 Great, thank you very much for coming to my talk, wherever you are in the world. And I'm talking to you today about structuring oils by enzymatic acerolysis, which is a recent development we have done in our lab. So a little less scattering oriented, even though there's a little scattering for the scattering heads among you. But before I do that, because you're so focused on this kind of thing, let me tell you a little bit about what we're doing in our lab with scattering, since scattering is so important for Lund and for the rest of the gang in here. So just three quick projects. We're doing a pre-nucleation structure of liquid triglycerides in the melts. The whole idea, again, I know it's again, again, again. Whether the triglycerides in the liquid state has some sort of structure before they become solids. We're using sand sacks and atomic scale molecular dynamics. We also were crazy enough to grab a whole bunch of cheese. We were doing some milk coagulation work you may have seen published recently. We're trying. And we also put cheese in a U-sands at NIST and see what we get, whether we can tell a good Gouda from a old cheddar from the neutron scattering profiles. I don't know if I have an answer for that. And the other one that got me very excited is this little machine that you see on the right. That's an U-sax machine, ultra small angle X-ray scattering that is inexpensive Bench-top. It has a Germanium 220 monochromator. It's a Bond's heart camera, as you can see there. And you can go down to cues of maybe 10 to the minus 3, X from to the negative 1. And it plugs to the wall 110 volts and it has an air cooler. Would you believe that for a U-sax machine? Anyways, hopefully we'll get something useful out of this thing and it won't be a waste. Quick things about the liquid structure triglycerides, a story that I told you on the left hand side here. You can see a tripolmitin triglyceride and in collaboration with Giuseppe Milano from University of Naples. He went from Japan to Naples back. He's an expert in ASNB, one of the best I've seen and a pleasure to collaborate with. It's a project with Gianfranco Mazanti, Derek Russo, Amarangoni and Giuseppe Milano. And he had some simulations of tripolmitin crystallizing, as you can see from an isotropic melt into some sort of lamellar crystal. And then we told him, hey, can you look really early? And of course, with the judicious use of some coarse-graining, these are the glycerols. And we discovered that it's actually the glycerols agglomerating that have a role to play in the structure formation prior to forming the lamella. Very interesting stuff. And so combine that with some very good work done by Gianfranco at NIST, in which we of course scattered the neutrons off this still-not-crystalline material. And look at this. This is for you, an artistic rendition of clusters. Of clusters of the blue ones are the glycerol groups, these clusters of between five and eleven molecules per cluster prior for it to be coming lamella, showing it's not There was another group, of course, from Marjorie and at Al, who showed that by using X-ray scattering that this was the case. We actually see clusters. We do not see lamella. So really cool. Hopefully we'll get this published soon. And this is the cheese stored. Would you believe that those are different cheeses in the different ranges? We're trying to figure out what's going on with the different power slopes. These structures are a little bit complex to understand. So hopefully one day we'll be able to tell you what a good old cheddar is versus a younger cheddar. But going back to the task at hand, let me talk to you very briefly about enzymatic glycerolysis of oils for structuring it. Let's start from the very beginning. You know that they're solid fats and liquid oils. They're both trisor glycerol molecules, glycerol with three fatty acids. Of course, the melting point of solid fats is higher. So they're solid at room temperature. You see here coconut oil, cocoa butter, and this I guess is palm oil or maybe palm kernel oil. You see the little palm fruit with the kernel and the flesh, right? Palm oil comes from the outside and palm kernel from the inside, from the seat. Now the liquid oils, they have unsaturated, polyunsaturated fatty acids that give the triglycerides a lower melting point. Hence, they're liquid at room temperature. We know this dynamic. Here's an example of solid fats versus liquid oils and their saturated fat content. Just FYI, so that you don't think that a liquid oil is completely unsaturated. Coconut oil 80 to 90% saturated, palm kernel oil 80, cocoa butter about 60%. You would think it'd be higher, right? Palm oil about 50% saturated fat. But look, surprisingly, a little bit, cotton seed oil is 30% saturated fat. Coconut oil, like you call it coal-sized things, 7% saturated fat, where soybean oil, soybean oil is high, 16% saturated fat, and peanut oil and rice bran oils in between. There's a variety of saturated fat