03 September 2016

Lecture #15: TOXIC GASES AND VOLATILES



******  CARBON MONOXIDE  ******

Characteristics

1.      Tasteless, odorless and colorless gas that is produced by incomplete combustion of organic matter.

e.g. automobile exhaust, improperly vented gas heating system, fire

2.      Has 210 times greater affinity to hemoglobin molecule than oxygen. Upon exposure to CO, oxyhemoglobin is converted to carboxyhemoglobin, reducing the delivery of oxygen to the tissues and resulting in tissue hypoxia and anoxia.

Carbon monoxide toxicity: % (v/v) in air

0.01%  – allowable for an exposure of several hours
0.04 – 0.05% – can be inhaled for 1 hour without appreciable effect
0.06 – 0.07% – causing a just noticeable effect after 1 hour exposure
0.1 – 0.12% – causing unpleasant but not dangerous symptoms after 1 hour
                                    exposure.
0.4 and above – fatal in exposure of less than 1 hour

Effects of carboxyhemoglobin

            % Carboxyhemoglobin                              Effect

                        10                                            shortness of breath on vigorous exertion
                        20                                            shortness of breath on moderate exertion,
                                                                        slight headache
                        30                                            decided headache, irritation, fatigue
                        40 – 50                                    headache, confusion, collapse & fainting on                                                                   exertion
                        60 – 70                                    unconsciousness, respiratory failure and
                                                                        death if exposure is continued for a long                                                                        time   
                        80                                            rapidly fatal
                        Over 80                                  immediately fatal


Qualitative determination of carbon monoxide

1.      Alkaline hematin

Dilute the blood 20–40 times with distilled water
Add 0.5 ml of NaOH solution and mix

Result:

Normal blood = (+) greenish brown color formed at once
Blood with CO = retains a pink hue for some time

2.      Katayama’s test

To 2 ml of diluted blood, add 2 ml of yellow ammonium sulfide
Add 2 ml of 30% acetic acid

Result:

Normal blood = (+) green
Blood with CO = remains red

3.      Hoppe–Seyler’s Test

Caustic soda of specific gravity 1.3 produces a greenish color if added to normal blood but retains the bright red color if carbon monoxide is present in the blood.

4.      Kunkel test

The blood, diluted with 4 volumes of water, is mixed with 3 times its volume of 1% tannic acid solution and shaken well. Carbon monoxide blood forms a crimson–red coagulum, which retains its color for several months. Normal blood forms a coagulum which is at first red, becomes brown in the course of one or two hours and then becomes grey in 24 to 48 hours. The blood saturated even with 10% carbon monoxide responds to this test.

5.      Potassium ferrocyanide test

If 15cc of blood is mixed with an equal amount of 20% potassium ferrocyanide solution and 2cc of dilute acetic acid and shaken gently, a bright red coagulum will form, if the blood contains carbon monoxide, while dark brown coagulum will form if the blood is normal.

6.      Palladium test

Carbon monoxide will reduce palladium in solution. In one method based on that reaction, CO gas diffuses into a solution of PdCl2, reducing it to metallic palladium. An excess of KI is then used to convert the unreacted PdCl2 to a solution of colored PdI2, which is measured colorimetrically at 490 mu. The procedure takes several hours, but it is possible to determine amounts of CO as small as 0.1 ul.

A second method, originated by Polis, et.al., involves reacting CO with PdCl2 phosphomolybdic acid in acetone and measuring the molybdenum blue produced.

7.      Winkler solution

The classical method for the analysis of CO and other fluids using the Van Slyke apparatus and Roughton–Scholander syringes should not be overlooked.

Winkler’s solution, containing cuprous chloride, ammonium chloride and copper is used to absorb CO from a gas phase and the change in volume is measured in a microcapillary glass syringe. It is possible to measure as little as 0.02 ml of CO / 100 ml in a 0.5 ml sample, corresponding to the measurement of 0.2 ul of CO. It is also possible to absorb the CO from a gas sample into a solution containing Hb and then to measure the COHb.


