Animal Toxins: Snakes

Today, in a special issue of this blog, I'm going to be looking at animal poisons (toxins), specifically snakes. Despite their reputation there are only about 300 poisonous species of snakes, which is about 10% of all known species of snakes. Nevertheless, snakes do possess some of the most deadly toxins, especially cobras, vipers and sea snakes. While snake toxins are not nearly as dangerous or fast acting as some of the toxins found in plants and fungi (think ricin in castor beans, or death cap mushrooms), they can be far more deadly for two main reasons:

   1. While plants and fungi only contain a single type of toxin, snakes often utilise a mixture, making it much more difficult to treat.

   

   2. While plant or mushroom poisons have to be ingested, snake toxins are forcibly injected into your system. 

The likelihood of you dying from a snake bite is pretty much proportional to how good your country's infrastructure is. For example in North America only about 5 people per year die of snake bites, however about in India and Pakistan there are about 50,000 to 70,000 deaths per year due to snake bites.  We can tell that this is not just due to a differing number of poisonous snakes as Australia, the country with the greatest concentration of poisonous snakes has one of the lowest snake death rates.

So why is there such a discrepancy in the snake bite mortality rates. Well as with all medicine speed of response is absolutely essential, and a good infrastructure improves this. Snake bites also require adept medical professionals to treat, as I have mentioned they contain many different types of venom. North American rattlesnakes, for example, utilise at least nineteen different types of toxin. In many cases these toxins can work in a sort of perverse harmony, increasing each others' damage and rapidity of action. 

Toxins have two main ways of killing:

   1. Neurotoxins: As you may have guessed neurotoxins act on the neurons. Neurotoxins are chemical agents that affect the transmission of chemical signals between neurons. Neurotoxins affect this process in a number of different ways. They may block receptors, stopping the flow of transmission, or damaging microtubules or vesicles which carry the chemical messages. This can lead to death by asphyxiation, as chemical communication in the lungs and diaphragm is restricted, obstructing ability to breathe. 

   2.De-clotters: Ok, that 's a made-up word, but I couldn't find the technical term for this. Basically the toxin stops your blood clotting, giving you a sort of impermanent haemophilia, meaning you could actually bleed to death through the bite. The toxin may do this by inhibiting the action of coagulases. These are enzymes that convert fibrinogen into fibrin, which forms the thread of the clots. If these enzymes do not function, the blood will not clot. 

So how do we stop this? Antivenoms are the answer. Antivenoms are actually types of 'two-layer' vaccines. First the animal from which the venom comes has to be milked for its venom. The venom is then diluted to non-lethal levels and injected into an animal (a rabbit for example). The rabbit produces an immune response to the venom. These antibodies are then collected and injected into the person, defending them against the venom. 

The availability of antivenoms and the speed with which they can be received are the main factors affecting how safe you are, although not being around particularly poisonous snake infested areas cant hurt. 

Until next time people!

Gas Chromatography: Method and Applications

I've covered thin-layer chromatography previously in this blog, however chromatography is a big field, and the precision to which it can analyse samples is increasing rapidly, not to mention how widespread the apparatus required to carry it out is becoming. As such I would say it merits a lot of attention as a field of study.

Unlike in thin-layer chromatography, the mobile phase is an inert gas (for example helium), or an unreactive gas (for example nitrogen). The stationary phase is a liquid coating the walls of the column. The stationary phase is chosen to be as close to the polarity of the solute as possible.

The gas chromatograph uses a column (flow through narrow tube). The different constituent chemicals from the analyte are carried up the column by the mobile phase at different rates. The rate is dependant on the chemical and physical properties of the chemicals with the stationary phase. This causes each constituent to exit the column at a different time.

You may be wondering why such complex, and often expensive techniques need to be employed, well fear not I hear your worries and I'm going to illustrate the value of gas chromatography, thin-layer chromatography and mass spectrometry with a couple of case studies.