contents even in the liquid oils. This saturated fat content leads to crystallization of the material when you're undercooling it, of course, below the melting point or the highest melting triglycerides. You have an actually a profile of solid fat content, the amount of crystalline mass usually determined by NMR as a function of temperature, right? And the unilever where the pioneers in this area of using this parameter to relate to some sort of functionality of the fat. You can think about hardness, plasticity, and also mouth feel if the thing never melts in your mouth and it leaves a coating. So here you have, for example, coconut oil, here you have palm oil, and even with the liquid oils, they're mostly flat and they don't have any solids as a function of temperature above refrigeration temperatures. But some of them do. For example, look here, peanut oil and cottonseed oil, they do reach about 8-9% solids as a function of temperature. And of course, there's all kinds of functionality associated with this. As you know maybe, you mix this solid fat with some liquid oil and you get the predictable decreasing amount of solid fat as a function of temperature. So the more liquid oil you add, the more you lose this solid fat. Now this is a very like thermodynamic view of things, amount of crystalline solid. We've done a lot of work in the area of crystal structure, and this doesn't take into consideration crystal structure whatsoever. You can have the same amount of crystalline mass and have a large crystal or a small crystal and you get completely different rheological and or functional properties. But that's a story, I guess, from years past, even though we continue working on that. Now margarine shortening, I mean, you've heard a lot about trans-fat-free. People do not want hydrogenated fats. So we all went in 2006 and replaced all the hydrogenated fat and partial hydrogenated fat with palm oil, ended up with food products with a ton of saturated fatty acids, which may have deleterious effects towards cardiovascular health if consumed in very large amounts. And so is there like a next step? How can we then, if we do not want to have so much saturated fat and we do not want to have palm oil, which has become a driver these days, what do we use? Well, and let me just add another aspect to that. Now we have this whole area of the vegan food products, the plant-based foods. For example, you have three examples here of some common brands, there's a lot of these going on. If you want to add fat to these things, do we add a lot of palm oil to it? And then we have issues with sustainability. But do we want to put a lot of saturated fat into these allegedly healthy food products that go to fat for all of these guys is coconut oil, maybe mixed with a little bit of liquid oil. Everybody uses coconut oil because they want to avoid having the sustainability issue of palm oil. And it's the only solution right now and it's cheaper than cocoa butter or shade butter, which could be another option for these things. And so let's talk about health and sustainability a little bit, because this is driving a lot of the research that we do in our lab right now. We talked about saturated fat, so you're going to put coconut oil in it, or maybe you put palm oil in it. But at the end of the day, what have we learned from saturated fat? Excessive consumption of saturated fat will raise your bad cholesterol, low-density lipoprotein in your blood serum. It will also raise your total cholesterol, but it also raises your good cholesterol. So when you take the ratio of good to bad cholesterol, it's fairly neutral in that respect. Now, the monounsaturated fatty acids do not really affect the bad or total cholesterol. They raise some of your good cholesterol. So this negative trend from the monounsaturated fatty acids is considered to be beneficial in terms of a decreased risk of getting cardiovascular artery disease. That is, and if you replace the carbohydrates with fat at isocaloric levels. One man's thing told me that you have to qualify these things. You're replacing carbs with fat or fat with carbs. If you replace it with protein, you may have a different story, but this is how this is done. And the polys were always good. Polyunsaturated, linoleic, linoleic decreases your bad cholesterol, decreases your total cholesterol, increases your good cholesterol, and this decrease in this ratio of good to bad is, sorry, bad to good, is good, is even further. So polys have always shown from 1957 with quiesce's work, this has not changed. And this is what men's thing finally wrote for the World Health Organization in 2016. There's really no issues with this concept that have never been. The only issue here is that saturated fatty acids, when you take the ratio, are fairly neutral. So are they bad? The question is, if you have high cholesterol above 200, I would say milligrams per deciliter in your blood, you do not want to eat anything that raises your cholesterol. Because