Quantitative determination of carbon monoxide

1.      Base on release of gas from hemoglobin complex with subsequent direct or indirect measurement of gas

a.      Gasometric technique
b.      Gas chromatography
c.       Infrared spectrophotometry
d.     Microdiffusion

Carbon monoxide gas is released from hemoglobin by the action of dilute sulfuric acid or lactic–ferricyanide solution. Carbon monoxide then reduces palladium chloride to metallic palladium presenting silver black film on the surface of the reagent.

PdCl2 + CO + H2O ------------> Pd + CO2 + 2HCl

The amount of carbon dioxide is then estimated indirectly by determining the amount of palladium reduced (colorimetric method) or by amount of hydrochloric acid produced by the reaction (titrimetric).

Sulfuric acid was originally used but there are some substances like formic acid that may be converted to carbon monoxide by this method, giving rise to false positive results.

To avoid this reaction, lactic acid, ferricyanide solution has been recommended as the liberating agent.

2.      Estimation of carboxyhemoglobin by it’s typical color absorption bands

a.      Spectrophotometric analysis

(1)   Tietz and Fiereck

A dilute hemolysate of blood is treated with sodium dithionite which reduces methemoglobin and oxyhemoglobin but does not affect carboxyhemoglobin. The absorbance of this solution is measured at 541 nm and 555 nm, the absorbance ratio of A541 nm / A555 nm is calculated and the percent carboxyhemoglobin is determined from the calibration chart.

(2)   IL482 co–oximeter


******  ETHANOL (ETHYL ALCOHOL)  ******

Characteristics

1.      In gastric and intestinal mucosa, it is readily absorbed 1 hour after ingestion.

2.      In lungs and kidneys, it is excreted unchanged.

3.      In the liver, it is primarily metabolized

4.      In the nervous system, ethanol’s toxic form, acetaldehyde is known to give its toxic effect.

Toxicity ratings in ethanol poisoning

1.      35 mg/dl – will interfere with judgement of distance, speed and causes euphoria.

2.      50 – 100 mg/dl – intoxicated to operate a motor, characterized by euphoria and loss of normal social inhibitions.

3.      200 – 300 mg/dl – correlated with mood exaggeration, slurred speech, apathy, disorientation and loss of coordination

4.      400 – 500 mg/dl – coma and death


Symptoms and actions in acute alcohol poisoning

1.      0.2% or less

a.      Release of inhibition leading to talkativeness, boisterous behavior, delayed reflexes, minor muscular incoordination and sometimes nystagmus
b.      Marked cutaneous dilatation
c.       As the blood concentration is increased, muscular control and mental activity becomes increasing difficult


2.      0.25 – 0.3%

a.      Blurring of vision
b.      Lack of acuity of the conjunctiva
c.       Reddening of the conjunctiva
d.     Dilatation of the pupils
e.      Diplopia (double vision)
f.        Difficulty of focusing
g.      Nystagmus (involuntary surging movement)


3.      0.4 – 0.5%

a.      Complete loss of motor activity
b.      Stupor or coma may intervene
c.       Shallow and slow respiration
d.     Death may occur from respiratory failure
e.      Circulation remains relatively intact except for the effect of cyanosis


Adverse reaction of ethanol ingestion

Respiratory disease like pneumonia is common among alcoholic due to increase heat loss and respiratory depression.

Habitually heavy drinker shows considerable tolerance so that higher concentrations are required for impairment of function.

On recovery from alcohol intoxication, acidosis, edema of the brain and swelling of the liver and kidneys have been observed. Headache noted after is due to edema of the brain. Liver function is diminished during alcohol intoxication and the liver becomes susceptible to injury by chemical agents like carbon tetrachloride or chloroform.

Death due to alcohol depression is due to respiratory failure and occurs 1–10 hours after taking about 24 hours or more for one to recover from alcohol depression.


Fate and excretion

Ethyl alcohol is rapidly distributed to all the tissues in nearly equal concentrations. At equilibrium, the brain contains slightly more than the blood, while the urine contains 1.2 to 1.3 times as much as the blood.

