مقالات پذیرفته شده در نهمین کنگره بین المللی زیست پزشکی
Optimization of the reaction conditions for the removal of colored impurities in crude heparin using potassium permanganate
Optimization of the reaction conditions for the removal of colored impurities in crude heparin using potassium permanganate
Mahnaz Hadizadeh,1,*Shadi Abkhiz,2Davood Zare,3Maliheh Safavi,4
1. Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box 3353-5111, Tehran 33531-36846, Iran 2. Iran University of Medical Sciences, Tehran, Iran. 3. Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box 3353-5111, Tehran 33531-36846, Iran 4. Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), P.O. Box 3353-5111, Tehran 33531-36846, Iran
Introduction: Heparin is one of the most widely used drugs in the world. Heparin is known as a life-saving drug due to its anticoagulant properties, and its important roles in the treatment and prevention of many serious diseases and many other related uses, including kidney dialysis, heart attack, cardiac arrhythmia, acute coronary syndrome, pulmonary embolism, stroke, deep vein thrombosis, and blood clots. The main sources of pharmaceutical heparin production are animal tissues (mainly porcine and bovine). Crude heparin extracted from animal sources contains various impurities, including protein, nucleic acid, non-heparin glycosamines, and dyes. In the process of heparin purification, all these impurities are removed to obtain pharmaceutical heparin approved by FDA standards. The present study aimed to determine the optimal conditions for the decolorization of crude heparin using potassium permanganate
Methods: Crude heparin was extracted from bovine intestinal mucosa by enzymatic treatment (protease). After defatting by the Folch method, potassium permanganate (KMnO4) was used as a strong oxidizing agent to remove colored impurities present in crude heparin. Various factors, such as potassium permanganate concentration, solution pH, reaction time, and temperature were optimized to obtain pure heparin with high activity.
Results: The results showed that 1 g mol of KMnO4 is required to decolorize 100 million units of heparin salt. The time required for complete decolorization varies depending on the nature of the crude heparin salt being treated and is complete between 15 and 30 minutes after the addition of the permanganate solution. If the decolorization reaction time is prolonged, the activity of heparin decreases. The pH range in which the reaction is carried out is also very important. If the pH is significantly below 8.0, decolorization of heparin salt occurs with difficulty or not at all; on the other hand, if the pH is significantly above 8.5, decolorization is accompanied by a significant decrease in heparin activity due to the destruction of its structure. The temperature at which the reaction is carried out is also important. If the reaction is carried out at room temperature, the resulting manganese dioxide precipitates in the form of very fine particles and in a colloidal form, which are very difficult to separate by filtration. But if the reaction is carried out at high temperature, manganese dioxide precipitates in granular form and is easily removed from the product. In this study, it was found that the most suitable temperature for the decolorization reaction is 80-85 °C.
Conclusion: Overall, the results of this study showed that various factors, including sodium permanganate concentration, solution pH, reaction time, and temperature, play an important role in the efficiency of the decolorization process and obtaining heparin with high activity.