you're outside the range where you should be concerned about this ratio. You should be concerned about your total cholesterol levels as well. If you're within a normal range, sure, let's talk about this. So this is where the saturated fatty acid story comes with health. You know, nutrition is always complicated. Now what about sustainability? We spend a lot of time thinking about sustainability these days. We do not want to do in our lab research any more that doesn't contribute somehow to trying to prevent some of these things happening in the world. We know I come from Latin America. You know that the whole Amazon is being cut down to put pastures for beef. I mean, I like beef. I eat meat, but that's ridiculous. We cannot lose the Amazon to like us because we're going to eat steaks. We just can't do that anymore. I mean, we can't do it, but we won't be here in 100 years. And so the last rainforest, which are the cradle of biodiversity, the Amazon rainforest, the Congo Basin, some of the Southeast Asian forests, you've heard about Borneo, that there's very little left of this. I don't think there'll be much of the Amazon left in the next 20 years. And so this is how, for example, Malaysian Indonesia looked. There used to be a lush forest and let's not be like stand on soapboxes here. In North America it was done, in Europe it was done. The native forests were cut and they put in whatever. But right now what is happening in the tropical world, this is happening right now and that's where biodiversity is and it's a bit of a crime. Now, on the other hand, for example, palm oil, even though you're cutting the trees down and putting the palm plantation, it is the most productive plant. So per hectare, it produces, look at that, 3.3 tons of oil per hectare versus any of the other guys, which are, for example, soybean oil, 0.4 tons per hectare. So you need a lot more land when you're growing soybean for oil than for palm oil. So that's on the posited side. On the negative side of palm oil here, there's a complex graph, okay, is species threatened per million hectares of palm plantations. And it's about 17. So look at how negative the effect of having a palm oil plantation is on biodiversity for the threat to species. Now look at coconut oil, it ain't that much better, I mean, yeah, it is three times better. But it's still quite a bit worse than, for example, olive oil and some of the peanut oils and some of the other ones around. So because the tropical rainforests are the cradle of biodiversity, if you put anything in it, a road, a building, a palm oil plantation, you're going to be threatening a lot of species. So this is at the core of the problem of palm oil and why Europeans particularly have been very active saying, no, we don't want any palm oil. Whether that's right or wrong, that's not my place to say, I'm just pointing out to the fact that it has a strong effect on biodiversity, even though it is the most productive plant for that. So you would think it'd be good, right? So we don't want to have saturated fat because of the health issue. Let's say we said that palm oil is a bit on the wrong side of the fence here. We have done a lot of work in the area of fat nematics and that's still a very, very important area these days in which we take liquid oil and we structure it using self-assembly mostly and we explore using the techniques we love to use. So I'm not going to talk to you about this either, but this is a review we did in Soft Matter recently and you can have direct oil structuring. So you put in the liquid oil, the magic sauce, and it could be a polymer like ethylcellulose or maybe chitin. This is a network of ethylcellulose polymer that traps oil within. That's sort of a physical entrapment within something that looks very much like a hydrogel, the ethylcellulose oleogel. Or you can add these magic ingredients. You've probably heard about the hydroxylated fatty acids. For example, brincinolate acid or hydroxysteric acid. They form these fibrous structures, some of them, the very famous cytosterol, cytosterol and oryzenol that Aaron bought at Al developed at Unilever again. They have a patent on it, but there's many others, right? So there's fatty acids, partial glycerides, natural waxes are big, fatty alcohols at low levels, let's hope less than 5%. And you get something that looks like a fat, whether it's edible or not, or allowed, that depends on the regulators, whether it's cost-effective, big problem, whether it has the rheological properties, another huge problem. These things still need quite a bit of work. Are they natural? That's arguable. They could be. You can have structured by-facing systems. So if you have an emulsion, for example, a gel emulsion or a structured emulsion, we have worked a lot with monoglyceride gels, which are basically monoglyceride mesophases, which trap oil within, which are dispersed within a water medium, and they look like a fat. That's another option. Or you can have indirect