Effect of Glycerol, as Cryoprotectant in the Encapsulation and Freeze Drying of Microspheres Containing Probiotic Cells

Oana Lelia Pop, Zorița Diaconeasa, Brandau Thorsten, Oana Ciuzan, Doru Pamfil, Dan Cristian Vodnar, Carmen Socaciu


It is reported that probiotics provide several health benefits as they help in maintaining a good balance and composition of intestinal flora, and increase the resistance against invasion of pathogens. Ensuring adequate dosages of probiotics at the time of consumption is a challenge, because several factors during processing and storage affect the viability of probiotic organisms. Major emphasis has been given to protect the microorganisms with the help of encapsulation technique, by addition of different protectants. In this study, probiotic cells (Bifidobacterium lactis 300B) were entrapped in alginate/pullulan microspheres. In the encapsulation formula glycerol was used as cryoprotectant in the freeze drying process for long time storage. It was observed that the survival of Bifidobacterium lactis 300B when encapsulated without cryoprotectant was higher than the formula with glycerol in the fresh obtained microspheres. The addition of glycerol was in order to reduce the deep freezing and freeze drying damages. In the chosen formulations, glycerol did not proved protection for the entrapped probiotic cells in the freeze drying process, for which the use of glycerol as cryoprotectant for alginate/pullulan Bifidobacterium lactis 300B entrapment is not recommended.


probiotic, encapsulation, freeze- drying, cryoprotectants, glycerol

Full Text:



Amine KM, Champagne CP, Salmieri Sp, Britten M, St-Gelais D, Fustier P Lacroix M (2014). Effect of palmitoylated alginate microencapsulation on viability of Bifidobacterium longum during freeze-drying. LWT- Food Sci Technol 56(1):111-117.

Anal AK Singh H (2007). Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery. Trends Food Sci Tech 18(5):240-251.

Aste T Weaire D, (2008). The Pursuit of Perfect Packing, Second Edition (2nd ed). Taylor and Francis group, Northwestern.

Augustin MA Sanguansri L, (2003). Encapsulation of food ingredients.

Benita S, (2006). Microencapsulation –Methods and Industrial Applications (2nd ed. ed). Taylor & Francis.

Burgain J, Gaiani C, Linder M Scher J (2011). Encapsulation of probiotic living cells: From laboratory scale to industrial applications. J Food Eng 104(4):467-483.

Capela P, Hay TKC Shah NP (2006). Effect of cryoprotectants, prebiotics and microencapsulation on survival of probiotic organisms in yoghurt and freeze-dried yoghurt. Food Res Int 39(203–211.

Chan ES Zhang Z (2005). Bioencapsulation by compression coating of probiotic bacteria for their protection in an acidic medium. Process Biochemistry 40(10):3346-3351.

Chavarri M, Maranon I, Ares R, Ibanez FC, Marzo F Villaran MdC (2010). Microencapsulation of a probiotic and prebiotic in alginate-chitosan capsules improves survival in simulated gastro-intestinal conditions. Int J Food Microbiol 142(1-2):185-189.

Cook MT, Tzortzis G, Charalampopoulos D Khutoryanskiy VV (2012). Microencapsulation of probiotics for gastrointestinal delivery. J. Control Release 162(1):56-67.

de Vos P, Faas MM, Spasojevic M Sikkema J (2010). Encapsulation for preservation of functionality and targeted delivery of bioactive food components. Int Dairy J 20(4):292-302.

Ding WK Shah NP (2009). Effect of various encapsulating materials on the stability of probiotic bacteria. J Food Sci 74(2):M100-M107.

Fang Z, Bhandari B, Garti N McClements DJ, (2012). 73-109 4 - Spray drying, freeze drying and related processes for food ingredient and nutraceutical encapsulation, Encapsulation Technologies and Delivery Systems for Food Ingredients and Nutraceuticals. Woodhead Publishing.

FAO/WHO (2002). Guidelines for the evaluation of probiotics in food. Food and Agriculture Organization of United Nations and World Health Organization Working Group report.London, Onatario.

Fuller BJ (2004). Cryoprotectants: the essential antifreezes to protect life in the frozen state. CryoLetters 25(6):375-388.

Krasaekoopt W Watcharapoka S (2014). Effect of addition of inulin and galactooligosaccharide on the survival of microencapsulated probiotics in alginate beads coated with chitosan in simulated digestive system, yogurt and fruit juice. LWT- Food Sci Technol 57(2):761-766.

Lee Y-K Salminen S (1995). The coming of age of probiotics. Trends in Food Science & Technol 6(7):241-245.

Makinen K, Berger B, Bel-Rhlid R Ananta E (2012). Science and technology for the mastership of probiotic applications in food products. J Biotechnol 162(4):356-365.

Manojlovic V, Nedovic V, Kailasapathy K Zuidam NJ, (2010). Encapsulation of Probiotics for use in Food Products. Springer, London.

Martin-Dejardin F, Ebel B, Lemetais G, Nguyen Thi Minh H, Gervais P, Cachon R Chambin O (2013). A way to follow the viability of encapsulated Bifidobacterium bifidum subjected to a freeze-drying process in order to target the colon: Interest of flow cytometry. E J Pharm Sci 49(2):166-174.

Nedovic V, Kalusevic A, Manojlovic V, Levic S Bugarski B (2011). An overview of encapsulation technologies for food applications. Procedia Food Science 1(0):1806-1815.

Pop OL, Brandau T, Vodnar DC Socaciu C (2012). Study of Bifidobacterium Lactic 300b Survival during Encapsulation, Coating and Freeze Drying Process and the Release in Alkaline Media. Bulletin of the University of Agricultural Sciences & Veterinary 69(2):372-379.

Rokka S Rantamäki P (2010). Protecting probiotic bacteria by microencapsulation: challenges for industrial applications. Eur Food Res Technol 231(1):1-12.

Sandoval-Castilla O, Lobato-Calleros C, Garcia-Galindo HS, Alvarez-Ramirez J Vernon-Carter EJ (2010). Textural properties of alginate-pectin beads and survivability of entrapped Lb. casei in simulated gastrointestinal conditions and in yoghurt. Food Res Int 43(1):111-117.

Sathyabama S, Ranjith kumar M, Bruntha devi P, Vijayabharathi R Brindha priyadharisini V (2014). Co-encapsulation of probiotics with prebiotics on alginate matrix and its effect on viability in simulated gastric environment. LWT- Food Sci Technol 57(1):419-425.

Shahidi F Han XQ (1993). Encapsulation of food ingredients. Crit Rev Food Sci Nutr 33(6):501-547.

Simpson PJ, Stanton C, Fitzgerald GF Ross RP (2005). Intrinsic tolerance of Bifidobacterium species to heat and oxygen and survival following spray drying and storage. J Appl Microbiol 99(3):493-501.

Song H, Yu W, Liu X Ma X (2014). Improved probiotic viability in stress environments with post-culture of alginate-chitosan microencapsulated low density cells. Carbohydr Polym 108(0):10-16.

Teitelbaum JE Walker WA (2002). Nutritional impact of pre- and probiotics as protective gastrointestinal organisms. Ann Rev Nutr (22):107-138.

Tripathi MK Giri SK (2014). Probiotic functional foods: Survival of probiotics during processing and storage. J Funct Food 9(0):225-241.

DOI: http://dx.doi.org/10.15835/buasvmcn-fst:10993


  • There are currently no refbacks.

University of Agricultural Sciences and Veterinary Medicine
3-5 Manastur St., 400372 Cluj-Napoca
Tel: +40-264-596.384 | Fax: +40-264-593.792