mahesh_chavan Posted September 1, 2011 Posted September 1, 2011 Currently I am suffering from a liver disease. I can eat diastase + papain tablets for digestion. But when I was doing my B.Sc. we have put one egg in juice of papaya ( containing ) papain. Within 3-4 hours it was dissolved and amino acids were formed in the juice. But how to separate amino acids from the juice ? ( I do not want individual amino acids just a mixture will do ) But papain must be separated for re-use In fact I want to separate carbohydrates and proteins from a food sample. Take an example of soy flour which contains all required amino acids. If I press the beans, oil can be extracted. remaining cake will contain mixture of carbohydrates and proteins. If I use fungal diastase it will convert carbohydrates into sugar ( glucose + fructose) now I want to separate these sugars from protein part and then put this protein part in papain o get amino acids. Again how to separate them from papain . I do not want to use HPLC or other costly equipments. Can I get separate sugars and amino acid using some solvents and doing crystalization ? Again except water any other solvent must be separated and re-used. Please guide how can I achieve this. ( any techniques) Thanking you
John Cuthber Posted September 1, 2011 Posted September 1, 2011 An unorthodox answer that might do is to the ferment the carbohydrates to alcohol and boil that off.
spin-1/2-nuclei Posted September 1, 2011 Posted September 1, 2011 Currently I am suffering from a liver disease. I can eat diastase + papain tablets for digestion. But when I was doing my B.Sc. we have put one egg in juice of papaya ( containing ) papain. Within 3-4 hours it was dissolved and amino acids were formed in the juice. But how to separate amino acids from the juice ? ( I do not want individual amino acids just a mixture will do ) But papain must be separated for re-use In fact I want to separate carbohydrates and proteins from a food sample. Take an example of soy flour which contains all required amino acids. If I press the beans, oil can be extracted. remaining cake will contain mixture of carbohydrates and proteins. If I use fungal diastase it will convert carbohydrates into sugar ( glucose + fructose) now I want to separate these sugars from protein part and then put this protein part in papain o get amino acids. Again how to separate them from papain . I do not want to use HPLC or other costly equipments. Can I get separate sugars and amino acid using some solvents and doing crystalization ? Again except water any other solvent must be separated and re-used. Please guide how can I achieve this. ( any techniques) Thanking you Hello, I am so very sorry to hear about your troubles. I am guessing this will have to be a multi-step separation procedure because you cannot use any toxic solvents. Unfortunately I'm not a food chemist. I believe you will have to start out with a centrifuge of some sort, I have no idea how expensive one of those would be for home use, but I suspect it might be possible to easily send your mixture off to a local lab (small start-up company would be ideal) to have them do this for you and it might be surprisingly cheap - but I honestly don't know. Sorry.. But, if you could add some water to the mixture, the amino acids will stay in that liquid layer at the top and the heavier particles etc will end up at the bottom. Once you have that, it may be possible to separate the amino acids from the solution via recrystalization.. here is a patent on the recrystalization of amino acids. This might be ideal because the citric acid would likely stay in the aq layer after centrifugation as well, and if it did you could utilize that to carry out this procedure: http://www.freepatentsonline.com/5118815.html The problem now is that you'd have to try to neutralize the citric acid with a base - and since I am not a food chemist - I am not exactly sure what would be the best to use for this. Water will not work because the pka of citric acid is approximately 3.5 and the pka of hydronium ion is approximately -1.7 - so the equilibrium would favor citric acid so you wouldn't see neutralization with that. Thus, you need an alkali food that will not f'up your recrystalization.. This is the best I can think of on my own, but I have a friend who is a biochemist and he may have some other suggestions for you. I will ask him to review your post and respond if he can. Sorry I didn't have a more complete answer for you, but please do check back here in a day or so.. if it takes awhile to respond it's because he and/or I am trying to work out a better answer for you which will require some research as I am not a food chemist.. Best of luck.. Cheers
Athiril Posted September 2, 2011 Posted September 2, 2011 How about baking soda/sodium bicarbonate from the supermarket? That is food grade. Also you can bake or heat the powder to form sodium carbonate from that. You can also get food grade sodium hydroxide (not from the supermarket obviously).
