Homemade Insulin: Part I

I remember, of course, the day it happened.

He was such a young tike, too.  Not even a year old.  He’d had a fever for the last couple of days, a cough, some diarrhea.  It had gone on like any other cold a kid gets. Something was off, though.  He’d become less responsive over the last few hours, then started breathing funny.

My mom called the pediatrician and told him what was going on.  He asked us to put the phone up to his mouth so he could hear him breathing.

More than 20 years later, I still remember the doc yelling.  I could hear him, over the phone, despite being on the other side of the room.  “Take him to Texas Children’s Hospital RIGHT NOW! I’ll call ahead so they know you are coming!”  He then hung up.

Left us a bit stunned, it did. I remember my parents looking at each other a bit confused, then collecting things to take.  The phone rang.

I answered it.  “Have your parents left yet?” “No, they are still getting their stuff together.” “Tell them to hurry the hell up!” And he hung up again.

I didn’t tell them.  They were going as fast as they can go, and mom would have paused what she was doing to apply switch to backside if I repeated word-for-word what the man said.

Off they went. They told me later the doctors struggled to make the diagnosis.  At one point, the doc came out and told them, “He’s dying, and we don’t know why.  We’ll keep working on him until he’s gone.”

Finally someone had the bright idea to treat his elevated blood sugars.  He went from death’s door–and I mean, right at the edge–to ok in 24 hours.  It was DKA.  He was diabetic.

It was not until many years later that I put all the pieces together as to why it took so long to diagnose simple DKA.  My kid brother, unlike most diabetic patients, goes into DKA at a relatively low blood sugar–somewhere in the 180 range, instead of the 400-1000 range that send most diabetics over the edge.

Fast forward about 3 years. I’ve come home from college for the weekend.  I walk into a quiet house, drop my stuff, and head for the kitchen (I know, typical college kid).  There at the table is my little brother, with blood running down all his fingers and dripping on the table.  Shocked for a second, I recovered enough to yell “What happened?!”.  He turned those blue watery eyes towards me, and, lip quivering, he said, “I just wanted to be a big boy and check my own blood sugars!” Of course I ran to him, snatched him up and held him tight; both of us cried–he from broken fingers, me from a broken heart.

He had the coordination to poke his finger with the lancets, but lacked the coordination to get the drop of blood on the meter’s stick.  So he had stuck the next finger, then the next, until all were running blood. I’m not sure if he was crying from frustration or pain; by that time his fingertips had lost most of their sensation. It was a devastating day for me, however, one I’ve never forgotten, and one that pushed me into the field I work in today.

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So as you can see, I’ve more than a passing interest in diabetes.  My kid brother (who, genius that he is, will almost certainly make his first million before he’s thirty) means the world to me.  I’m highly motivated to find something to help him, in the ever-more-likely event that the grid goes down.  This series of posts are for him, and for the mom that has worried over him since the day he was born.  Ya’ll are welcome to eavesdrop, but please remember, it’s not really for you.

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I have seen the stories of Eva Saxl and her husband as well; so it is possible. Here is the link to the book they reportedly used, if you read German.

Here is the youtube video describing the same thing:

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I start by saying that the truly modern methods of extraction of insulin from genetically engineered bacteria or yeast will not be possible in a grid down scenario.  Not because you filthy rich preppers out there may not be able to come up with the equipment, but because the companies that make insulin are not going to give out samples of the bioengineered critters.  We are stuck with methods of extracting animal insulin from the pancreas of “volunteer” animals.  Please note that in order of preference, you use pig, cow, sheep/goat, and then any other mammal you can get.  Dogs, cats, camels, foxes, whatever. Failing that, fish also have a form of insulin, albeit not much, and not very potent in humans.

If you are in the business, please engineer a new yeast using the new thermostable insulin, then have your brainy folks set up a system that would fit in the back of a pickup truck.  Then send me one.  And one to my little brother.  And ship a bunch to the Third World too. Say, doesn’t the Gates Foundation support things like that?

