Ultrasonic-Enzymatic Diacylglycerol Production

  • Diacylglycerol (DAG) rich oils have shown a high nutritional value since they are digested and metabolized in a way, which significantly reduces body weight.
  • Diacylglycerol can be produced by the hydrolysis of palm oil using a commercial lipase as a catalyst under ultrasonication.
  • By the ultrasonic-enzymatic hydrolysis, DAGs can be produced in high volumes at very low processing costs.

By an ultrasonically assisted bio-catalyzed hydrolysis, standard vegetable oils can be turned into DAG-rich edible oils with a high nutritional value. The ultrasonic-enzymatic hydrolysis provides a good yield of a diacylglycerol-rich oil in shorter reaction times and under mild conditions.

The benefits of the ultrasonic hydrolysis:

  • fine emulsification
  • increased mass transfer
  • mild conditions
  • short process time
  • temperature controlled
  • inline production

Research & Results

Awadallak et al. (2013) have investigated the ultrasonically assisted hydrolysis of palm oil using Lipozyme RM IM as biocatalyst. In the two-step reaction, ultrasound is used to promote the emulsification of oil and water. In the second step, the enzymes are added for the catalytic conversion.
UP200S with glass reactor for the ultrasonically enhanced enzymatic hydrolysis of oilUltrasonic setup as used in the research of Awadallak: UP200S (200W, 24kHz) with glass flow cell (see picture on the left).
The research group found that following two-step process results in the best results: the ultrasound probe was inserted to a depth of about 10mm into the water/oil system, the power was adjusted to 80 W and turned on for 3 min to emulsify the system before being removed, and then the enzyme (1.36 wt.% water + oil mass) was added while the solution was mixed by magnetic stirring (300 rpm).
The ultrasound treatment was carried out using the ultrasonic probe device UP200S.

Thus ultrasonically assisted bio-catalysis yielded DAG oil with 34.17 wt.% concentration after 12h reaction time. The sonication step itself was very short with a duration of just 1.2 min.
Since sonication is applied as first step to obtain a fine-sized ultrasonic emulsion, this two-step process has advantages for large-scale production: Its energy costs are very low and its short emulsification time allows the use of reduced continuous ultrasonic equipment to feed large hydrolysis reactors.

Continuous ultrasonication in an ultrasonic glass reactor (Click to enlarge!)

Ultrasonic inline emulsion

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Ultrasonic Equipment for Pilot & Production

Hielscher Ultrasonics is your long-time experienced partner for ultrasonic processes. We manufacture powerful and reliable ultrasonic systems for any volume. As supplier to the pharmaceutical and food industry, we are specialised in the manufacturing of ultrasonic equipment as well as in the consulting for ultrasonic processes and their implementation into production streams.
The table below gives you an indication of the processing capacity of our ultrasonicators.

Batch VolumeFlow RateRecommended Devices
0.5 to 1.5mLn.a.VialTweeter
1 to 500mL10 to 200mL/minUP100H
10 to 2000mL20 to 400mL/minUP200Ht, UP400St
0.1 to 20L0.2 to 4L/minUIP2000hdT
10 to 100L2 to 10L/minUIP4000
n.a.10 to 100L/minUIP16000
n.a.largercluster of UIP16000

Hielscher’s MultiPhaseCavitator

Hielscher has developed the unique flow cell insert MultiPhaseCavitator MPC48. The insert MPC48 (see picture on the right) is equipped with 48 fine cannulas, through which the second phase is injected as very fine liquid stream with a diameter of 0.3mm to 1.2mm (depending on the cannula size). Since the second phase is injected already very fine-sized directly into the ultrasonic cavitation zone, a micro- or nano-emulsion is produced by sonication. The MPC48 insert is designed for Hielscher flow cell reactors and can be used for batch and continuous processing.
Click here to learn more about the MultiPhaseCavitator!

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FC100L1K-1S with InsertMPC48

Ultrasonic Flow Cell Reactor FC100L1K-1S with InsertMPC48

InsertMPC48 with 48 fine cannulas is ideal for sono-crystallization and sono-precipitation

InsertMPC48 for even better emulsion results


  • Awadallak, Jamal A.; Voll, Fernando; Ribas, Marielen C.; da Silva, Camila da; Filho, Lucio Cardozo; da Silva, Edson A. (2013): Enzymatic catalyzed palm oil hydrolysis under ultrasound irradiation: Diacylglycerol synthesis. Ultrasonics Sonochemistry 20; 2013. 1002-1007.
  • Dhara R.; Dhar P.; Ghosh M. (2013): Dietary effects of diacylglycerol rich mustard oil on lipid profile of normocholesterolemic and hypercholesterolemic rats. Journal of Food Science Technology 50(4); 2013. 678-86.
  • Dhara R.; Dhar P.; Ghosh M. (2012): Dietary effects of pure and diacylglycerol-rich rice bran oil on growth pattern and lipid profile of rats. Journal of Oleo Science 61(7); 2012. 369-75.
  • Goncalves, Karen M.; Sutili, Felipe K.; Leite,Selma G.F.; de Souza, Rodrigo O.M.A.; Ramos Leal, Ivana Correa (2012): Palm oil hydrolysis catalyzed by lipases under ultrasound irradiation – The use of experimental design as a tool for variables evaluation. Ultrasonics Sonochemistry 19; 2012: 232–236.
  • Souza, Rodrigo O. M. A.; Babicz, Ivelize; Leite, Selma G. F.; Antunes, Octavio A. C.: Lipase-Catalyzed Diacylglycerol Production Under Sonochemical Irradiation.

  • About Diacylglycerols

    Diacylglycerols (DAG) are commonly used in different degrees of purity as additives for enhancing the plasticity of fats or as bases for the food, medicine and cosmetic industries. DAG are also used as estranger oils to separate materials from moulds and as an adjuster of fat crystals, precursors for organic synthesis of products such as phospholipids, glycolipids, lipoproteins, pro-drugs such as DAG-conjugated chlorambucil for treatment of lymphoma, (S)-(3,4-dihydroxyphenyl)alanine (LDOPA) for treatment of Parkinson’s disease and many others. More recently, DAG-rich oil has been used as a functional cooking oil, with a content of least 80% of 1,3-DAG.
    Diacylglycerol (DAGs) can be produced by partial hydrolysis, esterification or glycerolysis through chemical or enzymatic catalysis. Sonication has been proven to intensify the enzymatic catalysis of diacylglycerols drastically. The ultrasonic-enzymatic hydrolysis allows for higher yields of high quality DAGs in a very short process time.

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