Experiment Help, finding a chemical

[ChrisLife]

I've been debating doing a personal experiment for a while now and today decided I'm going to try it even though it might be to difficult to do, or even impossible.

 

I want to keep the Details a secret because well, i don't want to be robbed of my idea. I know it sounds funny, but it has happened to me in the past.

 

Experiment:

 

I have an organism, which can live in mammals(humans included), like a hook worm. It can not reproduce inside the body but only through the stool, but what i'm testing is for a chemical it secretes.

 

The worm secretes this chemical that i believe has beneficial aspects to humans but i need to first somehow grow this worm inside a controlled environment and be able to collect all the chemicals it secretes all while keeping it alive.

 

I would also like help on identifying the chemicals it secretes. If someone could guide me that would be great, as you can see i don't have a background in biology or chemistry, but i was in a biology major for quite a few years, so i have some knowledge.

 

Was thinking along the lines of growing the hook worm in a a controlled environment that mimics a human intestine, which is where i believe they live, and then testing for chemicals, this is were i go blank because i don't know how to test for random unknown chemicals, and even identify them.

 

Going to do some more research right now, but any one that can help me out in anyway would be greatly appreciated!

Once this experiment is done i will of coarse test it on humans, myself, which will be the easier part.

If i can get enough help i will start a thread with the experiment with daily updates on the whole experiment, and my findings.

 

Thanks

Chris

[hypervalent_iodine]

Do you have access to a research laboratory? If not, this is all going to be very hard for you to do in a practical sense.

 

Ignoring that and ignoring the biological aspects, identifying the compounds it secrets will be extremely difficult if not impossible for you to do without a strong background in structure elucidation and organic chemistry as well as access to an NMR and mass spec machine (at the very least). Determining the absolute structure of a compound is often made to look easier than it actually is. It requires a great deal of precise training and prerequisite knowledge to understand the techniques and interpret the data outputs correctly. When you have no clue as to the compound you are looking at, the task is harder again.

 

If you are looking for a specific compound with suspected bioactive profiles and your aim is to isolate, characterise and market it, make sure the next 10 or 20 years are free for you to do work. Truth be told, without the resources, funding and access, achieving this is impractical and most likely not work. If you are not doing this simply for your own benefit, your task will be much simpler though will still prove somewhat unfeasible should you not have the necessary equipment or chemical/biological knowledge at your disposal.

[Horza2002]

If your planning on doing this at home, then heres some advice.....don't bother! It will be impossible.

 

Without access to HPLC, chiral HPLC, high resolution MS, high-field NMR and IR instrucments, you will not be able to work out the structure of the compound. It often takes years to determine the absolute structure of a new natural product.

 

Another major problem is isolating the product in the first place. While a hook worm type creature is relatively simple, it will still produce thousands, if not tens of thousands, of different compounds. What are the benficail effects you think they cause? Because the only way your going to know which of the potential thousand or so compounds you'll isolate, you will have the test them all to see which ones give you the desired effect. Without HPLC, it will be impossible to seperate the mixture. And if your planning on doing this on yourself, which in itself is a very bad idea, you will need lots and lots of each of the compounds.

[farmboy]

How do you know about this chemical but have no idea what it is? This was the plot to a recent episode of Fringe too, you aren't planning to start killing are you lol?

 

And yeah as others have mentioned this is an ambitious project for someone with access to a full lab and analytical services as well as years of training in both biology and chemistry.

 

But seriously, it doesn't even sound like you are sure that this creature exists, let alone that it secretes this chemical. Hope it goes well for you all the same though.

[ChrisLife]

Thanks for the reply's, all help is appreciated!

 

Well, i do know the creature exists in fact there are two which are said to secrete the same chemical. As for the chemical, well i don't even know if it is a chemical but to my best knowledge and some research it seems to be the work of a chemical secreted by this organism.

 

I might have access to a lab, although i don't know all the equipment they have(although i know some of it worked there for 3 years), i have a decent biology and chemistry background, nothing that could probably help me but , i'm motivated. Some of this can be done at home while the chemical part in the lab.

