clinchem blog notes 30 b LAB LETTERS

Most of the time when we hear steroids we think of those drugs which boosts the muscles and turns us into Incredible Hulk’s but doesn’t make our skin green. Our body too is cable of producing its own steroids functioning as hormones and capable of biochemical effects which affects physiologic functions. That is why It very important to develop ways in measuring them to provide substantial diagnosis of disease and treatment.

Thanks to the different leaps towards the advances of science, different analytical procedures are now available.

The most common technique is the COLORIMETRIC METHOD. This method measures steroid metabolites that are present in the urine of the patient.

URINE 17-KETOSTEROIDS

These compounds contain a carbonyl oxygen group at the number 17 carbon on the D ring of the steroid. They are essential compounds for the formation of the different types of steroids thus being labeled “precursors.” Moreover, this analyte is used to evaluate the androgen production.

The specific method used is the ZIMMERMANN REACTION and here’s the simplified procedure.

1. First, 24-hr urine is collected.

2. Then it is hydrolyzed with a strong acid (a mixture of HCl and glacial acetic acid). This process will remove glucoronic acid and sulfate conjugates and it will render the steroid soluble to organic compounds.

3. Extract with dichloroethane.

4. Wash with sodium hydroxide to remove interfering substances

5. Zimmermann reaction takes place when steroid reacts with m-dinitrobenzene in alcoholic KOH.

6. It would look something like this….

Careful though… many drugs may interfere with this test:

URINE 17-HYDROXYCORTICOSTEROIDS

The first method of measuring these substances is the PORTER-SILBER REACTION. This method measures only cortisol, cortisone, 11-deoxycortisol and some of their reduced derivatives. It is also termed as the 17-hydroxycortecosteroid Test. The dihydroxyacetone side chain of the steroid reacts with 2,4-dinitrophenylhydrazine to form a yellow derivative. It is measured spectrophotometrically at a maximum absorbance of 410nm. It also involves the initial steps of preparations like:

1. Hydrolysis with enzymes

2. Extraction to an organic solvent

3. Washing of sodium hydroxide

4. Addition of reagent and heating to allow reaction to take place

To simplify it, here’s the formula:

Again the Zimmermann Reaction may be used for this test. This time, it does not only measures cortisol derivatives but also other cortisol metabolites under the group called 17-ketogenic steroids. However it requires more preparation when it comes to measuring these substances using the Zimmermann reaction.

1. First, glucose is measured using a dipstick. High concentration of glucose may interefere with the results.

2. Specimen must be extracted with ethanol-ether solvent if glucose is higher than 500mg/dl.

3. If not, the assay may be continued.

4. Sodium Borohydride (NaBH4) to convert the steroids to the 17-ketoderivatives.

5. Then add sodium bismuthate (NaBiO3)to remove all side chains

6. Destroy excess sodium bismuthate by mixing sodium bisulfite

7. Hydrolyzed with acid to remove any conjugates

8. Extraction with organic solvent

9. Washing with Sodium hydroxide

10. Solvent evaporation

11. And finally Zimmermann reaction may be performed

Quite a complicated principle for a very simple test right? Aside from the colorimetric procedures other methods are taken advantaged by Clinical Chemistry to measure steroids and they are:

a. Immunoassay Techniques

b. Gas Chromatography

c. High Performance liquid Chromatography

REFERENCE:
Clinical Chemistry: A Fundamental Textbook by Donald Calbreath

Certainly, mandatory drug testing for school admission has its benefits for the society. It attends to the needs of those youth who have drug problems or even prevent the youth from doing so. Sure this is a good thing but it has also downsides.

FIRST POINT. Who’s going to pay for all these expenses? Is it the government or the students themselves? All of us know that in the Philippines there are many families who struggle financially. What happens if they cannot afford to pay for the test? Will the students not be able to attend school? So what happens next? Increased out of school youth? And where would they go if they weren’t in school? Without a doubt, this will be unfair for many students especially for those non-users but cannot pay.

Another question that should be addressed is that what would happen to the students if they turned out positive to the test? Does this mean education is over for them? The government should be able to help the victims of drug abuse in rehabilitating them and ensuring a promising future for them. If not, then the mandatory test would be a total waste time, money and effort.