Less than 10% of ingested alcohol finds its way into urine and expired air. The remainder is burned in the body to carbon dioxide and water at a rate of about 8 grams per hour. Blood alcohol and probably tissue alcohol decreases at the rate of from 170 – 270 mg/kg/hr. The intermediate product in the oxidation of alcohol is acetaldehyde.


Method of determination

1.      Gas–liquid chromatography

2.      Enzymatic analysis using alcohol dehydrogenase

Ethanol is oxidized in the presence of ADH to acetaldehyde. In the course of the reaction, NAD, a coenzyme is reduced.

C2H5OH + NAD ----------> CH3CHO + NADH + H+

The increase NADH can be measured by the increase in absorbance at its absorption maximum at 340 nm.

Equilibrium for this reaction lies strongly to the left. At neutral pH and at normal NAD concentration less than 1% of ethanol present is oxidized to acetaldehyde.

The reaction however can be driven almost completely to the right by maintaining a high pH and removing the acetaldehyde as it formed by reacting it with semicarbazide.


Precaution in specimen collection

1.      Serum, urine or saliva can be used as a specimen
2.      Sodium fluoride is the anticoagulant of choice if plasma will be used.
3.      If blood will be used, the venipuncture site should be cleaned with a disinfectant that does not contain alcohol
4.      Specimen should be analyzed immediately or kept stoppered and refrigerated until tested.
5.      Serum, plasma and saliva concentrations of ethanol are 20% higher than those of whole blood.


******  METHANOL (METHYL ALCOHOL)  *****

Characteristics

1.      It is used as solvents in paints, varnishes and paint removers.

2.      It is also used as an anti–freeze fluid.

3.      Poisoning is usually due to accidental ingestion by children or by alcoholism.


Symptoms of toxicity

1.      Nausea and headache or progressing to convulsion and coma and blurring of vision, which may progress to temporary or permanent blindness.

2.      Death may occur from ingestion of as little as 10 to 30 ml.

3.      Methanol is converted to formaldehyde and formic acid in man, resulting in an accumulation of formic acids and other acids. This reduces the alkali reserve resulting in a metabolic acidosis.

4.      Elevation of amylase due to necrosis in the pancreas may also be noted.


Methods of determination

1.      Gas–liquid chromatography

2.      Colorimetric method using CTA

Methanol is oxidized to formaldehyde with permanganate. Chromotropic acid, CTA (4,5–dihydroxynapthalene–2,7–disulfonic acid), which is specific for formaldehyde, is then added for development of purple color.

Disadvantages of drawbacks to the method

a.      Methanol is not quantitatively oxidized to formaldehyde. Further reaction may also take place in which formaldehyde can be oxidized to formic acid and further to carbon dioxide.

b.      The presence of reducing substance other than methanol will affect the system so that the procedure can no longer be applied quantitatively. Alcohol is the most common substance that interfere in cases of methanol poisoning

The above drawback can be obviated by

(1)   The procedure must be carried out in the same manner for both standards and unknown.

(2)   An excess of ethanol is added to both standard and unknown resulting in a constant interference.

c.       A minor drawback is the use of concentrated sulfuric acid for the development of the final color.

A false high result may be obtained due to the dehydrating effect of concentrated sulfuric acid on appropriate organic compounds. This is encountered occasionally in patients with severe acidosis. Some substances may appear in TCA filtrate such as glycolic acid which reacts as follows:

H2C(OH)COOH ------------------> HCHO + CO + H2O

The above interference can be detected by running a blank for comparison. This is done by running the same procedure to a portion of the filtrate but the oxidation procedure is omitted. Any color that develops in the unoxidized specimen is due to formaldehyde or glycolic acid.

Methanol can be separated by subjecting the specimen to steam distillation or microdiffusion in a Conway unit

3.      Other methods

a.      Hindbery and Wierth method
b.      William et.al method


*****  BARBITURATES  *****

Malonyl carbamide (barbituric acid) was first synthesized by Adolf von Baeyer in 1863.

