Degradation Study of Furosemide by UV- Visible Spectrophotometry Method in bulk form

 

Rohankumar R. Chavan*, Somnath D. Bhinge, Mangesh A. Bhutkar, Dheeraj S. Randive

Dept of Pharmaceutical Chemistry, Rajarambapu College of Pharmacy, Kasegaon,

Dist – Sangli, Maharashtra, India – 416404

*Corresponding Author E-mail: rohankumar3102@gmail.com

 

ABSTRACT:

Furosemide, a widely used “high-ceiling” loop diuretic drug, is indicated for congestive heart failure, chronic renal failure, and hepatic cirrhosis. Degradation is a main problem in numerous unstable products. As per the ICH guidelines factors which causes degradation of a drug product comprise of temperature, time, photo degradation, pH variation (high and low), acid/base stress testing and/ or with humidity. In our research work UV-V spectroscopy method was designed to examine and calculate the quantity of drug in the presence of degradation products. WHO, the official assay limit of the content should not less than 98% and not more than 102% of labelled amount of the estimated potency. From the results of our study it can concluded that Furosemide degrades most when exposed to the acidic medium and heat, whereas slight degradation occurs in basic medium and UV light.

 

KEYWORDS: Furosemide, Degradation, UV-Spectrophotometry.

 

 


INTRODUCTION:

Furosemide (FUR), is chemically 5-(aminosulfonyl)-4-chloro-2-[(2 furanylmethyl) amino] benzoic acid shown in Fig. 1.The molecular formula is C12H11ClN2O5 S and molecular weight is 330.745g/mol. It is soluble in acetone, methanol, DMSO, and alkali hydroxides and practically insoluble in water. [1] It is a loop diuretic that has been used in the treatment of congestive heart failure and edema. FUR acts on thick ascending limb of the loop of Henle leading to a loss of sodium, potassium, and chloride that are dispatched in the urine. [2] It results in a decrease in sodium and chloride reabsorption, while increasing the excretion of potassium in the distal renal tubule.

 

Fig. 1. Structure of Furosemide

 

The purpose of stability testing is to provide evidence on how the quality of a drug substance or drug product varies with time under the influence of a variety of environmental factors. [3] Initial improvement process, forced degradation activities must be executed to ensure that the process is selective to save lot of time, effort, money and to find out the responsible conditions for drug degradation. [4]

The main reason of degradation are the chemical instability of the drug substance under the conditions of heat, humidity, solvent, pH, and light encountered during manufacture, isolation, purification, drying, storage, transportation, and/or formulation is main cause of its degradation. [5]

 

The method of analysis is based on the measuring in the near (UV) ultraviolet path of spectrum (200-400 nm). UV spectrophotometry can be used for stress-degradation studies of Furosemide. The active pharmaceutical ingredient is subjected to a number of forced degradation conditions to include acidic, basic and photo conditions as per ICH guidelines (Q1 A and Q1B) [5-6]. Forced degradation should be one of the activities performed early in the development process to ensure that the method is discriminating before a lot of time, effort, and money have been expended. It is important to determine the conditions responsible to degrade the drug. Literature survey revealed that several photodegradation, [7] High-performance liquid chromatography (HPLC) [8] methods for stability study of furosemide but no report is available for degradation studies of Furosemide by UV.

 

Thus, in the present study Ultraviolet-visible (UV-Vis) spectroscopy was used for the analysis of forced degradation of Furosemide, as it is a simple, rapid and cost effective method as compared to HPLC.

 

Factors affecting forced degradation:

The factors which affect the API for forced degradation are temperature, humidity, acid/base stress testing, pH variations and photo degradation.

 

Thermal and/or Humidity Stress Testing:

Thermal and/or humidity stress testing is performed by keeping the drug substance to thermal/humidity conditions for extended period of time which causes the substance to degrade forcefully to its primary constituents components.

 

Acid/Base Stress Testing:

Acid/base stress testing is used for the evaluation of forced degradation of a drug substance. This test involves degradation of a drug substance by exposure to basic or acidic medium over time to its primary degradation products. In carbonyl functional group like alcohol, imines, imides, amides (lactums), esters (lactones) aryl amines and base degradation take place by hydrolysis.

