INTRODUCTION:

Analytical chemistry may be derived as the science and art of determining the composition of material in terms of the elements of compounds contained. By means of analytical techniques both qualitative analysis (the presence or absence of one or more elements) and quantitative analysis (how much amount is present) can be done.

Pharmaceutical analysis may be defined as the application of analytical procedures used to determine the purity, safety and quality of drugs and chemicals. It includes both qualitative and quantitative analysis of drugs and pharmaceutical substances start from bulk drugs to the finished dosage form¹.

HYDROTROPHY:

Hydrotropy could be a distinctive and unprecedented solubilization technique in which certain chemical elements termed as hydrotropes can be used to effect a several fold increase within the solubility of meagerly soluble solutes under normal conditions². This increase in solubility in water is probably due to the formation of organized assemblies of hydrotrope molecules at critical concentration. Hydrotropes in general are water soluble and surface-active compounds, which might considerably enhance the solubility of organic solutes like acids, esters, alcohols, aldehydes, ketones, hydrocarbons and fats. Hydrotropes are widely used in drug solubilization, detergent formulation, health care and household applications as well as for being an extraction agent for fragrances.

Mixed hydrotropy:

Mixed hydrotropic solubilization technique³ is the phenomenon to increase the solubility of poorly water-soluble drugs in the blends of hydrotropic agents, which can provide miraculous synergistic enhancement effect on solubility of poorly water soluble drugs, utilization of it within the formulation of dosage forms of water insoluble drugs and to reduce concentration of individual hydrotropic agent to minimize the side effects (in place of using a large concentration of one hydrotrope a blend of, say, 5 hydrotropes are often used in 1/5th concentrations reducing their individual toxicities.

Advantages of hydrotropic solubilization:

It may reduce the large total concentration of hydrotropic agents necessary to produce modest increase in solubility by employing combination of agents in lower concentration. It precludes the use of organic solvents and thus avoids the problem of residual toxicity, error due to volatility, pollution, cost etc. The use of hydrotropy to give fast release of poorly water-soluble drugs from the suppositories. Hydrotropic solutions can also be tried to develop the dissolution fluids to carry out the dissolution studies of dosage forms of poorly water-soluble drugs.

Gliclazide comes under the category of sulfonyl ureas used in the treatment of diabetes. Gliclazide can be estimated by various methods like UV, HPLC by utilizing various organic solvents to solubilize the drug. An attempt has been made to solubilize the drug ecofriendly by application of hydrotropic solubilization technique, an alternative method for the estimation of drug in dosage form.

MATERIALS AND METHODS:

CHEMICALS AND REAGENTS:

Gliclazide pure drug sample was obtained from Yarrow chem. products Mumbai. Commercial tablet Diamicron-80 mg was purchased from local market. Urea, Tri Sodium citrate, sodium benzoate, sodium acetate used was of S.D Fine Chemical Limited, Mumbai.

Instrumentation:

the proposed work was carried out on an analytical UV- visible spectrophotometry (Analytical 2060 plus Model), which possesses a double beam double detector configuration with a 1 cm quartz matched cell. All weighing was done on electronic precision balance (Contech-Citizen Sales Private Limited.)

SELECTION OF HYDROTROPIC SOLUBILISING ADDITIVE:

Various solubilising additives like sodium acetate, niacinamide, sodium benzoate, urea, trisodium citrate were tried. Sodium acetate, niacinamide on addition to drug causes insolubility. Sodium benzoate leads to sedimentation of drug. Blends of urea (6M) and trisodium citrate (2M) renders complete solubilization of drug.

PREPARATION OF CALIBRATION CURVE OF GLICLAZIDE:

100 mg of drug was weighed accurately and transferred into 100 ml volumetric flask. To this, 80 ml of 6M urea and 20 ml of 2M trisodium citrate solution was added and shaken for about 15 minutes to solubilized the drug and final volume was adjusted with distilled water to 100ml.(1mg/ml). Then pipette out 1 ml from the stock solution and diluted to 10ml with distilled water to obtain a concentration of 100 µg/ml. Various dilutions containing concentrations of 4-12 µg/ml solutions are prepared. The absorption maxima (λ max) of Gliclazide were also found.

ASSAY OF GLICLAZIDE TABLETS:

Weigh 20 tablets, powdered and weigh accurately a quantity equivalent to 100 mgof drug from the tablet powder and transferred into 100 ml volumetric flask. To this, 80 ml of 6M urea and 20 ml of 2M trisodium citrate solution was added and shaken for about 15 minutes to solubilized the drug and final volume was adjusted with distilled water to 100ml.(1mg/ml). Then pipette out 0.4 ml of solution and make up to 10ml leads to 8μg/ml concentration solution. The absorbance of the resulting solution was measured at 225 nm against solvent blank and the drug content was calculated.