In 1903, Emil Fisher and Joseph von Mering synthesized diethyl barbituric acid (barbitone) from urea and malonic acid. This was the first barbituric acid compound with hypnotic qualities and marketed under the trade name Veronal. While over 1,500 different barbiturates have been prepared, only a few are used in medicine today.

Medicinal value:

1.      Prescribed for insomnia and epilepsy

2.      Prescribed for treatment of high blood pressure

3.      For diagnosis and treatment of mental illness

4.      Used for releasing patient before and during surgery

5.      To relieve pain


Classification of barbiturates

1.      Long acting barbiturates – are distributed more evenly and have long elimination half–lives, making them useful for treatment of epilepsy.

e.g. Phenobarbital, diallybarbituric acid

2.      Intermediate acting barbiturates

3.      Short acting barbital

4.      Ultra–short acting barbiturates – are highly lipid–soluble and rapidly penetrate the brain to induce anesthesia, then are quickly redistributed to other tissues. For this reason, the duration of effects is much shorter than the elimination half–life for these compounds.

e.g. hexobarbital, thiamytal, thiopental


Mechanism of toxicity

All barbiturates cause generalized depression of neuronal activity in the brain. These effects are primarily mediated through enhanced GABA–mediated synaptic inhibition. Hypotension that occurs with large doses is caused by depression of central sympathetic tone as well as by direct depression of cardiac contractility.


Symptoms of toxicity

1.      Obstruction of the airway by vomitus, secretions, laryngeal spasms and the relaxed tongue.

2.      Head injuries, internal injuries, burns, scalds, and fractures may result from falling.

3.      Bronchopneumonia, pneumonitis, pulmonary edema and atelectasis may develop.

4.      Acute heart failure and failure of peripheral circulation (circulatory collapse) may occur.

5.      Vomiting and lack of fluid intake can cause serious disturbance of electrolyte and water balance.


Toxic dose

Small amounts of barbiturates taken in produces relaxed sociable feeling and decrease alertness and slower reactions.

Larger amounts of barbiturates consumed will produce sluggishness in person. He becomes gloomy and often quarrelsome. Muscular steadiness, tremor, hypotenia, difficulty of speech and sometimes excitement are noted. There may be flushing of the skin and he will gradually slump into a deep sleep.

Much larger doses may result in coma and may exhibit hyperthermia or pinpoint pupils. At first, the unconsciousness is associated with slow and shallow breathing, cyanosis, normal or dilapidated pupil which react to light if depression is not too deep; nystagmus, a diminishing blood pressure and loss temperature. The EEG waves are slow in deep sleep due to barbiturates but in coma, the voltage is low with rapid rhythm superimposed on slow waves.

The vasomotor and respiratory centers are depressed to a greater degree than with alcohol. Pre–medication doses of amobarbital or pentobarbital in aged person with hypotension maybe associated with a fall of 20 – 100 mm/Hg.

Smooth muscles are inhibited and visceral ganglia are depressed.

Urinary excretion of reducing corticoid is doubled but there is no change in 17–KS excretion in barbiturate poisoning.

Barbiturates and analgesics are said to be dangerous in the presence of porphyria and should not be used to relieve pain associated with the disease.

The danger is said to be dangerous in the presence of porphyria and should not be used to relieve pain associated with the disease.

The danger is said to be associated with the increased secretion of anti–diuretic hormones produced by such mixtures. Barbiturates are especially dangerous when taken along with alcohol because the depressant action of one multiples, the depressant action of the other leading to death.

Intravenous administration of the more concentrated solution occasionally causes venous thrombosis if inadvertently the needle punctures through the skin or fails to enter the vein.

A phenomenon occasionally associated with coma due to barbiturates and other depressants is dermatitis characterized by patches of erythema surmounted by bulbous vesicles.
  
Fate and excretion

Barbital is excreted into the urine mostly unchanged, 75% being excreted in 48 hours. It can be identified in the urine as early as 1 hour after ingestion.

