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Asian Journal of Pharmacy and Pharmacology 2016; 2(1): 6-9 Zero order and first order derivative Spectrophotometric determination of
Metformin HCL in bulk dosage form
Rajesh Shukla , Sunil Singh *, Jay Ram Patel , Sarita Kar
1Guru Ramdas Khalsa Institute of Science and technology, Barela, Jabalpur, Madhya Pradesh, India
2*Department of Pharmaceutical Chemistry, Oriental College of Pharmacy, Raisen Road Bhopal, Madhya Pradesh, India

Received: 13 January 2016 Revised: 15 February 2016 Accepted: 20 February 2016 Objective: A simple, accurate, sensitive and precise Zero order and First order Ultraviolet spectrophotometric method
has been developed for the determination of Metformine HCL in bulk dosage form. Materials and
methods The
solutions of standard and sample were prepared in methanol. In the UV specrophotometric method, the quantitative determination of the drug was carried for zero order at 234nm and rst order at 246nm, the linearity range was found to respectively. Results: The calibration graphs constructed at their wavelength of
determination were found to be linear for zero and rst derivative spectrophotometric methods. The proposed methods have been extensively validated statistically that included parameters such as linearity, accuracy, precision, LOD, LOQ, recovery and robustness. There was no signicant difference between the performance of the proposed method regarding the mean values and standard deviations.
Conclusion The described methods can be readily utilized for
analysis of pharmaceutical formulation.
Keywords: Method development; validation; derivative spectroscopy; Metformin HCL
combinations in different solvents (Arayne and et al., 2009, Metformin hydrochloride(Dunn and Peters 1995) (Fig.1) (N, N- Amruta et al., 2012). A comparison of the results obtained by dimethylimidodicarbonimidicdiamide hydrochloride) is a biguanide prescribed for the treatment of type II diabetes specrophotometric in the ultraviolet region and obtained by mellitus (Setter et al., 2003) and is the drug of choice in obese HPLC (Chhetri et al., 2014) and other instrumental methods patients (Susan and Jack, 2014). It increases glucose transport of qualitative and quantitative analysis of drugs reveals that across the cell membrane in skeletal muscles (Arayne et al., simple and first order derivative specrophotometric 2009) and it can inhibit the formation of advanced glycosylation determinations can be an economically advantageous end-products (Ruggiero-Lopez et al., 199).
alternative in many cases (Jani et al., 2015). Methods were validated as per the ICH guideline (ICH guideline., 2003).
So, in the present investigation, simple and first order derivative specrophotometric determination of Norfloxacin Figure1: Structure of Metformin HCL
in tablet dosage form is reported.
Pharmaceutical research is developing increasingly complex Materials and methods
molecules and drug formulations, and each novel and highly selective analytical technique is therefore of much potential Analysis carried out on Lab India UV-3200 UV-VIS interest. Literature is enriched with several analytical methods spectrophotometer, a double beam high speed scanning for determination of Metformin HCL in single and in other spectrophotometer (200-800 nm) with a photomultiplier tube detector and having variable spectral bandwidth (0.5- *Address for Corresponding Author: Chemicals and reagents
Oriental College of Pharmacy, Thakral Nagar, Raisen Road, Metformin HCL was received as gratis sample by Bhopal, Madhya Pradesh, India 462021 Aurobindo Pharma Ltd, Hyderabad. All chemicals used E-mail: rssunil29@gmail.com; Phone#: 91-8859120888 were of analytical grade (E. Merck, India).