spin-1/2-nuclei Posted September 2, 2011 Posted September 2, 2011 (edited) How about baking soda/sodium bicarbonate from the supermarket? That is food grade. Also you can bake or heat the powder to form sodium carbonate from that. You can also get food grade sodium hydroxide (not from the supermarket obviously). Yes, this is a very good suggestion and could possibly work.. However I'd like to caution the OP not to just run with the procedure I've provided until it can be verified by someone who knows something about food chemistry.. The trouble would be not only getting food grade bicarb, but also ensuring that all sugars/etc are removed from the solution prior to attempting recrystalization. Since this person doesn't have access to characterization methods they have to be sure that they aren't getting any other impurity capable of recrystalizing in the final product as well.. It might be necessary to centrifuge - and following that - get rid of sugars by getting amino acids/color and other impurities in one fraction, separate, and then attempt a recrystalization of the amino acids. A food chemist would be able to say if the second step is necessary or if it is safe to simply neutralize an acidic solution containing amino acids and the rest (after centrifugation) - and recrystlize them out - without having other "impurities" commonly found in food also recrystlizing or cocrystalizing. Depending on what reagents are used preservatives, surfactants, etc might cause potential problems... I personally think this person needs a guaranteed (preferably known) fool proof procedure to pull this off, without the use of toxic chemicals, and assurance that no unwanted "impurities" will end up surviving the process and further aggravate their condition if they unknowingly consume them.. Cheers. Edited September 2, 2011 by spin-1/2-nuclei
Athiril Posted September 3, 2011 Posted September 3, 2011 I'm also not a food chemist, but couldn't you test for sugars with sulfuric acid? Also it may be worth hitting up a patent search for all those protein/amino acid supplements - the process they use. Also if you have something like http://www.proteindirect.com.au/?page_id=3 for your country (this stuff here is imported from U.S. iirc.. so you should?) You can get whey protein concentrate and isolate pretty darn cheaply without other crap in it. I don't know if that's helpful but just putting it out there.
Athiril Posted September 3, 2011 Posted September 3, 2011 (edited) double post... Edited September 3, 2011 by Athiril
Athiril Posted September 3, 2011 Posted September 3, 2011 (edited) triple post... sorry one click of my mouse often registers as many clicks with no timing between them.. Edited September 3, 2011 by Athiril
mahesh_chavan Posted September 4, 2011 Author Posted September 4, 2011 triple post... sorry one click of my mouse often registers as many clicks with no timing between them.. -Thank you so much fo your replies. Your suggestion for using centrifuge machine is great, because then can use it separate fat from soya milk instead of using de-oiled soya cakes. I have read that patent for crsystalization of amino acids. Your suggestion for using soda by carb or food grade Sodium Hydroxide is good. And offcourse they must be of food grade. But When I read patent, it become clear to me that I can concentrate aq. solution under reduced pressure to obtain amino acid crystals. Even though they contain air bubbles and of small size, they will do. Because I do not need pure crystals of individual amino acid. I can eat a mixture of amino acid. In any case, If I am not using any other chemicals, there is no risk of any poison getting consumed. I shall be adding diastase first to say de-fatted soya milk. That will convert carbohydrates to sugar. Then I shall use centrifuge to to separate that layer after filtration. if I boil that layer diastase will become inactive. Because diastase is cheap they do not try to reclaim it when corn syrup is prepared from corn starch. Simmlarly, I shall be adding papain to the protein part. And then concentrate under reduced pressure to obtain amino acid crystals. Even if papain gets crystalize, I can eat that. Because papain is present in Papaya fruit and the quantity used will be so minute that it can be consumed without any harm if used in such quantity that it can not further be active ( activity is about 80000 u/g ) Thank you so much. Please let me know if there is any flaw in my proposition. I have come across an article to separate sucrose and glucose /fructose from a mixture using animal charcoal. When the the mixture is passed through animal charcoal column mix. of glucose/fructose comes out straight away. Sucrose is adhered to the column which can be washed using ethanol. I do not have sucrose in the solution. But adsorption chromatography can separate sugars from amino acids. I do not want to use animal charcoal. But will the plant charcoal do ? I do not know which will adhere to the column. Any suggestion / clues for this method ?