 

Degradation by UV Light:

Many formulation or products made of synthetic or natural polymer are UV unstable. They degrade or disintegrate on its exposure to continuous sunlight. The degree of degradation or disintegration is dependent upon degree of exposure.

 

MATERIALS AND METHODS:

Chemicals and reagents:

Furosemide was kindly supplied as a gift sample by Yarrow Chemical Productions Mumbai (INDIA) 421201.

 

Sodium hydroxide (NaOH), Hydrochloric acid (HCL) and Double distilled water was procured from Research lab Islampur (INDIA) 415409. All chemicals and reagents were of analytical grade (AR).

 

Instrumentation:

An UV-Visible double beam spectrophotometer JASCO V 630 made in Japan with 1 cm matched quartz cells were used. All weighing operations were done on an electronic balance (Model Shimadzu AUW-220D), Ultrasonicator model 5.0L150H was used.

 

Glasswares:

All the glass material used in this research work namely test tube, beaker, measuring cylinder, funnel, pipette, volumetric flask and stirrer was made up of Pyrex glass. All the glass ware was first washed by chromic acid then rinsed with water and finally with distilled water.

 

Preparation of solutions:

Preparation of 0.1 N Hydrochloric Acid:

0.1 N hydrochloric acid was prepared by taking 8.3 ml hydrochloric acid in 100 ml volumetric flask having purity 37% and final volume was adjusted with distilled water.

 

Preparation of 0.1 N Sodium Hydroxide:

Accurately weighed 4 gm of sodium hydroxide (NaOH) was taken and transferred to a 100ml volumetric flask. Small amount of water was added to it to dissolve NaOH and final volume was made up with distilled water.

 

Preparation of Furosemide Stock Solution (Bulk drug):

Accurately weighted 10mg of Furosemide was transferred into 100 ml volumetric flask and double distilled water was added to make up the volume. The strength of solution was 10μg/mL-1. The absorbance of prepared solution of Furosemide was measured using a UV spectrophotometer at a maximum wavelength 276 nm. The tests were performed in triplicate.

 

Methods of Degradation Studies:

For UV light:

5 ml of 100 ppm solution of Furosemide was taken in test tubes and 5 ml of double distilled water was added to it and the tubes were kept in UV light for 30 minutes. After 30 minutes of UV exposure (243nm) the absorbance of the solutions was analyzed at wavelength 276 nm. The test was performed for three times.


 

Fig.2. Absorbance of Furosemide (API).

 

 

Fig.3. Percentage Degradation Pattern of Furosemide (API).

 


For Acid:

5 ml of 100 ppm solution of Furosemide was taken in a test tube, 5 ml of 1 N hydrochloric acid was added to it and kept aside for 30 minutes. After 30 minutes the absorbance of the solutions was analyzed separately at wavelength 276 nm. The test was performed for three times.

 

For Base:

5 ml of 100 ppm solution of Furosemide was taken in a test tube, 5 ml of 1N sodium hydroxide was added to it and kept aside for 30 minutes. After 30 minutes the absorbance of the solution was analyzed at wavelength 276 nm. The test was performed for three times.

 

For Heat:

To study degradation in thermal medium or to determine the effect of heat on Furosemide, 5ml of 100ppm Furosemide solution was taken and 5ml of distilled water was added to it and the mixture was left for 30 minutes in water bath at 50°C. After 30 minutes absorbance of the solution was measured by UV spectrophotometer at maximum wavelength 276 nm.

 

RESULTS AND DISCUSSION:

A degradation study is an essential parameter in the new drug development studies. The purpose of these degradation studies is to ensure the qualities of individual drugs as well as in combination of drug product, during its storage. The qualities of each drug component changes with the time under the exposure of environmental condition such as humidity, temperature and light. The purpose of degradation studies is to investigate those changes, to get a shelf life for the drug product and to find out storage conditions, which will be applicable to all future batches of the tested drug product manufactured and packaged under similar purpose.