oil structuring via solvent exchange, or by using emulsion templates or foam templated, for example, polysaccharides can be used in order to physically trap oil. So lots and lots and lots of things that, so you could think that this could be the fat of those vegan foods. Sustainable, natural, who knows. No, it's a complex question. But what about giving you another option? What about glycerolysis? This is nothing new because glycerolysis was invented a long time ago to make monoglycerides and by-glycerides. What is glycerolysis? You take a triglyceride, you add glycerol, which is a trihydric alcohol. You can add a chemical catalyst or an enzyme. In our case, we used Candidantartica lipase. You can use Mucormehyde lipase. But in the end, what you end up making is, notice how the fatty acid at position three, now it's located here. You have what used to be the glycerol, now you have a monoglyceride, and you're left behind with a diglyceride. Eventually things will go to their equilibrium state. And yes, there's a lot of acyl migration. We have tried one, three specific lipases. They do not look that good, just because of acyl migration issues. That's another question. But look, you end up with a mixture of diglycerides and monoglycerides. Let's not cover this. So what is important about monoglycerides and by-glycerides? Think about this. We have the same fatty acid composition. We still have R1, R2, R3. R1, R2, R3. It has not changed. Hence, you don't have to label it as modified because you haven't changed the molecular composition. Isn't that a loophole or what? And secondly, monoglycerides and diglycerides have higher melting points than triglycerides for the same composition, sometimes over 20 degrees higher. So what have you done? Even though you may have a liquid oil with some saturated fat, you convert it to the partial glycerides, and now you have a fat because you've raised the melting point. And you don't have to tell anybody that you did. So in the first part of our work, we just try to optimize the reaction conditions. We were lucky to get this into nature, into nature food. You know that nature has a new journal, which is part of the actual nature family. And here we did, and they want it. They're very aggressive about this. They wanted to have that long title that we want to replace palm oil and food products. We just wanted to make glyceralysis products, but propaganda is good sometimes. And so we try to optimize the reaction. So notice how in time, this of course is like 72 hours of reaction, but we do that because we had low enzyme loadings. You can optimize this. You can go into a plug flow reactor or semi-batch if you wanted to. Don't focus on that. But look at the conversion of diglycerides, how they change in time. Monoglycerides, how they change in time. And as a function of reaction time, but also the triglyceride to glycerol molar ratio. Always we found that a one-to-one was the way to go in case you're interested in doing this reaction, one-to-one, multi-mol glycerol. And the commercial enzyme, candidate Antarctica, mix, mix, mix, mix. You can do it probably at home. And you obtain the reaction after just a few days. I'm meanable to do in a small scale. Interesting, interesting technology. What else? So here we actually monitored actually the formation of mono-oldian, mono-palmitin, mono-lenolian. That's 18-1, 16-0, the saturated, and the 18-2. Again, as a function of reaction time or glycerol to triglyceride ratio, notice the expected increase as a function of time of the three species as the reaction continues. And interestingly enough, a bit of an optimum, again, at this glycerol to triglyceride ratio of one-to-one. And why not? Of course, we measure the solidified content as a function of time and notice the expected increase both at five degrees and at 20 degrees. I did not tell you, this is cotton seed oil. And again, the glycerol to triglyceride ratio, the solidified content peaks, actually. In the case of five degrees with plateaus, 20 degrees for some reason right now that become obvious in a bit, we actually have a peak. For this, again, showing that the one-to-one ratio is the way to go. And by the way, the reaction was run at 60 degrees Celsius. So at 60 degrees Celsius, the enzyme is at its optimum and you're not overheating the oil and you're keeping it also safe. Notice just a basic practical test on oil loss as the reaction continues and you make the monodiglystrides, the ability of the fact crystal network to trap oil increases. And you have here this plateauing after about 48 hours under the conditions we did. Interestingly, the glycerol to triglyceride ratio at the one-to-one ratio, we had no measurable oil loss based on, we put it in a little centrifuge tube and spin it for about 30 minutes at 10,000 Gs and see how much of it separates up. And this is how we do this at room temperature. So it's harsh conditions. If we do a differential scanning calorimetry on this again as a function