 

I want a little help with a breakdown of the experiment step by step of how i should go about it. I already have a rough idea in my head but its only an idea, lol. I know the biggest problem is isolating the chemical and finding out what it is, but i might get lucky and it might only secrete a few chemicals, couldn't i sent a chemical to a lab for analysis, it might already exist? I have an idea to help it secrete the chemical i want, the chemical is said to be secreted due to the immune system of the host.

 

I feel life you guys may be making this a little more complicated then it is, but maybe your right, i don't know it's an experiment after all, can never know until you try it.

 

Thanks Chris

[hypervalent_iodine]

Thanks for the reply's, all help is appreciated!

 

Well, i do know the creature exists in fact there are two which are said to secrete the same chemical. As for the chemical, well i don't even know if it is a chemical but to my best knowledge and some research it seems to be the work of a chemical secreted by this organism.

 

I might have access to a lab, although i don't know all the equipment they have(although i know some of it worked there for 3 years), i have a decent biology and chemistry background, nothing that could probably help me but , i'm motivated. Some of this can be done at home while the chemical part in the lab.

 

I want a little help with a breakdown of the experiment step by step of how i should go about it. I already have a rough idea in my head but its only an idea, lol. I know the biggest problem is isolating the chemical and finding out what it is, but i might get lucky and it might only secrete a few chemicals, couldn't i sent a chemical to a lab for analysis, it might already exist? I have an idea to help it secrete the chemical i want, the chemical is said to be secreted due to the immune system of the host.

 

I feel life you guys may be making this a little more complicated then it is, but maybe your right, i don't know it's an experiment after all, can never know until you try it.

 

Thanks Chris

 

I don't know about farmboy, but Horza and myself are both PhD level synthetic, organic chemists. I tend to think that we have more than just a bit of an idea with what's involved in structural elucidation. It really isn't an easy task, even for those who have a PhD and have completed post-doc projects in the area. If you don't even know if this 'thing' it secrets is a relatively simple chemical or a protein my first question would be, how will you even know it's what you want (presuming you somehow manage to isolate it)? Isolating compounds can be a painful process as it is, even when you know exactly what you are looking for - which you apparently do not. Chances are, your organism secrets hundreds or more chemicals, all of which you would have to sift through and test individually to find exactly what you are after. Could you send to a lab to get tested? Probably not. I wouldn't think that any labs capable of such a thing would be willing cooperate with single individuals not associated with a research group, industry, etc. I could be wrong on that though.

 

Doing this sort of thing 'at home' is not sufficient if you intend to market your end product. You need a controlled laboratory environment for every step of the process if you don't intend to have your results thrown out and discarded entirely. This takes me back to the point on having access to equipment. If you do not have access to NMR, HPLC, IR, mass spec, etc. etc., you will not be able to do this.

 

As for your organism. You need to ask yourself a few questions. How hard is to to obtain your organism? How much maintenance is required and what sort of equipment do you have to have to be able to cultivate them? We cannot answer these questions without knowing what the organism is, as every organism requires a different environment for optimal growth. You should find specifics in the literature though.

 

I am not saying these things to simply dampen your spirits and nor am I making this sound harder than it is. In fact, that you would think we are doing that tells me you do not have the experience needed to attempt this. In any case, we are trying to be a bit realistic about your idea. It could be a fantastic idea, I don't know, but it is unfeasible for someone with little experience in the field and for someone with no access to the right equipment.

 

Finally, if I may ask you, are you trying to find the active chemical/protein/whatever responsible for the claims that hookworms can eliminate the affects of allergies? Because that's what this sounds like.

 

 

 

I think it's called Helminthic therapy...

[farmboy]

I finished my BSc chemistry last year and I'm currently about half way through an MSc in Chemical Research (which means I have to do about the same amount of lab work as the PhD students in my group, if anything more at the moment as it is an intense 1 year lol) and all I can say to the op, who I think suggested we might be overstating the difficulty involved here, all I can say is that I really wouldn't fancy having to do something like this at my current level even with all the support you get as a PG student in a Uni lab. It will probably take years to get a finished project

 

I mean even just the very most basic things like how do you expect to isolate your compound from the other impurities (that's assuming that you can even get that far. Doesn't sound like you know anything about it's chemical properties, which is (sad but true) essential if you want to get anything like a pure extraction. Do you know about basic stuff like enantiomers and the devestating effect the wrong one can have on a biological system. Theory aside the everyday lab apparatus will be complicated and costly too I think.

