UV Spectroscopy Analysis and Degradation Study of Rivaroxaban

 

Girishchandra R. Mandake^*, Indrajit S. Patil, Omkar A Patil, Dr. Manoj M. Nitalikar,

Dr. Shriniwas K Mohite

Rajarambapu College of Pharmacy, Kasegaon, Kasegaon, Walwa, Sangli, Maharashtra, India

 

ABSTRACT:

Rivaroxaban is a potent selective oral direct factor Xa inhibitor, which undergo hepatic first pass metabolism and high oral bioavailability. According to ICH guidelines, the major factors that contribute in degradation of a drug product comprise of temperature, time, photo degradation, pH variation (high and low), acid/base stress testing and/or with humidity. An attempt was made to examine and calculate the quantity of drug in the presence of degradation products by UV-Vi spectroscopy method. According to the WHO, the official assay limit of the content should not less than 97% and not more than 101.05% of labelled amount of Rivaroxaban. The results of experiment revealed that Rivaroxaban degrade much especially on exposure to UV light and heat but do not degrades in basic medium whereas slight degradation occurs in acidic medium.

 

 

 

INTRODUCTION:

Rivaroxaban is a potent selective oral direct factor Xa inhibitor, having low solubility and high permeability, which is used as an oral anticoagulant for the prevention of thromboembolism.(1) Rivaroxaban drug used in acute coronary syndrome (ACS), Prevention of cardiovascular death, myocardial infarction and stent thrombosis.(2) The effects last 8–12 hours, but factor Xa activity does not return to normal within 24 hours so once-daily dosing is possible.(3) Rivaroxaban is rapidly absorbed and achieves maximum concentration (Cmax) 2-4 h after tablet intake. Its oral absorption is close to 100% and its oral bioavailability is high (80-100%).(4) Peak effect of Rivaroxaban after oral administration is 2-4 hours and rapid onset of action with a half life of 5-9 hours.

 

On literature survey it was found that very few analytical methods are available for the estimation of Rivaroxaban. Rivaroxaban was subjected to acidic, basic, oxidative, photolytic, and thermal conditions for forced stress degradation studies. Considerable degradation was observed in all stress degradation tests. The chromatogram was recorded at 251 nm using a general ultraviolet (UV) detector. Rivaroxaban does not inhibit thrombin (activated Factor II). (5)

 

Rivaroxaban (RXN) is an orally active anticoagulant, direct factor Xa inhibitor approved for the prevention of venous thromboembolic events in patients who have undergone total hip or total knee replacement surgery. RXN blocks the amplification of the intrinsic and extrinsic pathway of coagulation cascade by binding directly to the catalytic pocket of factor Xa and thereby preventing the formation of thrombus. The half-life of RXN is 5–9 hrs in young subjects and 11–13 hrs in older subjects. Chemically, RXN is known as (S)-5-Chlor-N-{2-oxo-3-[4-(3-oxomorpholin-4-yl)phenyl]-1,3-oxazolidin-5-ylmethyl}thiophen-2-carbamid (Figure 1).(6)  

 

 

 

Figure 1: Chemical Structure of Rivaroxaban

 

 

UV spectrophotometry is widely used for the quantitative analysis of the active substances in dissolution test samples.(7) Rivaroxaban, an amide group containing oral anticoagulant was subjected to stress conditions commonly required for the registration of pharmaceuticals: base and acid-catalyzed hydrolysis (0.1 M, 60 °C, 3–6 h), oxidation (10% H₂O₂, 24 h), photodegradation (300–800 nm, 24 h) and thermal decomposition (50 °C, 6 h).(8) Spectrophotometry is the most preferred technique for degradation studies over other methods because of less equipment cost and economical maintenance advantage. Most of the reported methods either do not include stress degradation studies or are not completely validated and they are time consuming and expensive. Spectrophotometry technique is based on measuring the absorption of a monochromatic light in the near ultraviolet region (200-380nm) by colorless complex. UV spectrophotometry can also be used for stress degradation.(9) According to International Conference of Harmonization (ICH) guideline active pharmaceutical ingredient is focused to various forced degradation conditions which are acidic, basic and light conditions. Degradation studies are necessary in the development of new drug product. Many factors are responsible for degradation of drug or product like temperature humidity, and light. In this research work the effect of various factor like acid, base, UV light and heat on Rivaroxaban was studied. UV spectrophotometry is widely used for the quantitative analysis of the active substances in dissolution test samples. In this study, a UV spectrophotometric method was developed for the determination of the release of rivaroxaban (Figure 1), a new anticoagulant, from a commercially available tablet dosage form (Xarelto-10mg). According to the literature there are two different HPLC methods available for the determination of rivaroxaban in tablets. In the present study, a UV spectro‑ photometric method was developed to determine the rivaroxaban content in pharmaceutical formulations and the amount released from rivaroxaban tablets in dissolution studies. Validation for the proposed methods was done as per International Conference on Harmonization (ICH) guidelines.(10)

Parameters Involved in Forced Degradation

Distinctive forced degradation studies on drug substance involves acid/base stress testing, photo degradation, temperature, time, pH variation (low and high).