of reaction time or glycerol to triglyceride ratio, this is cotton seed oil. Notice this in a differential scanning calorimeter. Notice the onset of crystallization at around five degrees and as the reaction proceeds, we see the growth of a peak associated with the monoglycerides. So you get this high melting peak coming up as the reaction continues and there with the glycerol to triglyceride ratio, interestingly enough, the one-to-one has the arguably a little bit higher crystallization peak as well. So there's definitely an enhancement of the minimal enhancement over the other glycerol ratios at that particular glycerol to triglyceride ratio, which has been corroborates all the other information that we got before. There's nothing fancy schmancy here. In terms of isothermal crystallization, we fit the crystallization curve to an Avrami model. Derived the rate constant and the Avrami index, but combined the two in a half-life or a half-time of crystallization. And notice here how let's just look at one temperature, let's say 10 degrees Celsius. At the particular ratio of one-to-one mole to triglyceride to glycerol ratio, we have the lowest half-time of crystallization. So it goes faster. It all points out to higher melting peak, lower half-time of crystallization, higher rate constant of crystallization. What would we expect at the one-to-one ratio to have more numerous small crystals for the this amount of glyceride, by the way? This is the glycerol is a soil. It makes sense that you have a greater nucleation rate at that particular glycerol to triglyceride ratio and the photograph say you don't need to analyze this. We did with a box-counted fractal dimension and of course the space fill, that's all that number says right now, by the way, I'm not implying fractality, but just the space fill and it's more filled, right? With crystalline mass. So that's good. It's good for actually making industrial fatty products to have a lot of nucleation and not have these gigantic crystals, which then can actually stay separate from the liquid oil. And let's look at some of the optimized conditions for cotton seed oil. Look at the enormous difference between the liquid oil and the cotton seed glycerol as it's brought up, we're up to 30% and it has a very gradual decrease to about 55. That is remarkable. I have not added any saturated fat, any trans fat. I have not hydrogenated. I have not added some weird self-assembled molecule that nobody wants to approve. It's just transform it from the monos and from the triglycerides to the partial glycerides. A bit of a picture of the structure and then here for you some sacks and wax of these things, you actually see the lamellar, the formation of a lamellar structure for the monoglyceride. It's a lamellar structure. So you get one, one-half, one-third, one-fourth. And then the wax scattering pattern is characteristic of a triclinic crystallizer. So it's in the thermodynamically most stable form. So you don't have to worry about meta-stability. You know the one, the beta prime, beta thing they talk about fats, it's not going to a beta prime. It's not going to our therombics, not going to meta-stable. It goes directly to the most stable form, probably because it's diluted in oil. So phase two was to, the project was to go into the why not try all the oils in the universe? That's a bit of an exaggeration, but here we have cotton seed rice bran, tiger nut. I bet you do not know what a tiger nut is. Check it out. Peanut, sesame, soybean, olive, canola oil, high oil lake, canola oil, high oil lake, olive oil. We emptied the shelves in the lab. And here's the saturated fat content of these things from super high to low and your lake acid contents. And let's rip through them, the enzymatic glycerolysis reaction of all of these. The gray lines are the original liquid oil. The black lines are the glycerolysis product. Let's not look at it too care, like too much, but look at how black has a high melting, high melting peak, high melting peak, high melting. They all get high melting peaks. Even oliveing has a nice, now, olive worked fairly well. That should get some people excited about having olive oil spreads. Of course, they have different microstructures. We just put them there for the record. But difficult to, right now, there's not enough time to talk about the relationship between microstructure and properties. And again, from the X-ray scattering patterns, all of them 4.6, 4.6, 4.5, 6, 4.6. They're all in the triclinic crystallized in the triclinic form of the monoglyceride. And you can see here the lamellar structure of the monoglyceride. So it makes a lamellar crystal, all of them related to their average fatty acid composition in the triclinic form. But do not forget, we also have a secondary peak for diglycerides as well. So we actually have monoglycerides and diglyceride crystals present in these mixtures. We'll get back to that in a second. Here are the solid fat content versus temperature profiles