Only 25% of a large of phenobarbital case be recovered from the urine; the remainder is destroyed in the body.

The liver seems to be the chief organ of detoxification. Hyperthyroidism hastens the process.

Cause of death

From oral dose         =          acute early death is due to respiratory illnesses
Delayed death          =          usually due to pneumonia

Intravenous administration may cause death from circulatory collapse.

The severity of depressant action of barbiturates will depend upon the following factors:

1.      Dose of barbiturates
2.      Mode of administration
3.      Degree of tolerance
4.      Presence or absence of other drugs in the body
5.      State of excitability


******  OPIUM, OPIUM DERIVATIVES AND NEWER SYNTHETIC ALKALOIDS  *****


Opium is the dried juice obtained from the unripe seed capsule of the opium poppy, Papaver somniferum as well as the stem.

Morphine makes up 10% of the weight of the official opium (from Asia minor) while about 1–2% is made up of codeine, noscapeine, papavereine, thebaine and a number of minor alkaloids.

Drugs artificially prepared from morphine

1.      Diacetylmorphine
2.      Dihydromorphine

Synthetic substrate for morphine and codeine

1.      Meperidine (Demerol)
2.      Methadone
3.      Dihydrocodeinone
4.      Dihydrohydrocodeinone
5.      Levorphan
6.      Anileridine
7.      Pentazocaine
8.      Propoxyphene

Absorption

            Opium alkaloids are moderately absorbed from the alimentary tract
            Morphine is 2–4 times as toxic when given hypodermically.
            Concentration of 1 mg / ml of blood represents a large dose.

Toxic dose

1.      Fatal dose – 2–5 grams or morphine and 0.07–0.5 grams of its salt represent dose in robust adults. One milligram of morphine has been fatal to an infant. Ten milligram of morpine given hypodermically to and aged has caused respiratory failure.

2.      Codeine – 0.5 mg is often fatal

3.      Diacetylmorphine (heroin) – 2–3 times as toxic as morphine

4.      Dihydromorphine (Dilaudid) – 5–10 times as toxic as morphine.


Symptoms of toxicity from morphine

1.      Slow respiration, shallow or sighing or Cheyne–Strokes type

2.      Unconsciousness

3.      Cyanosis

4.      Construction of pupil

5.      Hyperglycemia and glycosuria may be present

6.      Perspiration is often present

7.      When depression become deeper and the respiration slower and inadequate, prostration with loss of reflexes and relaxation of the muscles takes place, skin becomes pale, cold and moist; pupils dilate; heart beat becomes slower and latter irregular, rapid and weak; convulsion may be observed due to asphyxia and morphine stimulation of the cord.

8.      Death is due to central respiratory paralysis

9.      If the patient recovers, he experiences physical and mental fatigues for several days. He also suffers from obstinate constipation.

10.  Occasionally, a person may respond atypically to a therapeutic dose. Instead of gradual development of cerebral depression, there is cerebral excitement, nausea, vomiting, headache and even delirium.


A.    Codeine

1.      Minimal lethal dose of codeine is reported to be 80 mg.
2.      In general, codeine and morphine behaves similarly in the body.
3.      Cerebral and cord hyperirritability are more marked with codeine compared to morphine so that delirium and convulsion may occur during coma.
4.      Pupils are not contracted as with morphine and are dilated during delirium.
5.      Constipation is less compared to morphine.
6.      Nausea is more frequent compared to morphine
7.      Respiratory and circulatory effects are practically identical.


B.     Diacetylmorphine

1.      Large proportion of acute poisoning occurs among addicts who have taken a dose beyond their tolerance.

2.      Fatal dose:           10 – 600 mg%


C.    Synthetic substitutes

1.      Dihydroxyhydroxycodeinone – hallucination effects

2.      Dihydroxycodeinone – often causes dizziness

3.      Meperidine – causes mydriasis and release histamine from mast cells.

4.      Pentazocine – dose of 30 – 60 mg by injection causes drowsiness, dizziness, euphoria, rhinorrhea, incoordination, weakness, tremors, convulsion, allergic edema, insomnia and hallucination.