Asian Journal of Pharmacy and Pharmacology 2016; 2(1): 6-9 Method 1: Development of Zero order spectroscopic method
powder weight equivalent to 1000 mg of metformin was Standard stock solution
accurately taken and transferred to a 50 ml of volumetric To prepare stock solution of Metformin HCL (MTN) (1000 flask. Twenty ml of methanol added to the same and μg/ml), 100 mg of MTN was placed in 100 ml volumetric flask sonicated for 30 min. The flask was shaken, and the volume and dissolved in 75 ml of methanol and the volume was made up was diluted to the mark with the same mixture. The above to the mark with methanol. 10 ml of the solution was diluted up solution was filtered using whatman filter paper no. 1.
to 100 ml with methanol to produce final stock solution of 100 Appropriate volume of the aliquot was transferred to a 50 μg/ml of MTN.
ml volumetric flask and the volume was made up to the mark with methanol. The first derivative spectra were Twenty tablets (METIF-500) were taken, powdered and powder recorded and then measured at 246 nm for MTN. The weight equivalent to 500 mg of metformine HCL was accurately overlain spectra and calibration curve are shown in Figure 3 taken and transferred to a 50 ml of volumetric flask. Twenty ml of methanol added to the same and sonicated for 30 min. The flask was shaken, and the volume was diluted to the mark with the same mixture. The above solution was filtered using whatman filter paper no. 1. Appropriate volume of the aliquot was transferred to a 50 ml volumetric flask and the volume was made up to the mark with methanol. The spectra were recorded and then measured at 234 nm for MTN. The overlain spectra and calibration curve are shown in Figure 2 & 3.
Figure 4 Derivative spectra for Metformin HCL
Different aliquots were pipette out from standard stock solution into a series of 10 ml volumetric flasks and the volume was made up to the mark with methanol to get concentrations of 2, 4, 6, 8, 10, and 12 g/ml of metformin.
Figure 2 Overlain spectra of Metformin HCL
The solutions were scanned on spectrophotometer (Lab India-3200) in the UV range 200-400 nm. The linearity was found 0.999 and range was found 2-12 To the preanalyzed sample solutions (10 Metformin HCL), a known amount of standard stock solution were added at different levels i.e. 80, 100 and 120%. The solutions were reanalyzed by proposed method.
Figure 3 Calibration curve for Metformin HCL
Precision is determined by intra-day and interday precision.
Method 2: Development of first order derivative method
Intra-day precision was determined by analyzing the 6, 8 Standard stock solution
of drug solution for three times in the same To prepare stock solution of MTN (1000 g/ml), day for both proposed methods. Inter-day precision was MTN was placed in 100 ml volumetric flask and dissolved in 75 determined by analyzing the 6, 8 and 10 g/ml ml of methanol and the volume was made up to the mark with solutions daily for over the period of a week for both methanol. 10 ml of the solution was diluted up to 100 ml with proposed methods.
methanol to produce final stock solution of 100 g/ml Repeatability was determined by analyzing 10 Twenty tablets (MET SR-1000) were taken and powdered then concentration of drug solution for six times.
Asian Journal of Pharmacy and Pharmacology 2016; 2(1): 6-9 Limit of Detection and Limit of Quantitation
makes the results satisfactory. Notion at the back that effort Several approaches for determining the detection limit and was diminish the uncertain hindrances caused during the quantitation limit are possible, depending on whether the analysis. The data obtained for the estimation of metformin procedure is a non-instrumental or instrumental.
in bulk and marketed formulation proved the high level accuracy and precision after calibration.
Ruggedness of the proposed method is determined by analysis of Table 2. Results of recovery studies
aliquots from homogenous slot by two analyst using same % Amount of drug
% Recovery
% Amount of drug
% Recovery
operational and environmental conditions.
Results and discussion
The method was validated according to International Conference on Harmonization (ICH guideline, 2003). Q2B absolute guidelines for validation of analytical method for the determination of linearity, sensitivity, precision and accuracy.
Table 3. Results of precision studies (Intra-day and Inter-
Precision of the method was determined by adding known amounts of pure drug (80, 100 and 120%) in triplicate. In UV spectroscopic method, the zero and first order spectra were utilized for developing the equations for analysis. Metformin HCL showed maximum absorbance at 234 nm and 246 nm for zero and first order derivative spectroscopy respectively. The normal spectra were derivatized into first order derivative, using UV software of instrument, where Δλ = 2. The amplitudes of the corresponding troughs were measured at 246 nm (Table 1). The percentage recovery value obtained within standard limit of Table 4. Results of repeatability studies
99.43% to 101.2 % for both methods which confirmed that the methods were accurate and free from any interference of found (%)
taken (μg/ml) found (%)
excipients (Table 2). The low value of standard deviation obtained indicated precision of the method. Results of intraday and interday precision studies are reported in Table 3. The reproducibility, repeatability (Table 4) and ruggedness (Table 5) of proposed methods were found to be satisfactory which was Table 5. Results of ruggedness studies
evidenced by low values of standard deviation, LOD and LOQ Component Label claim
(Table 6) for both methods were found to be satisfactory.
Amount Found (%) (n=5)
Amount Found (%) (n=5)
Analyst I
Analyst II
Analyst I
Analyst II
Table 1. Linearity study of Metformin HCL
Table 6. Results of LOD and LOQ
(Mean ± SD)
(Mean ± SD)
The developed method which incorporates a change in the Present study showed the applicability of multivariate linear solvent used in the methods for the initial dilution of the regression move towards to the UV data obtained at different tablet samples would lead to accurate quantification of wavelengths for the calibration and tablet analysis. Statistically, metformin HCl. The developed method was successfully the use of vast number of data calculated for a sample analysis validated and should be used for the assay of metformin Asian Journal of Pharmacy and Pharmacology 2016; 2(1): 6-9 hydrochloride in metformin HCl tablets. Percent recovery and used in the management of non-insulin-dependent resulted concentration of active ingredient in formulations diabetes mellitus (NIDDM). Drugs, 49 (5): 721–49 showed that the amount of drug present is reliable with the label ICH guideline, 2003. Stability Testing of New Drug claim. Hence, this method is very useful with very mathematical Substances and Products Q1A (R2), International contents, is more consistent than the other spectrophotometric Conference on Harmonization, IFPMA, Geneva.
methods and strongly recommends the application in calibration Jani BR, Shah KV, Kapupara P. 2015. Development and models for a routine analysis.
validation of UV Spectroscopic method for simultaneous estimation of Dapagliflozin and Amruta B, Minal R, Ghante, S D Sawant. 2012. Simultaneous Metformin. International Journal of Research and UV spectrophotometric method for estimation of sitagliptin Development in Pharmacy and Life Sciences, phosphate and metformin hydrochloride in bulk and tablet dosage form Scholars Research Library, Der Pharma Ruggiero-Lopez D, Lecomte M, Moinet G, Patereau G, Chemica, 4 (3): 854-859.
Lagarde M, Wiernsperger N. 1999. Reaction of Arayne MS, Sultana N, Zuberi MH, Siddiqui FA. 2009.
metformin with dicarbonyl compounds. Possible Spectrophotometric quantitation of metformin in bulk drug implication in the inhibition of advanced glycation end and pharmaceuticalformulations using multivariate product formation. Biochemistry and Pharmacology 1, technique. Indian Journal of Pharmaceutical Science, 71(3): 58(11): 1765-73.
Setter SM, Iltz JL, Thams J, Campbell RK. 2003.
Chhetri HP, Thapa P, Schepdael AV. 2014. Simple HPLC-UV Metformin hydrochloride in the treatment of type 2 method for the quantification of metformin in human diabetes mellitus: a clinical review with a focus on dual plasma with one step protein precipitation. Saudi therapy. Clinical Therapeutics 25(12): 2991-3026.
Pharmaceutical Journal, 22 ), (5 483–487.
Susan ZY, Jack AY. 2014. Long-term Drug Treatment for Dunn CJ, Peters DH. 1995. The biguanide metformin Obesity: A Systematic and Clinical Review. JAMA 1, (dimethylbiguanide) is an oral antihyperglycaemic agent 311(1): 74–86.