for all those oils we looked at. Here's the tiger nut oil, up to 35% solids. Very interesting. It's a grass, by the way, with a, maybe you've seen. Here's the difference between the liquid and the solid. Notice the increase in peanut oil. The very, very interesting increase. And cotton seed oil, we saw that before. Look at rice bran oil goes up. Olive oil went up to 25% in refrigeration temperatures, maybe not solid enough at room temperature for a fancy margarine. High-lake canola oil, soybean oil increased an interesting extent. Maybe we get enough crystals for pickering stabilization under refrigerated conditions for your little dressings, right? And multifire-free stuff. And sesame oils, canola, and the high-lake algal oil didn't go up at all. But notice, potential for very different ones. Maybe once we develop a greater understanding of more molecular detail on these, we can actually design one with the most amount of solids. Also notice cotton seed, how it has a gradual reduction in solidified content as a function of temperature, while peanut and tiger nut have more for two-stage. We were coming to some sort of conclusions about that. What defines this two-stage melting versus the more gradual melting? And here we have a very high-lake acid content, and here we have a low-lake acid content. So the ones with low-lake acid, more gradual. The ones with high-lake acid, more two-stage. And rice bran oil is somewhere in between of the two. Is that completely true? This could be one of those correlations that are not cause and effect. So don't put too much emphasis on that. It's just a preliminary step. So the important points to take from this is that saturated fatty acids contribute to the solidified content of five. It's good to have an oil with high-saturated fatty acid content. Olake acid contributes to SFC at five when saturated fatty acids are present above five percent. So olake acid is good, but you need to have some saturated. And very high-lake acid contents are required to achieve a high level of solids when saturated fat content is less than 10%. As we said, that goes along with the second point. But if you have more olake acid content in your mixture, you have the more abrupt two-stage kind of decrease of SFC as a function of temperature. Now, we also have this. This is complicated to talk in a short talk like this. Molecular interactions between the species present there. And I was just going to point out to a few of these. For example, if you get monopalmitin-16-0 and monosterin, you have ideal solution behavior. You have ideal mixing between the two solid states. We're talking about solid states when it's in the alpha polymorphic form. But in the beta polymorphic form, you have eutectic defense. So you have now a decrease in the melting point. So you have to keep in mind how much CaT monoglyceride versus C16 monoglyceride you have. If you're wondering what on earth this is, this is a subalpha phase, another strange phase that the monoglycerides do. Monoleon and monolinoleon. Well, let's not talk about this little one here. They have fairly ideal mixing behavior. There's mixing in a solid state. Monopalmitin and monoleon, what's the deal with that? Notice here, this is the melting point of monoleon. And you can actually incorporate up to almost 20% monopalmitin in the monoleon crystal. And beyond that, they completely separate and you see two melting peaks. And you have the appearance of this high melting peak. And we believe that the appearance of a high melting peak is positively correlated to enhanced structuring. So you want to be above 20% palmitic acid to get the benefit of having a high melting fat crystal network, a higher melting fat crystal network, stabilizing your system. That's what we learned from these two. So we can actually begin thinking about designing a fad this way. For example, it gets more complicated. Dioleon and monoleon also have very complex mixing behavior. And notice that they're miscible. I mean, as you're adding more and more and more monoleon to dioleon, they form a mixed crystal. And then after beyond a certain point, 50% call it, the monoleon separates out and forms that high melting peak. So we should probably be above a ratio of 50-50 weight by weight of monoleon dioleon. Sorry for not putting in terms of more. And again, you have dioleon monopalmitin. I'm probably completely confusing you here. Again, showing similar solubility at certain levels and then separation of the two species, which is what you want in order to get enhanced structuring. So it's important to understand the interrelationships between monopalmitin, dioleon, and then, of course, a different species. You should know all of them. We have done this with two, with three, and with four components as well. Hopefully this will be published in crystal growth and design soon. At the end, it gets too complicated. But we can learn some rules about it on the way. Very quickly, applications. That's a tiger nut. It's