Preparation Of Pyroline Praeparatum And Establishment Of Its Quality Standard

Pyroline is a new veterinary drugs with high efficiency, broad spectrum, low toxicity and no residual for treatment of bovine mastitis, bowel oedema disease, bovine endometritis, pullorum disease, swine yellow dysentery, swine white dysentery and swine comprehensiveness diarrhea. The drug is white crystal like lamellar or needle, easily dissolved in water,ethylether and other organic solvent. Because the measure of recrystallization used in the industrialized manufacture of pyroline is harmful to those who synthesis the drug and the praeparatum of pyroline used at present is easy to be oxidated to decrease its potency. So we improve its technology flowsheet in order to fit industrialized manufacture and prepare pyroline's new praeparatum, and study on its quality control & stability ,all of these above will supply the scientific bases to its formal use in veterinary clinic.1. The improvement of the technology flowsheet of pyroline' industrialized manufacture: Firstly put the o-toluidine and manganese dioxide together at room temperature catalysed by sulphuric acid and then get the admixture of orth-methyl benzoquinone, Secondly aqueous solution of orth-methyl benzoquinone is got through wet distillation, Thirdly the admixture of pyroline is got after reducted by sodium dithionite, then pharmaceutical product of pyroline is got through extracted and concentrated by ethylether and recrystalled by 1% sodium bisulfite.The results showed that this synthesis procedure is easily operation, safe and low cost compared with other methods.2. The preparation of praeparatum for pyroline: Pyroline for injection No.1 is got through drying, Weighing, subpackage, ultraviolet lamp sterilizing and sealing of the pharmaceutical product of pyroline; Put sodium bisulfite and edathamil disodium to the pharmaceutical product of pyroline get pyroline for injection No.2; Put sodium bisulfite and edathamil disodium to the aqueous solution of pharmaceutical product of pyroline get pyroline injection.3. Establishment of a method for determining pyroline content:RP-HPLC was carried out with the internal standard 1, 4-dihydroxybenzene. Symmetry (?)C₁₈ column( 4.6mm×150mm, 5μm) ,the detection wavelength atλ_mas= 302nm,The mobile phase was composed of CH₃OH:H₂O(20:80), The flow rate was 0.8ml/min. The results showed the liner relationship was presented over the range of 60μg/ml～140μg/ml with equation Y= 0.0086X+0.00416( r = 0.9999). The proposed method is simple,accurate and precise which can be applied in the determination of the content for pyroline for injection No. 1, pyroline for injection No.2 and pyroline injection. 4. The quality standard investigation of praeparatum for pyroline: The quality standard investigation for injection includes clarity,asepsis,osmotic pressure, haemolyticus, acidity,localized stimulus, pyrogen, anaphylactic response and assay. The results showed that pyroline for injection No.1 and pyroline for injection No.2 were white crystal, It is clear,no particle, innoxious, asepsis, moderate osmotic pressure, no haemolysis,PH6.1, no stimulus, nonpyrogenic,no allergy and stable. The quality of pyroline injection was also fit the common requirement of injection.5. The study on the stability of praeparatum for pyroline and its expiration date calculation: The stability test includes influential factor test, accelerated test and long-term test.(1)The results showed that the content of pharmaceutical product of pyroline were 98.15 % after 5 days and 98.23% after 10 days respectively under the high temperature(60℃) with no significant difference on aspects of color, pH and clarity. (2) The content of pharmaceutical product of pyroline were 94.81 % after 5 days and 89.03 % after 10 days respectively under the high humidity(relative humidity 90%±5%), room temperature away from light, with color changed from achromatism to pink, the content decreased obviously, moisture regain weight gain.(3) The content of pharmaceutical product of pyroline were 98.15 % after 5 days and 97.83% after 10 days respectively under the strong light-strucking (the illuminance is 45001x±5001x), room temperature ,dry condition ,with no significant difference on aspects of color, pH and clarity.The results showed that the pharmaceutical product of pyroline is stable.(4)The content of aqueous solution of pharmaceutical product of pyroline were 79.15% after 5 days and 60.83 % after 10 days respectively under the high humidity(relative humidity 90%±5%), room temperature away from light, with color changed from achromatism to pink and then to brown or black, the longer time it was placed the more muddy it was. The results showed that the pharmaceutical product of pyroline is instability at the aqueous solution. (5) Put the pyroline for injection No.1 , pyroline for injection No.2 and pyroline injection under the accelerated condition(temperature 40±2℃, relative humidity 75%),six months later the content ,aspects of color, pH and clarity have no significant change. the results showed that the pyroline for injection No.1 , pyroline for injection No.2 and pyroline injection is stable under the accelerated condition. (6) Put the three batches of pyroline for injection No.1 , pyroline for injection No.2 and pyroline injection for 1.5 years under storage condition( temperature 25±2℃, relative humidity 60±5%),after statistical analysis and calculation we get the conclusion that the term of validity of pyroline for injection No.l, pyroline for injection No.2 and pyroline injection is 29.5,36,24.2 monthes respectively, the results showed that the pyroline for injection No. 1 , pyroline for injection No.2 and pyroline injection is stable under the storage condition.