Degradation study of different brands Degradation study of different brands of Ceftriaxone injection available in Aden city

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 drug molecules. Five different brands of Ceftriaxone injection were collected from the market and exposed to various stress condition like alkaline, acidic, oxidative and thermal degradation .The degradation process was followed by using spectrophotometric analysis. All the antibiotic brands undergo forced degradation at conditions in different ranges and the maximum degradation observed in the basic medium.

hydrolysis, oxidation and thermal stress and to our knowledge this research is the first work performed with the purpose to compare the degree of forced degradation in five different brands of Ceftriaxone sodium injection (500mg) available in Aden market.
Five different brands of ceftriaxone 500mg injection were purchased from local Aden market (  Table 1 shows the manufacturing and expiry date of different brands),and the experimental work was done in College of Pharmacy, during July -September 2016 . Methods:

Preparation of Standard Stock Solution of Ceftriaxone Drug (5mg/ml)
The solution of ceftriaxone was prepared by taking 500mg of ceftriaxone injection dissolved in 5ml of sterile water and transferred into 100ml conical flask and the volume was made up to the mark with de-ionized water. Preparation of working Solution of Ceftriaxone Drug (1mg/ml) 50ml of Standard Stock Solution of Ceftriaxone (5mg/ml) was withdrawn from the above prepared solution and diluted to 250ml by de-ionized water .

Estimation of ceftriaxone concentrations
The ceftriaxone concentration before and after degradation was estimated by measuring Prussian Blue Formation by taking 1ml from drug solution under study in separated small flask , then 1ml of 0.02M FeCl3 and 0.5ml of potassium hexacyanide was added to drug solution and the volume was made up to 10ml with de-ionized water. Absorbance of the colored complex solution was recorded by spectrophotometer at the wavelength 700nm.

Studying the effect of acidic medium
10ml from the working solution of each brand was taken in small conical flasks then 10ml of 0.1M HCl was added in each conical flask and , after one hour ,the concentration of antibiotic was estimated. Studying the effect of basic medium 10ml from the working solution of each brand was taken in small conical flasks, then 10ml of 0.1M NaOH was added in each conical flask and , after one hour, the concentration of antibiotic was estimated. Studying the effect of oxidation 10ml the from working solution of each brand was taken in small conical flasks then 10ml of 3% H2O2 was added in each conical flask and , after one hour , the concentration of antibiotic was estimated. Studying the effect of heat 10ml from the working solution of each brand was taken in separated small conical flasks. then the solution was put in water bath 60 o C for a period of 1 hour. The study of the kinetic degradation was carried out by taking 10ml from the working solution of each brand was taken in Pyrex volumetric flask , then these solutions were placed in water bath for 2 hours at 60°C , and every 30 minutes 1ml of the antibiotic solution was withdrawn in separated test tube for further treatment to calculate the rate of degradation.

Results and discussion
The table (2) the results of ceftriaxone forced degradation indicates the effects of acidic medium ,basic medium and oxidative medium .The table shows the variation in absorbance before and after the effect of different degradation parameters after one hour exposure.

Acidic medium effect
The previous research (9) indicated that the fluoroquinolones degradation has happened due to the stress acidic by decarboxylation process. The maximum degradation for acidic pH effect was found in CTX4 and CTX3 which is equal to 47.96% and 43.67% respectively, but the minimum degradation was 9.68% , as shown by CTX1 (Fig.2

Basic medium effect
The alkaline degradation of the ceftriaxone, yielded to the opening of the main active bactericidal part of the antibiotic, the beta lactam ring (9) .The alkaline degradation of the ceftriaxone, yielded to the opening of the main active bactericidal part of the antibiotic, the beta lactam ring (9) .It was observed that the degree of the degradation due to the basic pH effect was found very high comparatively with acidic pH effect, the maximum percent revealed in CTX1 (81.25 %) and CTX2 (81.17%) , but the lowest value was 56.75% for CTX5 (Fig.3).

Oxidation effect
The oxidative degradation of drug substance involves an electron transfer mechanism to form reactive anions and cations. Amines, sulfides and phenols are susceptible to electron transfer oxidation to give N-oxides, hydroxylamine, sulfones and sulfoxide. The functional group with labile hydrogen , like benzylic carbon, allylic carbon, and tertiary carbon or α-positions with respect to heteroatom , is susceptible to oxidation to form hydroperoxides, hydroxide or ketone (8) . In case of the Ceftriaxone injection oxidation study by hydrogen peroxide, the degradation degree was found low ,it was 18.5% and 20.35% for CTX4 and CTX2 respectively , but the highest degradation registered in CTX1 brand was 52.43 % (Fig.4).

Fig.4: Effect of oxidation Heat effect :
The thermal degradation results (table 3) of different brands of ceftriaxone indicated that the absorbance decreased in all the brands and the maximum degradation was registered in CTX5 (67.47%) than the other antibiotic brands which resist the thermal degradation.

Conclusion:
All the antibiotic brands undergo forced degradation in different ranges and the maximum degradation has been observed in CTX4 , CTX1, CTX1 and CTX5 due to acid effect ,base effect ,oxidation effect and heat effect respectively.