SECOND POINT. We all know that drug abuse is widespread to the society. So why then only the youth is given mandatory testing? Are we then discriminating them? The reason probably is to detect the problem earlier and prevent chronic use but this should be explained thoroughly. What about the adults? Aren’t they also predisposed to using drugs? Should there be mandatory drug testing for job application then? I’ve learned that it is stated in the law that companies cannot require drug testing for accepting job applicants. If drug testing will be required in school shouldn’t the law stating that companies are not allowed to require employees to be tested be abolished? Then again, this causes public stress.

THIRD POINT. How strict will the drug testing be done? We all know drug testing can be cheated or better yet what if bribery is involved so as to ask officials give negative results for the users? Or what if the users would abstain from using drugs so as to yield negative results temporarily and would use drugs again after the test? Some say use other more sensitive methods. This is pricy for sure. This solution draws us back to problem #1. How can we say that this mass testing would be efficient? If these factors cannot be addressed then the whole purpose of testing is defeated –again a total waste time, money and effort.

FOURTH POINT. What is the coverage of the drug testing? Does it also cover also alcohol and cigarette abuse? From what I know of these two substances are most abused. It also has ill effects for the users. Shouldn’t this problem be addressed also?

THE CONCLUSION. I may say no to mandatory drug testing but it does not mean I encourage the youth to use drugs. Indeed, the proposal has a well defined intention but as I see it, it has many downsides too. Before implementing this rule, it must be discussed and studied properly.

Others say that this proposal would help students in building a good education. I say, there should be other priorities. I am aware that the quality of education is starting to get low. Rather than spending money for testing, why not use it to build classrooms, buy books and scholarships. Isn’t that a more efficient approach in ensuring a bright future?

If we really want a drug free society, it should start from strict government implementation and public observance. The parents also have a major role in preventing their children to become victims. We cannot guard the children 24/7 but proper parenting sets the youth’s positive principles and outlook in life.

“Alcohol and tobacco are the two most abused substances. Each year, in the United States, more than 100,000 people die as a result of alcohol use and over 430,000 as a result of tobacco-related use.”



There was once a time when I said I wouldn’t try any alcohol or cigarette. Well that promise proved me wrong. Thanks to my “good influence” friends, I got my first kick of alcohol. I didn’t like it at first. It didn’t taste any good. I even have to cover my nose so just not to smell its awful spirit. But I observed while drinking, everything just seems so fun –the experience, the laughter and the bonding. And guess what? I didn’t even notice that I was able to finish it and something tells me that I want more.

After we finished, everyone was out of their minds (not drunk) aside from me. I can still stand and walk straight and still have a good perception. This is when I realized that I might have a high tolerance of alcohol. People get surprised when they see me drink. They say it doesn’t fit my personality. I may not look like a drinker, but believe me; this “good boy” can “withstand high doses of alcohol.” I just tell them I am swimming in alcohol when I was still in the womb. Well I guess the saying “don’t judge a book by its cover” is indeed true. But one thing is for sure. I DON’T ABUSE ALCOHOL.

I made another manifesto: I WILL NEVER SMOKE A CIGARETTE. I’m glad I kept this promise of mine. I have friends who smoke. I ask them why they do it and what do they feel. I got a very simple and straightforward answer. “Inalok ako eh. Masarap ang pakiramdam. Nakakatangal ng stress.”

This is probably one of the reasons why most events never run out of alcohol and cigarettes. It just makes the atmosphere and the crowd more invigorating and of course peer pressure. i guess all teens should remember that on the highway of life, whenever conformity hangs a left, good taste is sure to take a mean right.

But of course the major factor why alcohol and cigarettes are heavily abused is because it is
LEGAL. You can buy as much as you want, anytime and anywhere because its very cheap and you can find it anywhere. "Sa dalawang piso, makakaraos kana." Anyone can buy it. I remember before that I was asked to buy cigarette and alcohol from the store and it was easy as 1-2-3. Just hand over the money and there you have it. Take note I was 6 years old at that time. Isn’t there suppose to be an age limit? I know minors aren’t allowed to drink or smoke or even just buy one. But I guess there are many people who are not aware of this. So the third factor would be: rules and regulations are not properly implemented.

You know what is ironic? THIS……