a bit of a grass. And that's a tiger nut seed. And that is a margarine, a commercial margarine made by Script Surface Seed Exchange. Here is a commercial margarine of five degrees. And here is butter, this light line here. And this is our commercial tiger nut glycerolus' margarine. No emulsifiers and no saturated fat, other than the one that used to be in the tiger nut. And here it is. This is a back extrusion, by the way, profile. So when you extrude it into a cylinder, it comes out. And it's a good idea. It gives a good idea of plasticity and of hardness. What about the melting behavior? I think the commercial margarine and the tiger nut margarine look very similar. And solidified content versus temperature profile. A tiger nut crashes a little bit before the commercial margarine, but that may not be bad for a fridge-ready kind of product. The solidified content is relatively low for both at refrigeration temperatures. So maybe if you're in a warm country or maybe a hot day in the Netherlands, you will maybe become a little fluid liquid. But we, you know, in combination with other, maybe a little cotton seed oil, will give you the profile you want. And this is after 10-month storage of the tiger nut margarine. We still see the, you know, the regular structure of a margarine product without too many changes and with the required stability. Meat fat analogs. Here you have 50-50 coconut sunflower in the open triangles. Here you have tiger nut glycerolus' product. I would say it's even more advantageous than the blend. So we're beginning to incorporate these in all the vegan products because I think it could replace coconut oil without being palm oil, without being hydrogenated. And look at this, stabilization of peanut butter. So if we take peanut oil and we do a glycerolus' product, now we have something with salt that doesn't separate out and you don't have to add hydrogenated fat. The only problem is that you have now the oil and the flour. So we actually ran the reaction in ground peanuts and it worked, but it takes a long time. But it wasn't impossible to add glycerol and the enzyme right into the mixture. But you do not have any more separation. The annoying separation of natural peanut butter is obliterated by using the peanut glycerolus' product. So in future work, we need to define the critical molecular species necessary for maximal structuring. A bit of those phase diagrams that I probably will board you to tears with. We're going to start throwing this thing in the USACS machine because we'd like to understand the nanoscale structure of the glycerolus' product like we did for the triglycerides, if you remember our work. Are there nanoplatelets of these things? So what is the nanoscale structure of these? And then relate that to rheology, right? To both South and Laos like we did for the triglycerides as well. And just keep on exploring more applications in all food categories, in relationship to itself, emulsifying properties, pickering, stabilization. And I would like to see how it works in lamination, hence plasticity, but I'm not holding my breath there. That's almost a shangri-la of all fat work. Can you make a good croissant with that? And with that, I thank you very much for your attention. Thank you, Alessandro, for a very interesting talk. There is at least one question here, and I want to know how much free fatty acids is produced in the process. If you get that, do they influence the melting behavior? One percent at the end of the day is the one left. So we try to keep it within... We also did a work in which we removed it. And actually once with the free fatty acid enhanced structuring better. But so you can leave them in at one percent if you want to, or you can do a little bit of physical refining some bleaching, if you want, some bleaching earths. But we didn't find anything negative with having one percent free fatty acids. Okay, thank you. I have a quick one also. So, Alessandro, your glycerol and triglyceride ratio of one that's optimal and so on. I suppose it's a basic mechanism of things have to find each other, so it's more kind of a geometric effect. But then in your slide number 26, wouldn't you then expect the numbers to be symmetric when you have half to one and one to two ratios? If you get my point. Yes, the only thing I have to say about this reaction is that we still do not understand the dynamics of the reaction completely. And remember that we only saw it for a certain amount of time. So there's a lot of change that happens before and afterwards. One could argue that maybe water accumulates and you start getting hydrolysis. So you have competing hydrolysis reactions as well happening. So yes, they should be symmetrical. They should be symmetrical. And you have this acyl migration issue, which is gigantic, right? And so we didn't know we would expect with the other equilibrium, but you could also be impartial. You're not reaching the complete equilibrium for the positional distribution.