1. FBI's analysis of EDTA in blood at O.J. Simpson trial:
    Agent Martz and the FBI forensics team used a combination of the aforementioned techniques to detect the presence (or in this case absence) of EDTA in a blood sample from socks and a watch. EDTA is a preservative for blood. If found in high levels it would indicate that the blood samples had been tampered with. The high level of precision, and ability to remove background peaks, of the mass spectrometer meant that this case was where it really established its role as a reliable form of evidence.

2. Analysis of Fire Debris for ignitable liquids:
    In January 2002 there is a fire. One person dies in the fire, and the police decide to bring in a forensics team to investigate. The result is a piece of vital evidence, showing the fire could not have been an accident. The forensics team, using Gas-Liquid Chromatography and Mass Spectrometry find gasoline present on weathered debris. The presence of ignitable liquids (where there should be none) is fairly damning.

Anyway thank you for reading, until next time people.

Dose-Response Relationships

 I'm back, and this time we're going to be learning about the wonderful world of dose-reponse. In the words of John Timbrell: "There are no safe drugs, only safe ways of using them". In a high enough quantity any drug can be dangerous. But how do we assess what this threshold?

One problem is what the real dose is. Different chemicals are absorbed in different ways with different efficiencies, for example cyanide really has to be absorbed through the mouth to be dangerous, however thallium can be potent even when only applied to the skin. It is not only the method of absorption that affects the 'real dose', but also may be metabolised or excreted more or less rapidly.

however well the chemical is absorbed, metabolised or excreted, the higher the external dose, the higher the internal dose. Eventually the internal dose will reach a high enough level that the body's detoxification and excretion processes can not keep up and are overwhelmed, resulting in toxic levels of a chemical. It is particularly important to be familiar with the dangerous dose, as low doses may cause internal damage which is not detectable until it has progressed to the point where it compromises a bodily function.

According to the Paracelsus Principle, as the dose level rises the body cells will show an increasing level of dysfunction and damage up to a maximal effect (function completely inhibited).

As an example consider a drug that lowers blood pressure. The higher the dose of the drug the larger the decrease in blood pressure, up to a fatal decrease. This can be plotted as a sigmoid curve graph.

Another way in which dose-response relationships are represented is if the effect can be measured in a sort of binary way, either present or absent. The number of people displaying the measured effect at each dose is used to plot the graph.This relationship can represent both negative and beneficial effects.

So why does the larger the dose mean the greater the effect? Well for almost every effect in the human body caused by chemical, interaction with some kind of molecule is necessary, for example the receptor on an enzyme. In order for an effect to occur enough of the receptors must be occupied. The higher the concentration of drug, the more receptors are filled by the drug (this is especially important in competitive inhibitors, where they have to have a high concentration of drug in relation to whatever it is competing with). The threshold is where not enough of the receptors are occupied to have an effect. Ultimately this is what you have to think about when assessing the danger of a chemical. Ultimately the lower a dose required to cause the greater an effect the more potent the drug. 

Thanks for reading, over and out!

Ethyline Glycol (A.K.A. Antifreeze)

Antifreeze is a sweeting, but deadly lethal substance. Once used (illegally) for increasing the sweetness of Australian wines. It is technically an alcohol, but while it initially mimics the effects of a glass or two of wine, it quickly turns to something much more serious.

Name: Ethyline Glycol

Formula: C2H6O2

Effect on Victim: Ethylene glycol becomes poisonous when in the body as it is metabolised into several different chemicals. This is caused by the same enzyme that metabolises the alcohol in common alcoholic drinks. The main product is oxalic acid which is poisonous. Also found in rhubarb leaves, oxalic acid causes the pH in the blood to drop (meaning the blood becomes more acidic), inhibiting normal metabolic processes. As if this wasn't bad enough the oxalic acid can also crystallise on the brain and kidneys, resulting in damage. On top of all this oxalic acid reduces calcium, removing it from the body. This produces effects similar to that of tetanus, causing muscles to contract uncontrollably.