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Grid down production of insulin is essentially a review of the history of insulin production.  Read an excellent but non-technical summary here, it will give you the background for the following.

No, really, go read it.  It’s not technical, it’s an easy read, and following the step-by-step nature of the progress they made is important.  While you are at it, chew on a bit of true history and read the 1923 Nobel Prize acceptance speech by one of the co-discoverers of insulin, Frederick Banting, here.

Also, you need to know what is meant by a “polar” vs “non polar” molecules.  It’s a fundamental concept to understand; hopefully if you really grok this concept, you will be able to chemically improvise if you don’t have the reagents from the protocols. Watch this:

For our technical review, let’s start with a summary presentation given to the Royal Society of Canada by Collip, found here in archive.org.  This is a sequential description of the various methods they used to extract insulin, from the easiest to the hardest. Editor’s notes [in brackets].

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“In two previous papers a brief outline of the preparation of pancreatic extracts has been given…The main problem in the preparation was to get rid of or avoid the presence of proteolytic enzymes.”

“The first extract used was obtained by ligating the pancreatic ducts of the dog, and waiting from seven to ten weeks for degeneration of the acinar tissue [This removed most of the contaminating proteins and destructive digestive enzymes from the preparation]. The remnant, which contained healthy insular tissue, was removed and macerated in ice-cold Ringer solution [basically saltwater]. By this procedure a non-toxic extract which markedly reduced the blood sugar and the excretion of sugar in diabetic dogs was obtained in small quantity.”

[This represents the simplest method of insulin extraction; basically you make ‘tea’ out of the pancreas by mixing it with Ringer’s solution.  The insulin (which is a charged protein made of amino acids) dissolves in the polar water, and a host of undesirable substances cannot.  You then filter out the insoluble, solid material.  In this case, they poured the mix over several layers of cheesecloth.  The water (with insulin dissolved in it) passed through, and the junk was caught in the filter.  Using this simple approach is very easy and can be done almost anywhere, but problems arise in yield of insulin per kg pancreas, and contamination, problems that persist throughout the history of insulin extraction methods.]

“[The next iteration of insulin extraction used fetal calf]…The foetal calf extract was at first made by macerating pancreas of foetal calves of under four months development [at this point in the calf development, the pancreas has not yet produced the destructive digestive enzymes that reduce the yield] in Ringer’s solution and filtering. Later, 95 per cent alcohol was used in place of Ringer’s solution [this is a tincture of pancreas, instead of pancreas tea]. The alcoholic filtrate was evaporated to dryness in a warm air current and the resin-like residue redissolved in saline. This solution when injected subcutaneously or intravenously into a diabetic dog [but they had to inject a lot, around 10 ccs] caused a marked fall in blood sugar and in sugar excreted in the urine. It was further found that this extract did not contain trypsin, that it was destroyed by boiling, that the active principal was insoluble in 95 per cent alcohol [All of these argue the the ‘active principle’ is a protein, which was not yet established at the time.] and that daily injections enabled a totally depancreatized dog to live a much longer time (70 days) than has hitherto been recorded after such an operation.”

“Potent extracts of the whole gland of the adult ox were obtained in a similar manner, using equal volumes of 95 per cent alcohol and pancreas, with the exception that the alcohol was made 0.2 per cent acid by the addition of HCl. [Presumed to be concentrated HCl] It was found that the fatty substances in the extract could be removed by washing twice with toluol without deterioration of the potency of the extract. The alcohol could also be removed by distillation in vacuo at low temperature and it was found by reducing the volume to one-fifth instead of to dryness that a watery extract of the active principal was obtained. This could be sterilized by passing it through a Berkfeld filter. The extract in this form was given to a human diabetic and results in every way comparable to those obtained on the depancreatized dog were observed. However, owing to the high percentage of protein also present, sterile abcesses formed in a few instances at the site of injection [Still the issues of contamination and purification.].