Etiology

a.      Addiction developed through the medicinal use of drug
b.      Influence of the drug through vice.

Symptoms and actions

a.      Stimulation and depression of the CNS simultaneously.

b.      Tolerance development occurs more readily toward the depressant effect than toward the excitant effect.

c.       Repeated administration of morphine leads to less euphoria and depression of the dose remains constant.

d.     Excessive dosing is usual; there is mental dullness, contraction of pupils and certain hypersensitiveness to sudden stimuli.

e.      Skin is usually dry.

f.        Diminish salivary excretion.

g.      In women, menses are diminished or suspended.

h.      Conception is rare.

i.        Decreased libido.

Sudden withdrawal of the drug:

a.      Feeling of anxiety, restlessness, furtiveness, dilation of the pupils.

b.      Followed by vague or sometimes acute pain the abdomen, skeletal muscles and joints.

Chronic poisoning by opium derivatives

Chronic use leads to a physical dependence, a condition usually called addiction. Instead of using addiction and habitation, the WHO used the word drug abuse and drug dependence.

Drug abuse – persistent or sporadic use of excessive drug which is inconsistent with unrelated to acceptable medical practice.

Drug dependence – a state, psychic or sometimes physical, resulting from the interaction between a living organism and a drug, characterized by behavioral and other responses that always include a compulsion to take the drug on a continuous or periodic basis in order to experience the psychic effects and sometimes to avoid the discomfort of its absence.


Difference between CODEINE and MORPHINE

                                          CODEINE                             MORPHINE

Excess alkali hydroxide                  precipitated                          soluble
Excess alkali ammonia                    soluble                                   precipitated
Iodic acid                                           does not reduce acid           reduces acid
Neutral FeCl3                                    no color produced               blue color
Ferric chloride                                  Prussian blue                                    Prussian blue
Potassium ferricyanide                   negative                                 positive


Test for Morphine

1.      Preliminary Test

a.      Acidic portion of the aqueous alkaline solution with sulfuric acid.

b.      Add a few drop of iodic solution

c.       Extract with chloroform

d.     Result:

A violet color at the choloroform layer is positive for morphine

2.      Nitric Acid Test

a.      Dissolve morphine in nitric acid solution

b.      Orange color – positive for morphine

3.      Froehde Test

a.      Dissolve morphine in Froehde’s reagent

b.      Result:

Fine violet color changing to blue to green and finally red.

4.      Ferric chloride test

a.      Add a few drops of FeCl3 to a neutral solution of morphine.

b.      Result: Blue color is positive for morphine

5.      Marquis Test

a.      Dissolve morphine in Marquis reagent

b.      Result: Purple red color changing to violet to pure blue

6.      Selenious sulfuric acid test

a.      To a portion of the dried extract, add a few drop of freshly prepared 0.5% solution of selenious.

b.      Result: Formation of a green color is positive for morphine. The test is said to be positive to a 0.01 mg of morphine.

7.      Microscopic test

a.      Dissolve a portion of the extract in 1–2 gtts of 1/10N HCl.

b.      Place a drop of the solution on a glass slide and a drop of Wagner’s reagent.

c.       The slow formation of red brown plate or large needles one or both sides of which are deeply serrated shows the presence of morphine.

d.     Marme’s solution precipitates clusters of slender needles from solution of morphine as dilute as 1:1000.

e.      The detection of alkaloid of opium can be performed on the residue of the extract with alkaline chloroform and amyl alcohol followed by performance of the color reaction of alkaloids.

f.        For each of these tests, a small portion of the residue remaining after evaporation of the alkaline chloroform extract should be dissolved in a few drops of dilute HCl.

g.      A reaction specific for apomorphine is purple red with Fluekiger’s reagent

(1)   Codeine – green brown color with Mandelin’s reagent
(2)   Morphine – purplish red with Mandelin’s reagent and Mecke’s reagent


(3)   Alkaloids  can also be separated from each other by paper chromatography



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