Thionin test , Bee pollen with Naturally New Zealand

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TESTING THE BIOAVAILABILITY OF PLANT CELLULAR MATERIALS

The Thionin Test

What are plant cellular materials?
These are plant foods or nutraceuticals that are cellular in nature and which have intact cell walls. Common vegetables, herbs, Spirulina and Chlorella are examples. They consist of cytoplasm, which contains all the "good" in the cell, surrounded by indigestible cell wall.

What's the problem?
The cytoplasm contains the material of nutritional or neutraceutical benefit. This is usually easily digested provided the digestive enzymes of the body's digestive system can access it. However, the valuable cytoplasm in plant cells is usually surrounded by an indigestible cell wall consisting of cellulose and lignin. The human digestive system does not have either cellulose or lignase, which are the enzymes that can digest cellulose and lignin, so our digestive system cannot digest and penetrate the cell wall to access the cytoplasm. The good nutritional material in the plant cell is locked away within an impenetrable box. The box is the cell wall made of indigestible wood~like material.

Does that mean we cannot digest any plant cellular material?
No. Many plants have soft cells that are easy to disrupt. Ripe fruits are an example. Others have starchy cytoplasm, and we use a clever trick to make these digestible. We cook them. On cooking, the starch grains in the cytoplasm expand so much that the cell wall box can no longer contain them, and the cell wall bursts - its integrity is destroyed. Boiled potatoes become soft and mushy as the integrity of their cell walls is destroyed by the expanding starch. When the cell wall's integrity is destroyed, the digestive enzymes of the human digestive system can gain access to the cytoplasm and, of course, the starch. The indigestible wood-like cell walls remain undigested and make up part of the fiber requirements in human nutrition.

Does this mean a lot of plant cellular material can It be digested?
Effectively, yes. Those plant cells with rigid cell walls and with a non-starchy cytoplasm have poor digestibility. Grass, for instance, has thick cell walls and non-starchy cytoplasm. People cannot survive on grass, at least not until now! Cows and sheep have special means for dealing with this problem, and these adaptations enable them to thrive on grass. They have excellent chewing mechanisms, which grind their foods very finely, and they have microorganisms in their stomachs to digest the cell walls for them, since the microorganisms do have cellulases and lignases. We do not have these microorganisms in our stomachs, so material like grass is indigestible to us. Alternatively, we can say grass, and foods like grass, have low bioavailability. Pollen is another example of a food with low digestibility.

What is "bioavailability "?
This has been a some what loose term used to indicate how digestible the material is. I prefer a tighter definition: bioavailability is the proportion of plant cells in a sample that have broken or cracked cell walls such that the cytoplasm they contain is exposed to digestive enzymes.
I prefer this definition since it gives us a simple way of measuring bioavailability. All we have to do is count the number of cells with damaged and cracked open cell walls. If we then have some way of visualising the cell so that we can distinguish intact cell walls from cracked ones then we can measure bioavailability. My thionin test allows this to be done easily, cheaply and quickly.

How does thionin help measure bioavailability?
Thionin is a large molecule that stains proteins blue. Cell walls contain little protein, so they do not stain very much. In cells with intact cell walls the thionin is too large to penetrate into the cytoplasm, so in intact cells the cytoplasm does not stain, even though it is full of potentially stainable protein. So, cells with intact walls do not stain - they stay their natural color. The thionin is mimicking the large digestive enzymes. They, like thionin, are too large to pass through the cell wall.

But in cells with a disrupted, broken or cracked cell wall, the thionin can get into the cytoplasm. These cells with disrupted cell wall are more bioavailable as indicated by the fact that the thionin can get into the cytoplasm where it stains the cytoplasmic protein a blue or blue-green color. With the thionin test the bioavailable cells stain blue. Non-bioavailable cells do not stain. Enzymes, too, can penetrate disrupted cell walls and digest the cytoplasm. The importance of this test is that enzymes, because they are large molecules, behave just like thionin: they cannot get access to the cytoplasm unless the continuity of the cell wall is disrupted. So thionin is a mimic of digestive enzyme behaviour, and hence indicates bioavailability.

How do you conduct the thionin test?
The thionin test is very easy to do. You can obtain thionin from chemical or histological supply houses. Prepare a very weak solution of thionin in distilled water. A concentration of 0.05% is fine. Place a drop of the aqueous stain on a microscope slide. Into the drop gently place a small amount of the material under test. Place a coverslip over the preparation, wait a few seconds, then observe under a microscope. Count the number of blue stained cells and unstained cells. Express the number of blue cells as a percentage of the total number of cells counted (blue and unstained summed). This percentage is the percent bioavailability of the material under test.

Naturally New Zealand uses this test in their investigations and quality control of their potentiation technology. This technology increases the bioavailability of plant cellular materials, such as pollen and Spirulina. See the separate pamphlets for a description of this technology.

For further information please contact:

Brian Lewis (President),
email: brian@naturallynewzealand.com
Naturally New Zealand

* Please note that this Naturally New Zealand product is not available in New Zealand or Australia.