The Preparation of the Extracts as used in the first Clinical Cases:

“To a small volume of 95 per cent, ethyl alcohol freshly minced pancreas was added in equal amount. The mixture was allowed to stand for a few hours with occasional shaking. It was then strained through cheese cloth and the liquid portion at once filtered. The filtrate was treated with two volumes of 95 per cent, ethyl alcohol. It was found by this treatment that the major part of the protein was removed while the active principle remained in alcoholic solution. After allowing some hours for the protein precipitation to be effected the mixture was filtered and the filtrate concentrated to small bulk by distillation in vacuo at a low temperature (18° to 30°C). The lipoid substances were then removed by twice extracting with sulphuric ether in a separating funnel and the watery solution returned to the vacuum still, where it was further concentrated till it was of a pasty consistency. 80 per cent ethyl alcohol was then added and the mixture centrifuged. After centrifuging, four distinct layers were manifested in the tube. The uppermost was perfectly clear and consisted of alcohol holding all the active principle in solution. Below this, in order, were a fiocculent layer of protein, a second clear watery layer saturated with salt and a lowermost layer consisting of crystals of salt. The alcohol layer was removed by means of a pipette and was at once delivered into several volumes of 95 per cent, alcohol, or better, of absolute alcohol. It was found that this final treatment with alcohol of high grade caused the precipitation of the active principle along with adherent substances. Some hours after this final precipitation the precipitate was caught on a Buchner funnel, dissolved in distilled water and then concentrated to the desired degree by use of the vacuum still. It was then passed through a Berkfeld filter, sterility tests made and the final product delivered to the clinic.”

See how well it worked:  Banting, Best, et al: Pancreatic extracts in the treatment of DM

Next post, we will visit the process of isoelectric precipitation and move forward in time to crystallization of insulin.

 

UPDATE: added YouTube video discussing polar vs nonpolar

6 responses to “Homemade Insulin: Part I

  1. Pingback: Hogwarts: Homemade Insulin – Part I | Western Rifle Shooters Association·

  2. I have type II diabetes. I am prescribed Metformin, a generic form of Glucophage (in Greek it means “sugar eatter”) On Wikipedia history “The biguanide class of antidiabetic drugs, which also includes the withdrawn agents phenformin and buformin, originates from the French lilac or goat’s rue (Galega officinalis), a plant used in folk medicine for several centuries.[110]”
    Worth looking into…

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  3. Quick note on insulin thermostablity: commercial insulins are generally more stable and hearty than the label suggests (i.e. 30 days). A lot of that is probably do to caution on the pharmaceutical company side.

    Doctors Without Borders/U Geneva commissioned a study to examine the loss of potency (which I hope is published soon). Though heat will denature insulin it has to be pretty intense for long periods of time. Sunlight and shaking also destroy potency. However in most cases you can get months of potency even if it is kept outside of a fridge. What will quickly destroy it though it going the other way – freezing. So be careful not to store directly on ice packs.

    You can also look at the ‘Clay Pot’ Olympics study that examined ingenious ways insulin is being stored in Africa. Traditional non-electric evaporation based devices, e.g. Zeers, are more than sufficient to cool insulin in Sahara strength heat.

    Claypot – http://medialibrary.ispad.cyim.com/mediatheque/media.aspx?mediaId=11269&channel=9857

    Pot-in-Pot Refrigeration – https://en.wikipedia.org/wiki/Pot-in-pot_refrigerator

    Thanks for the well researched article.

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  4. Intriguing, but where do I find the later posts? …i.e., “Next post, we will visit the process of isoelectric precipitation and move forward in time to crystallization of insulin.” A little background: having had type 1 for over 50 years, I am always concerned about a ready supply of insulin–especially now since big pharma has increased the price of insulin 10-fold!! :-O BTW, in a “previous life” I was a Molecular Biologist, not to mention I also grew up on a pig farm, so all these techniques described are second nature and now my “MacGyver tendencies” are working overtime! Lol 😉

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