VALIDATION OF THE PROPOSED METHOD:

The proposed method was validated⁴ for the following parameters.

Precision:

Precision was determined by studying the repeatability and intermediate precision. The standard deviation, coefficient of variance and standard error were calculated for the drug.

Inter- day and Intra- day precision:

The intra-day concentration of the drug was calculated on the same day at an interval of two hours, whereas the inter day concentration of drug was calculated on three different days within the laboratory conditions.

Linearity:

The absorbance of appropriate dilutions of standard stock solutions was measured as per the developed method to confirm the linearity.

Limit of detection (LOD) and Limit of Quantitation (LOQ):

The LOD and LOQ of Gliclazide by the proposed method were determined using calibration standards. LOD and LOQ were calculated as 3.3σ /S and 10σ/S, respectively, where S is the slope of the calibration curve and σ is the standard deviation of response

Accuracy:

Accuracy is the percentage of analyte recovered by assay from known added amount. Analysis was performed at 80%, 100% and 120% levels.

Recovery studies:

In order to check the accuracy and reproducibility of the proposed method, recovery studies were conducted. Recovery studies are done spiking method in this method the test sample having the concentration of 8μg/ml. To this the standard drug is spiked by adding into the test solution. A concentration of 6, 8 and 10μg/ml are added to the sample solutions and the absorbances of the three spiked concentrations were taken. From this absorbance we can determine the amount of drug that can be recovered by the proposed method.

RESULTS:

Figure 1. Lambda max (λ max) of Gliclazide

Table No: 1 Calibration values of Gliclazide

S. No	Concentration (µg/ml)	Absorbance
1	4	0.206
2	6	0.302
3	8	0.380
4	10	0.486
5	12	0.599

Figure 2. Calibration curve of Gliclazide

Table no: 2 Method validation parameters

S. No

Parameters

Results

Lambda max

Beers law limit

Regression eq;( y=mx+c)

Slope

Intercept

Correlation coefficient

Relative standard deviation

LOD value

LOQ value

225 nm

4-12 µg/ml

0.048(x)+0.006

0.048

0.006

0.995

0.644

0.066

0.198

Table no: 3 Precision Studies

Sample (µg/ml)	Intraday precision		Inter day precision
Sample (µg/ml)	Absorbance	RSD	Absorbance	RSD
6	0.356 0.380 0.325	0.0779	0.380 0.335 0.350	0.0645
8	0.467 0.436 0.442	0.0366	0.434 0.413 0.427	0.0251
10	0.588 0.574 0.568	0.0177	0.532 0.491 0.512	0.0400

Table no: 4 Recovery Studies

Test µg/ml

Amount of standard drug added µg/ml

% Recovery of Gliclazide

Standard deviation

RSD

8µg/ml

95.33%

96.22%

95.55%

0.4635

0.0048

Table no: 5 Analysis of Tablet Formulation of Gliclazide

Drug	Label claim(mg)	Amount found (mg)	% Purity
Gliclazide	80 mg	77.08 mg	96.35

DISCUSSION:

The solubility studies showed that aqueous solubility of Gliclazide was increased by adding tri sodium citrate (2M) and urea (6M) as hydrotropic solubilising agent. The Beer- Lambert’s concentration range for Gliclazide was between 4-12 µg/ml having absorption maxima at the wavelength of 225 nm.

The recovery studies showed proposed method is accurate and reproducible. The results of recovery study revealed that any small change in the drug concentration in the solution could be accurately determined by the proposed method. Accuracy, reproducibility and precision of the proposed methods were further confirmed by percent recovery values, which were close to 100 with low values of standard deviation as shown in Table no: 4

Repeatability results indicated the precision under the same operating conditions over a short interval time and inter assay precision. Intermediate precision study expresses within laboratory variation in different days. In both intra and inter-day precision study for the method RSD were not more than 1.0% indicates good intermediate precision. The low values of LOD and LOQ, 0.066 and 0.198 for Gliclazide indicate good sensitivity of proposed method. (Table no: 2).

CONCLUSION:

This current investigation is intended to use hydrotropy technique in UV spectrophotometric method to determine the assay of Gliclazide. The proposed method was found to be novel, ecofriendly, simple, precise, specific and highly accurate. This can be employed for routine analysis of Gliclazide in oral and bulk formulations.

ACKNOWLEDGEMENT:

The authors acknowledge the management of the Jyothishmathi College of Pharmacy, Turkapally to provide facilities to perform this research work and promote the method evaluation of the same.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 07.08.2018 Accepted on 22.10.2018

Asian J. Pharm. Ana. 2019; 9(2):45-48.

DOI: 10.5958/2231-5675.2019.00010.3