 

I don't want it to seem like I am giving you a hard time just for the sake of it, I'd be well happy for you if you successful here, but at the same time I'd hate to see you ruin yourself financially or hurt yourself . Have you considered actualy just going to uni?

[Horza2002]

As Hypervalent has just said, we are both PhD organic chemist...I know from my experince, then even when you know exactly what you've put into a reaction, then working out what you get at the end can sometimes be very difficult. I had one compound that took me three months to work out what it was...and that was using some very fancy and top of the range equipment and it was a very simple molecule compared to natural products. I don't mean to dishearten you about this, but I'm just trying to show you that this will be next to impossible.

 

Yes, you might get lucky and the firsat compound you isolate (ignoring the fact that you won't be able to isolate them without access to HPLC) but I'm sure that won't be the case. There are some labs that will anazlyse samples for you yes, but they will charge you an absolute fortune to do it...they won't be very interested in analysis samples of random people of the general public.

 

In addition to me being a synthetic organic chemist, I am also a microbiologist. I know how difficult it can be growing bacteria to use for feeding experiments and extracting natural products from them. Not only is it very fiddley to grow them in the first place without any contaimination, isolating the natural products takes a long time....and that is even with us overexpressing ther enzymes responsible. I say this to point out the issue that you are going to have growing enough hookworms to be isolate anything useful without contamination. How do you plan on stopping bacteria and fungi competing with the wroms for resource? Do you know which section of their life cycle the worms produce the compound your interested in? Many organisms only produce compounds at a specific time of their life...this period could last on the order of days to minutes!!

 

If we haven't managed to convince you that this is going to be impossibley hard, I will be more than happy to give you any help/advice that I can. I do honestly wish that this would work for you because it could be very interesting to see what happens...I just don't think its possible in a kitchen. Whatever problem you have, just post them on here and I will give you as much help as I can.

[Xittenn]

All difficulties aside if you wish to attain the knowledge required to complete such an extensive task why not first do a study on a similar and already active cycle. I was going to post when you had first opened this thread but I feared the onslaught that might have followed; sorry guys it's like a warzone. It's hard, even more so impossible, regardless nothing is ever learned if the question is not asked.

 

I can't give an example of this with hookworms. I was just discussing a project that I have personally had in mind for some time now in the chat room when you had opened this thread. I wish to ferment Streptomyces Albulus for the production of poly-l-lysine. As such I plan to do a study on the matter as it is a well documented procedure. I would recommend finding the equivalent in hookworms and then you will be in more of a position to think about what it is you are doing.

 

Honestly the equipment is expensive but not ridiculously so. This point is moot if you have the equipment already available, but this isn't even the question so I will refrain from making further comments. I will say though there are a lot of things that people do and this wouldn't be the most ridiculous thing I have ever heard, it still helps to become properly informed before tying up valuable resources; which is kind of what you were doing right?

[mississippichem]

Even if you were to isolate an extract from this worm, you might spend the next couple of years doing bio assays and high-throughput screening to try and isolate the active fraction. Natural products chemistry involves a lot of spectroscopy, a lot of purification techniques, a lot of high-throughput screening, and a lot of failure. It can be incredibly difficult to isolate even lets say one alkaloid from a mixture of twenty something similar alkaloids that you are bound to find from biological sources. Then determining the active fraction can take years even with an interdisciplinary chem/med/bio superteam.

 

The small amount of work I did in natural products was in isolating a vandium miconutrient from these small gross little creatures called tunicates. We never found the active compound (looking for a vanadium based insulin mimic, great idea, difficult to accomplish), which led us to believe that the treasure we sought was in fact a mixture of compounds with a combined mode of action.

 

Such sad stories are par for the course in natural products chemistry. Even if you have a Soxhlet extractor .