.96±0.025 0.25 9.99±0.045 0.45

Table 6: System suitability parameters

Validation Parameter Values
Range 10-50µg/ml
Linearity-
Regression equation (y=mx+C) y = 0.038x + 0.110
and Correlation coefficient r2 r² = 0.9998
LOD 3.26µg/ml
LOQ 9.89µg/ml
Recovery (% RSD) 0.488
Intra-Day Precision 0.163
Inter-Day Precision 0.327
Inter-Analyst 0.201
Inter-Instrument 0.464

Table 7: % Drug Degraded under each stress condition

S. No. Stress Condition Absorbance % Drug degraded % Drug Remain
1 Fresh Solution 2.045 0 100
2 Acidic 1.927 5.78 94.22
3 Basic 1.859 9.10 90.90
4 Oxidative (0.3%) 0.290 85.82 14.18
5 Oxidative (0.6%) 0.262 87.19 12.81
6 Thermal 1.639 19.86 80.14

CONCLUSION

A validated stability indicating assay method has been developed for the determination of Metoclopramide hydrochloride in bulk. The results show that the developed method was accurate, precise, simple, economic, fast and specific. Metoclopramide HCl is most prone to degradation under oxidative (6%) stress, followed in order by the stress oxidative (3%), thermal, basic, and acidic.

ABBREVIATIONS

MTD: Metoclopramide HCl.

CONFLICT OF INTERESTS

Declared None

ACKNOWLEDGEMENT

The authors are grateful to IPCA laboratories Ltd. (Aurangabad, India) for kindly providing gift sample of Metoclopramide HCl. Further, we extend our thanks to Dr. R. B. Nawale for valuable suggestions and providing facilities for research work.

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About this article

Title

A STABILITY INDICATING UV SPECROPHOTOMETRIC METHOD FOR DETERMINATION OF METOCLOPRAMIDE HYRDROCHLORIDE

Keywords

Metoclopramide hydrochloride, Stability indicating, UV Bulk, Forced degradation.

Date

01-10-2014

Additional Links

Manuscript Submission

Journal

International Journal of Pharmacy and Pharmaceutical Sciences
Vol 6, Issue 9, 2014 Page: 394-397

Online ISSN

0975-1491

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Authors & Affiliations

Anjali Macchindra Gorde
Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India 431005
India

U A Deokate
Department of Pharmaceutical Chemistry, Government College of Pharmacy, Aurangabad, Maharashtra, India 431005
India


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