Lethal Dose: Has a comparatively high lethal dose compared to other substances we've looked at. of about 100 ml

Diagnosis: Ethylene glycol is a poor choice for a poison as it is relatively easy to spot and takes a comparatively long time to kill. Ideally an analysis of the concentrations of oxalic acid in the blood by gas chromatography would diagnose antifreeze poisoning, however the equipment to perform this is expensive and rare. A much simpler but less accurate diagnosis is analysis of urine, as this will reveal oxalate crystals when ethyl glycol is at deadly levels. Another common test is to take advantage of the fact that fluorescein is often added to antifreeze to help detect radiator leaks. A Wood's Lamp will reveal fluorescence in the patients mouth.

That's it for this rather un-subtle poison. Hopefully now you can see why it's coloured brighter than a poisonous insect. Unfortunately the bright colours and sweet taste mean children are particularly at risk of drinking it. Anyway thanks for reading, see you soon!

Sample Analysis: Readily Made Observations

   Hi everyone, I'm back with a foray into the world of analysis of drugs and/or poisons, so without further ado, let's get started!

   To analyse a sample you need to use its physical, chemical and biochemical properties to distinguish it not just from the other possible analytes, but also from other substances in the matrix of the sample.  This sounds intimidating, but by breaking it down into separate steps, it becomes a sort of logic puzzle.

   Readily made observations is where this analysis starts. It is the observation of easily ascertainable physical properties, such as the colour, shape and phase of the substance. These can all easily be seen with the naked eye or the aid of a light microscope. These aspects can be surprisingly informative, however care must be taken as powders and pills can easily be mislabelled or tampered with. Some drugs are easy to identify by just their morphology, for example magic mushrooms and marijuana, however others require more attention to detail.

   For example, say you find a brown powder in an unlabelled plastic bag. Based on it's appearance you could identify it as being heroin. However to decide it is heroin would be unwise as there are many other substances it could be. Furthermore, inspecting the substance with a light microscope, rather than jumping to conclusions may reveal the powder to be made of multiple morphologically distinct constituents. Some common drug appearances are as follows:

     Cocaine             White Crystalline Powder
     Ecstasy              Capsules or Tablets of various sizes, shapes and colours
     Heroin               White or Brown Powder often mixed with cutting agents
     LSD                   Very small brightly coloured tablets or pieces of blotting paper

   Once you have noted the physical characteristics, you can proceed to presumptive tests. Most presumptive tests are colour tests, where a small amount of the substance is treated with reagents which will produce specific colours on reacting with the suspected analyte. Typically these tests will produce a positive test in the presence of 1mg of the analyte. A blank test and positive control should always be carried out in order to increase your certainty. A blank test is where you test using the reagents only, while the positive test involves using a pure sample of the analyte along with the reagents.

   Thin-layer Chromatography is one of the most useful presumptive tests. It is a separative technique that provides numerical data about the chemical species in both bulk and trace samples. The process involves dissolving a solid in methanol, spotting it onto a silica gel TLC plate with both positive and negative control samples. The plate is then developed with a 25/6/0.4 by volume mix of methanol, propanone and ammonia. A number of techniques can be employed to visualise the spots, for example spraying the plate with dilute (0.5 M ) NaOH, leaving t dry and then spraying with aqueous Fast Black K.  If this is done, amphetamine spots will appear purple, will methamphetamine will appear orange-red. This is especially useful as amphetamine and methamphetamine appear the same colour in most other presumptive tests.

Thin-layer Chromatography

Thanks for reading, over and out!

Criminal Psychology: What are the Limits of Psychological Profiling?

   Sorry I've been gone for a while but I'm back with an excursion into the world of criminal psychology, much beloved by crime writes (Poirot for example, was basically a very good criminal psychologist). The idea of psychological profiling has always intrigued me, being able to massively reduce the suspects for a crime by deducing the personality and habits of the perpetrator would be an invaluable tool in forensics.