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B816679j 435.454

www.rsc.org/csr Chemical Society Reviews Artemisinin and its derivatives: a novel class of anti-malarialand anti-cancer agentsw Devdutt Chaturvedi, Abhishek Goswami, Partha Pratim Saikia, Nabin C. Barua*and Paruchuri G. Rao Received 3rd February 2009First published as an Advance Article on the web 24th August 2009DOI: 10.1039/b816679j In this tutorial review, an effort towards presentation of a comprehensive account of the recentdevelopments on various kinds of artemisinin derivatives including artemisinin dimers, trimersand tetramers has been made and their efficacy towards malaria parasites and different cancercells lines was compared with that of artemisinins, and various other anti-malarial and anti-cancerdrugs. It is expected that this review will provide first-hand information on artemisinin chemistryto organic/medicinal chemists, and pharmacologists working on anticancer and anti-malarial drugdevelopment.

Microsoft word - send.doc

Second to Fourth Digit Length Ratio (2D:4D) and Adult Sex Hormone Levels: New Data and a Meta-Analytic Review Running title: 2D:4D and adult sex hormone levels [ Accepted for Publication in Psychoneuroendocrinology. Final Unedited Draft ] Johannes Hönekopp*, Luise Bartholdt Technische Universität Chemnitz, Institut für Psychologie, Wilhelm-Raabe Str. 43, D-09120