 

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, esters (lactones) aryl amines, carbomets acid 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 expose to continuous sunlight. The degree of disintegration is depends up on degree of exsposer.

 

Thermal (Heat) and / or humidity stress testing

Many drugs are thermal (heat) and humidity sensitive. Thermal (Heat) and humidity is degrade substance up to its main components. This test is performed by exposing the drug or formulation to thermal / humidity condition.

 

EXPERIMENTAL:

Material and Reagents:

Pure Rivaroxaban was procured as a purchase from Cipla, Mumbai. products and all other chemicals used were of AR grade. UV-visible spectrophotometer (JASCO V-630) with quartiz cuvett was used for the studies.

 

0.1 N hydrochloric acid was prepared by taking 8.3 ml hydrochloric acid (HCl) in 100 ml volumetric flask having purity 37% and the final volume upto the mark of the flask was made with distilled water. Same procedure was followed for the preparation of 0.1N sodium hydroxide solution. 200 ppm Rivaroxaban solution was prepared by accurately weighing 0.020gm pure Rivaroxaban and was transferred it into 100ml volumetric flask. Small amount of water was added in volumetric flask to dissolve Rivaroxaban and final volume was adjusted with distilled water. Absorbance of solution of Rivaroxaban was measured by UV spectrophotometer at maximum wavelength 251 nm.

 

Degradation studies:

For acid:

To study degradation in acid medium or to determine effect of acid on Rivaroxaban, 5ml of 200ppm Rivaroxaban solution was mixed with 5ml of 0.1 N hydrochloric acid (HCl) solution and was kept aside for 30 minutes. After 30 minutes absorbance of solution was measured by UV spectrophotometer at maximum wavelength 251nm.

 

For base:

To study degradation in basic medium or to determine effect of base on Rivaroxaban, 5ml of 200ppm Rivaroxaban solution was mixed 5ml of 0.1 N sodium hydroxide solution (NaOH) was kept aside for 30 minutes. After 30 minutes absorbance of solution was measured by UV spectrophotometer at maximum wavelength 251nm.

 

For UV light:

To study degradation in UV light medium or to determine effect of UV light on Rivaroxaban, 5ml of 200ppm Rivaroxaban solution in test tube and 5ml distilled water was added and kept aside for 30 minutes in UV light of 320nm. After 30 minutes absorbance of solution was measured by UV spectrophotometer at 251 nm.

 

For thermal (Heat):

To study degradation in thermal medium or to determine effect of heat on Rivaroxaban, 5 ml of 200 ppm Rivaroxaban solution was taken and 5ml of distilled water was added and was kept aside for 30 minutes in water bath at 50⁰ c. After 30 minutes absorbance of solution was measured by UV spectrophotometer at maximum wavelength 251 nm.

 

RESULTS AND DISCUSSION: 

Rivaroxaban was exposed to various physical conditions and degradation effect occurred was studied by UV. Results obtained are mentioned. Absorbance after exposure to different conditions was measured and results are shown in table 1 and shown graphically in fig. 1. Percentage degradation was calculated and is shown in table 2 and graphically in fig. 2.

 

When Rivaroxaban was exposed to 0.1N HCl, Rivaroxaban did not show any significant change in terms of availability (68.17%). But, when Rivaroxaban was exposed to 0.1 N NaOH, Rivaroxaban showed highly significant change in term of availability (107.31%). When Rivaroxaban exposed to heat for 30 minutes, highly decreased availability observed (79.04%) and when exposed to U.V light, Rivaroxaban also showed decreased availability (95.27%).

 

It can be concluded that Rivaroxaban does not get degraded on exposure to acidic medium 0.1N HCl. (68.17 %) but have major degradation effect on exposure to basic medium 0.1N NaOH (107.31%). When Rivaroxaban was exposed to UV light and heat for 30 minutes high degradation was observed 79.04% and 95.27% respectively.

Table 1: Absorbance of  Rivaroxaban

                  

 

 Figure 2: Absorbance of Rivaroxaban

 

 

 

Table 2: Degradation Pattern in Percentage of Rivaroxaban

Degradation Parameters	Readings after 30 Minutes
	1	2	3	Average
Before	100	100.04	100.01	100.01
After acid	68.16	68.20	68.17	68.17
After base	107.31	107.33	107.31	107.31
After heat	79.04	79.03	79.06	79.04
After U.V	95.28	95.27	95.28	95.27

  

 

 

Figure3: Degradation Pattern of Rivaroxaban

 

CONCLUSION:

According to specification given in WHO monographic for Rivaroxaban, the official assay limit of the content should not less than 99% and not more than 102% of the estimated potency.  From our result we concluded that Rivaroxaban degrades to a very larger extend when exposed to basic medium whereas it also degrades in the presence of acidic medium, heat and UV light.

 

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Accepted on 28.04.2018            © A&V Publications All right reserved