   The first thing to remember however is that a psychological profile can never be used as proof, only as a technique to gain hard evidence. This is because psychological profiling, even by the standards of forensic science, is a very young field with very few standardised methodologies, and even less scientific reliability. this makes the use of a profile as evidence usually a violation of rule 702 of the Federal Rules of Evidence.  Rule 702 states that expert testimony can only be allowed in court when:
 1) the testimony is based on sufficient facts or data
 2) the testimony is the product of reliable principles and methods
 3) the witness has applied the principles and methods reliably to the facts of the case.
   As such there are very few cases where you'd be able to convict on the basis of a criminal profile no matter how well you suspect fits it.

   Even with this caveat, criminal profiling is far from perfect. In fact for a scientific field it's positively riddled with problems. The main problem comes from the fact that the only standardised method of psychological profiling was created by Ressler and Douglas, who were commissioned by the FBI. They interviewed 36 serial killers who had been caught and imprisoned to draw up a list of statistics for the 'average' serial killer. There are several problems with this technique:
   1) The data pool is absurdly small, which leads to large confidence intervals and an inaccurate data pool to draw conclusions from.
   2) The tested group leaves out n important demographic, serial killers who haven't been caught. The 'type' of serial killer who manages to avoid capture (the one where profiling would probably be more needed) is totally absent from the data.
   These inaccuracies can lead to serious mistakes on behalf of the criminal profiler, which could be potentially lead to false convictions or releases if they were taken as hard evidence. For example, there was a case in 2002,of the Washington D.C. snipers, where the killer was predicted to be a white male working alone and turned out to be a father-son duo of black men. "Woops" would be an understatement.

   However there are upsides and success stories. First thing's first, Ressler and Douglas's experiment is ongoing, as the FBI interview more and more serial killers, expanding the pool and aiding with half the problems mentioned above. Psychological profiling can also be used to discover serial killers by linking cases, and showing there is a common killer, and there must be a reason that the FBI continues to fund it.

   Overall Criminal Psychology, while being a useful tool, should never be viewed as hard evidence and regardless of validity will almost always bias the jury. Despite all the criticism listed I believe science gives us the final word in the form of a study by Kocsis which showed that, in detecting criminals, criminal psychologists did have an almost 20% higher validity rate than the closest group, students (72 to 59%). Always consider psychological 'evidence' and always take it with a grain of salt. As always thanks for reading, over and out.

Strychnine

Strychnine is practically synonymous with poison in peoples' minds, due in no small part to crim and murder mystery author. Despite this it is an unlikely choice for a poison as it has an extremely bitter taste.

Name: Strychnine

Chemical Formula: C21H22N2O2

Effect On Victim: Acute Strychnine poisoning effects usually appear very rapidly (10-15 minutes). It starts with muscle stiffness of the back or neck. This is followed by tremors and twitching, and then convulsions. These convulsions are extremely painful, lasting about a minute, and can be accompanied by momentary asphyxiation due to convulsions of respiratory muscles. There's normally a 15 minute rest interval between each convulsion, however the victim will be both exhausted and terrified as a side effect of strychnine include heightened awareness (and the convulsions are violent enough to cause your head to touch your heels, so that's already pretty scary). Death is caused by spasming of respiratory lungs causing asphyxiation.

Strychnine is a scary neurotoxin which mainly affects the nerves in the spinal chord. It binds to neuroreceptors in the spinal chords (it's an antagonist). this causes the neurones to trigger too much as it requires much less glycine, which is the usual trigger, to cause them to activate. This results in the massive convulsions. The victim may also vomit (such a common symptom of poisoning) as that is the body's only way to rapidly excrete the poison.

Lethal Dose:  As little as 30mg can be fatal.

Diagnosis: There are usually no signs of Strychnine in the body post mortem, except that the jaw is occasionally twisted in a death grin. However Strychnine does stay in the body for a long time and can be detected through the use of a dry chemistry process using tandem ion tap mass spectometry. You will find peaks at 334, 319, 306, 277, 261, 246, 233 and 220.

The velos pro dual ion-trap spectroscope is the weapon of choice for identifying strychnine

That's the theory and method behind identifying one of the scariest poison out there. I can see why writers like Agatha christie are so fond of it, it's pretty dramatic. Over and out, as always feel free to comment, until next time!