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Cell lysis with Rotor/Stator Homogenizer?

Birimintingo

Birimintingo

Rising Star
Hi all,

In a few posts, user benzyme stressed the importance of proper cell disruption to make intracellular material available. It took me a while to let that sink in, but it makes so much sense to reach maximum yield.

My question is about the tools; I don't have access to a sonicator and neither the funds. But I have access to a rotor/stator homogenizer. In the thread "Using Lye to break down root bark?" benzyme wrote "rotor-stator homogenization [ ... ] effectively lyse cells in minutes".

Also from my understanding and research a rotor/stator homogenizer is indeed an effective tool for cell lysis, but for some reason this method seems less commonly used and mentioned in texts compared to ultrasound or the french press. But actually rotor/stator cell lysis seems like the most economical and scaleable option out of all, or am I missing something? Specially with a continuous flow chamber you can process large amounts in no time.

Does anyone have experience comparing ultrasound to rotor/stator processed material? Is there perhaps a reason why rotor/stator is not universally recommended for plant cell lysis over sonification?
 
may be an issue with generator tip-size. commonly used rotor-stator generators are 7mm to 10mm, which are large enough to process whatever you can fit into a 15mL to 50mL centrifuge tube. I've used a 7mm to grind up morning glory seeds in methanol, in a 50mL tube.

on the other hand, an ultrasonic cell disruptor probe with a half-inch tip may effectively process anything in a 250 to 500mL beaker/flask, an inch probe tip can easily process a liter.
 
Seems like a lot of money to replace freezing/thawing, acid cooks, and leaving your bark sitting in lye for a day or two. All of which only seem to increase my yeild by the tiniest degree.
 
I think 4 long acid cooks would be more than enough to take out nearly all the spice as the heat lysis the cells. If you are concerned perhaps higher temps like those achieved in a pressure cooker would help. There have been some threads and success stories on here.
 
The CAT scientific homogenizer Unidrive 1000 (but many similar handheld motors are on the market) can process up of 5L per batch with the largest shaft size (30mm diameter). With the addition of a simple flow chamber that can be increased to 2'000 - 5'000L per hour, but I haven't tried that. So that's some serious volume that can be processed, certainly enough for a small scale production of legal plants like Passionflower. Then I would also only pass already finely milled powder through the device, as to maximise lysis.

There doesn't seem to be a whole lot of literature (or I just couldn't find any) about the distribution of intracellular and extracellular alkaloids. The question is whether this homogenisation significantly affects yield. I will make some experiments in the coming weeks comparing yields of homogenised vs. non-homogenised material and report back here. That is really the most important factor that decides whether it's worth the effort.

But the other factor seems to be speed of extraction. I read somewhere that with a sonicator you can extract in 1 minute more alkaloids than in 5 hours of soxhlet. That's quite substantial. So the question is whether also with a rotor/stator it's possible to just run this thing say for 5-10 minutes and skip the common 3x5 hour soaks - saving potentially some 15 hours of time. Definitely will run some tests as soon as time allows.
 
For MHRB cooking or PC is fine, but unless the plant is heat-stable cooking might change the composition significantly.
 
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