spin-1/2-nuclei Posted September 4, 2011 Posted September 4, 2011 - Thank you so much fo your replies. Your suggestion for using centrifuge machine is great, because then can use it separate fat from soya milk instead of using de-oiled soya cakes. I have read that patent for crsystalization of amino acids. Your suggestion for using soda by carb or food grade Sodium Hydroxide is good. And offcourse they must be of food grade. But When I read patent, it become clear to me that I can concentrate aq. solution under reduced pressure to obtain amino acid crystals. Even though they contain air bubbles and of small size, they will do. Because I do not need pure crystals of individual amino acid. I can eat a mixture of amino acid. In any case, If I am not using any other chemicals, there is no risk of any poison getting consumed. I shall be adding diastase first to say de-fatted soya milk. That will convert carbohydrates to sugar. Then I shall use centrifuge to to separate that layer after filtration. if I boil that layer diastase will become inactive. Because diastase is cheap they do not try to reclaim it when corn syrup is prepared from corn starch. Simmlarly, I shall be adding papain to the protein part. And then concentrate under reduced pressure to obtain amino acid crystals. Even if papain gets crystalize, I can eat that. Because papain is present in Papaya fruit and the quantity used will be so minute that it can be consumed without any harm if used in such quantity that it can not further be active ( activity is about 80000 u/g ) Thank you so much. Please let me know if there is any flaw in my proposition. I have come across an article to separate sucrose and glucose /fructose from a mixture using animal charcoal. When the the mixture is passed through animal charcoal column mix. of glucose/fructose comes out straight away. Sucrose is adhered to the column which can be washed using ethanol. I do not have sucrose in the solution. But adsorption chromatography can separate sugars from amino acids. I do not want to use animal charcoal. But will the plant charcoal do ? I do not know which will adhere to the column. Any suggestion / clues for this method ? Hello, I contacted my colleague who is a biochemist and he contacted our other colleague - the food chemist - who is wary of giving advice about something like this online (mostly because it is possible to accidentally concentrate unwanted additives and consume them in unsafe amounts if the incorrect materials and/or procedures are used. I understand your predicament, thus I will see what I can do myself, but I think you should contact a custom chemical process startup company if possible (more on this later) Finding a way to recrystalize the amino acids is not as much of a problem as ensuring there are no impurities. Since you are going to be consuming this I think it would be prudent to triple check whatever procedure you use and if possible come up with a trusted source for all the materials you are planning to use, so that quality control can be ensured. The problem is you don't want to concentrate something that is typically meant to be an additive and slowly dose yourself with unacceptably high levels of said additive ( like preservatives, etc)... Another problem could be unforeseen impurities that end up in their product from packaging process etc that aren't harmful when the food products are being used for the designed purpose, but that over time, and perhaps during atypical use of the product - concentrate and become harmful.. Thus, I think - if you can afford it - you would be well served to take whatever method you decide on and run it by one of those small start-up custom chemical process companies. They may, for a cheap price, be able to help you work out a process that works for you, and provide you with quality suppliers. Many of these places do consulting for breweries, candy manufacturers, supplements, baking goods, you name it... If you contact someone like that, and they can't/won't help you I will do what I can.. but as I said before, unfortunately I am just a computational/green synthetic chemist - so not even in the same neighborhood as a food chemist, but I will do my best to help however I can.. For now I will throughly look over your proposed procedure above when I have time next week, and I will get back to you sometime this coming weekend.. Cheers
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