 

Table 1. Absorbance of Furosemide (API)   

Degradation

Parameters

FUROSEMIDE

1

2

3

Mean

Before

0.8331

0.8345

0.8331

0.8335

After Acid

1.8413

1.8365

1.8412

1.8396

After Base

0.9373

0.9342

0.9362

0.9359

After Heat

1.6377

1.6245

1.6301

1.6307

After UV

0.4006

0.4106

0.3990

0.4034

 

 

Table 2. Percentage Degradation Pattern of Furosemide (API)

Degradation

Parameters

FUROSEMIDE

1

2

3

Mean

Before

100%

100.16%

100%

100.05%

After Acid

221.01%

221.0%

220.44%

220.82%

After Base

112.50%

112.13%

112.37%

112.33%

After Heat

196.57%

194.99%

195.66%

195.74%

After UV

48.05%

49.28%

47.89%

48.41%

 

We studied degradation parameters on active Furosemide. The absorbance for degradation parameters are given in Table 1. and shown in Fig.2. It is observed that Furosemide when subjected to acidic, basic medium and heat it showed increased availability i.e. 220.82%, 195.74%, 112.33% respectively and in the presence of UV light it showed 48.41% availability as shown in Table 2. and Fig.3.

 

Thus, from our results it can concluded that Furosemide when introduced in the basic medium i.e. 0.1N NaOH, it undergoes degradation up to 112.33%, whereas on its exposure to acidic medium it undergoes degradation to an extent of 220.82%. In case of UV light degradation studies, Furosemide when exposed to U.V. light showed 48.41% degradation, whereas on its exposure to heat for 30 min it exhibited 195.74% degradation.

 

CONCLUSION:

According to WHO monographic specifications the official assay limit of content should NLT 98% and NMT 102% of the estimated potency. The conclusion of our results is that the drug Furosemide degrades more when exposed to the acidic medium and heat, whereas slight degradation occurs when exposed to basic medium and U.V. light.

 

ACKNOWLEDGMENT:

Authors are thankful to Dr. C. S. Magdum, Principal, Rajarambapu College of Pharmacy, Kasegaon (Maharashtra) for providing necessary facilities to carry out the research work.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest

 

REFERENCES:

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2.      Giebisch G, The use of a diuretic agent as a probe to investigate site and mechanism of ion transport processes. Arzneimittel Forschung. 1985; 35(1); 336–342.

3.      ICH Topic Q 1A (R2), Stability Testing of new Drug Substances and Products. European Medicines Agency. 2003; CPMP/ICH/2736/99.

4.      Naveed S., Waheed N., Nazeer S, Degradation Study of Metronidazole in Active and Different Formulation by UV Spectroscopy. Journal of Bioequivalence & Bioavailability. 2014; 6; 124-127.

5.      ICH, Stability Testing: Photostability Testing of New Drug Substances and Products Q1b adopted in 1996, in Proceedings of the International Conference on Harmonization, Geneva, Switzerland, 2005.

6.      Hotha KK. Reddy SPK. Raju VK. Ravindranath LK. Forced Degradation Studies; Practical Approach- Overview of Regulatory Guidance and Literature for The Drug Products and Drug Substances. International Research Journal of Pharmacy. 2012; 4(5), 78-85.

7.      Bundgaard H., Norgaard T., Nielsen NM. Photodegradation and hydrolysis of Furosemide and Furosemide esters in aqueous solutions. International Journal of Pharmaceutics. 1988; 42 (1–3); 217-224.

8.      Tandel JN, Development and Validation of RP-HPLC Method for the Simultaneous Determination of Amiloride Hydrochloride and Furosemide in Pure and Pharmaceutical Dosage Form. Eurasian Journal of Analytical Chemistry. 2017; 12(4); 385-394.

 

 

 

Received on 22.02.2018            Modified on 07.05.2018

Accepted on 10.07.2018            © A&V Publications All right reserved

Asian J. Res. Pharm. Sci. 2019; 9(3):163-168.

DOI: 10.5958/2231-5659.2019.00025.0