Preparation of Fecal Capsules.
The procedures on the net are very vague. Some, in my mind, won't work. Loading a OO capsule then placing it in a 000 capsules then coating it with Shellac seems logical but when I tried it, but there were problems.
There is no mention of filtering. How do you do that without adding a solvent like a saline solution. I assume we don't want any extra water or volume?
How do you transfer the stuff into the capsule?
The first thing I tackled was filtering without dilution.
A 100g sample was scooped into a large food strainer and forced through using a very large plastic spoon over a stainless steel pot. The filtered product did not fall into the pot but rather collected as a mass on the underside of the strainer.
This was spooned off with a tea spoon. A large 50ml syringe with plunder removed was held vertically with a clamp. Slowly, 2 tea spoons were used, scraping each other and letting the fecal matter drop into the syringe. Every so often, a 6mm dowel was used to stir and bed in the matter which also reduces air bubbles.
When full, the plunger was carefully inserted letting about a quarter of the product squirt back into the strainer.
Using tweezers, 5 holes were made in a piece of polystyrene foam and a row of 00 capsules were inserted. The syringe nozzel was inserted close to the bottom of the capsule and drawn up slowly as it filled. This takes care of inevitable air bubbles.
Caps were placed on each, then one capsule was withdrawn from the foam and loaded into a 000 capsule without success. The bottom of the 00 capsule was already soft and sticky. When attempting to insert it into the 000 capsule it got stuck to the wall of it. This was remedied by inverting the 00 capsule before insertion. Next step was a wax coating.
About 30g of organic beeswax (ebay) was placed into a 50ml tall beaker (ebay) and suspended in a pot of water heated to about 80 degrees C. The 000 capsule was held with tweezers and dipped more than half way in the wax, withdrawn quickly with a small shake and blown on. It cooled very quickly! The capsule was inverted and the other side was coated.
I tried Shellac but it was useless for one major reason -- time. It take a long time to dry and you have to hold it until it does. If you put it down, it will stick to the bench. It also tends to make the capsule swell.
In a comparison test, capsules coated with 1 coat of shellac and capsules with beeswax were submersed into a hydrochloric acid solution, pH 1 - pH 2. The shellac coated capsules disintegrated within the hour. Putting 3 coats on 15 - 30 capsules would take all day. The beeswax coated double capsule lasted about 3 hours or less.
Next. Using a single 000 capsule with an inner and outer coating.
The water temperature was ramped up to 95 degrees C. The beeswax tempreature was 90. A cue tip was inserted into the wax then with a 000 capsule in one hand, molten wax was painted inside the capsule in 3 or more stages because it cools quickly.
The cap was also coated but only half way. While still warm, the cap was placed on the main body thereby pushing back residue wax and then removed. This requires a bit of force. The wax adds strength to the capsule.
The capsule body was inserted into the polystyrene foam and fill with fecal matter using the syringe. Once loaded and cap replaced, it was quickly dipped in the wax. Even at 90 degrees, It cools so quickly, I doubt that any damage will result.
These single capsules lasted between 5 -7 hours in pH 1 - pH 2 acid before coming apart around the junction. One capsule was still going strong after 10 hours. What if they go straight through you?
Who knows how long they will last when you take into account churning and the peristaltic action of the small intestine. If I burp and it smells like shit, I will know. My first trial will be 7 capsules over 2 days.
One study found on the web suggests;
"Each patient consumed 15 stool capsules from pre-screened, healthy volunteer donors, over a two-day period."
Another says;
"The single treatment requires the patient to down 24-36 capsules in one sitting.
It takes 24 to 34 capsules to fit the bacteria needed for a treatment, and patients down them in one sitting. The pills make their way to the colon and seed it with the normal variety of bacteria."
Caspule perparation.
Coating the inner and outer shell, results in a capsule that tends to go all the way through unscaved. Some break up but not enough to disperse the contents. It would be better to fill an empty capsule then quickly cap and coat the outside in wax then take it immediately. It appears to remain intact in both water and pH 1 solution for many hours.
With movement though, it will probably break up before reaching the intestines. Using wax or shellac and gell capsules is hit and miss, doesn't seem viable to me.
I will go with a procdure I found here;
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4506624/
"Aliquots of 0.4 mL of FMT slurry were dispensed into Size 1 acid-resistant hypromellose capsules (DRCaps, manufactured by Professional Compounding Centers of America, Houston, TX), subsequently placed within Size 0 acid-resistant hypromellose capsules and then nested within Size 00 gelatin Caps (Capsuline, Pompano Beach, FL). Capsules were administered immediately upon filling and capping."
From what I understand, the gel formulation of the HPMC capsule (DRcaps) is claimed to start disintergrating up to 50 minutes in about pH1. With moist contents inside, it will be a lot faster than that.
DRcaps disolve quickly when in contact neutral water. So what's the point of using acid resistant caps when they will contain moisture?
A Size 0 DRcap was filled with water containing a dye. It was placed in a 00 DRcap and then a 000 gel cap and immersed in pH1 HCL acid in a test tube. This was repeated and immersed in neutral distilled water.
After about 30 to 40 minutes, both the 0 and 00 DRcaps appeared to disolve somewhat into the 000 gel cap.
After 60 minutes, the gel cap in water had shed some of it's outer bulk which was pooling on the bottom of the test tube. There was no visible shedding in the acid solution. The cap in the acid solution appeared to be more integral whereas the cap in the neutral water was a little swollen.
After 90 minutes, the water in the neutral test tube had a slight coloured tinge but the cap appeared to be integeral. There was no colour in the acid solution.
After 2 hours there was a faint colour in the acid solution
After 3 hours, the cap in the neutral water tube had come apart. The cap in the acid solution was still integral.
After 5 hours, the cap (top of the gel capsule) had come off. The inner DRcap seemed to be holding it's shape. The acid solution was coloured but the DRcaps still retained the darker coloured dye and was not totally destroyed.
It seems that a gel cap does better in acidic conditions as apposed to neutral water, but why bother getting DRcaps when both will quickly fall apart in water?
A 00 gel cap was filled with coloured water. within a minute, tiny bubbles appeared on the bottom, on the outside of the cap. It burst with pressure and and hit me right in the friggin eye! It was repeated. Same thing. This doesn't happen with a DRcap. It just falls apart.
A 00 gel cap was filled with coloured water, placed in a 000 gel cap and immersed in acid solution. Within 3 minutes, it blew apart. It was repeated. Same result.
A 00 DRcap was filled with coloured water, placed in a 000 cap and immersed in acid solution. It lasted for more than 3 hours.
This was all done in a static enviroment. With churning and movement in your stomach, I find it hard to beleive that you will get 90 - 180 minutes before the caps release but that's how they did it in the published trials, so I will go with it.
To be on the safe side, I will take 30 to 40 caps with Betain HCL to insure acidity.