03 September 2016

Lecture #16: THE HEAVY METALS





******  LEAD  *******

The major source of lead exposure in modern times is the lead–based paint. Lead is used in type metals, storage batteries, paints, toys and in gasoline industry.

Sources of exposure

1.      Industrial workers are exposed to lead fumes and dusts in smelting plants to lead salts in the pigments and storage batteries as well as to lead silicate in potter industry.

2.      Inhalation represents the most serious made to exposure

3.      Children are exposed to lead containing paints.

Symptoms

1.      Loss of appetite, headache, irritability, neurological symptoms and intermittent vomiting.

2.      Finally encephalopathy with cranial nerve paralysis, delirium and coma may occur.

3.      Basophilic stippling of red cells in peripheral blood.

4.      X–ray evidence of dense band at the epiphysis of the long bones of children represents additional evidence of lead poisoning.

5.      More evident sign of plumbism are lead lines in the gums, emaciation, anemia and increase CNS symptoms. Since lead is secreted very slowly, it accumulates in the tissues to toxic concentrations.

6.      Increased urinary excretion of porphyrin. Porphyrinuria is usually earlier found and more frequent than erythrocytic stippling.


Mechanism of toxicity

The toxic effects of lead are due its ability to non–competitively inhibit many enzyme systems. The effects of lead on heme synthesis and the resultant anemia are directly related to methods of clinical laboratory diagnosis. Lead inhibits the activity of several enzymes in the biosynthetic pathway for heme, including delta– aminolevulinic acid (ALA) dehydrase and ferrochelatase. The substrates of these enzymes, ALA and protophorphyrin IX, accumulate in red blood cells. When ferrochelatase is inhibited or when iron is not available, excess protoporphyrin IX combines with zinc in the erythrocyte produces zinc protoporphyrin (ZPP).

Methods of determination:

1.      Tompsett colorimetric analysis
2.      Hematofluorometry
3.      Flameless Atomic Absorption Spectrophotometry
4.      Anodic Stripping Voltammetry


******  THALLIUM  ******

Thallium is being used as rodenticides by professional exterminators. The principal compound responsible for poisoning is thallium sulfate and thallium acetate. The thallium salts are absorbed from the skin as well as from the digestive tract.

Toxic dose:

1.      0.12 – 60 g – for adults
2.      8.5 mg / kg body weight – for children
3.      8 – 10 mg – will cause epilation

Etiology of poisoning

1.      Accidental ingestion of poisoned grained intended for rodent extermination.
2.      Suicidal
3.      Homicidal
4.      Medication or cosmetic to depilate the skin

Symptoms and actions

1.      Acute poisoning from a single dose, nausea and vomiting may occur early, but oftentimes the symptoms occur several hours after.

2.      Metallic taste, nausea and vomiting

3.      Dryness of the mouth, soreness of gums

4.      Rhinorrhea and conjunctivitis

5.      Puffiness of eyes and face

6.      Insomnia, deafness and scrotomas

7.      Tingling pains on the feet and hands as well as muscle sores.

8.      In a few days, severe stomatitis and paralysis of one or more muscles may occur.

9.      Loss of hairs occurs within three weeks, leaving only a fringe of hair along the forehead, the eyebrows and the pubic hair are characteristically retained.

10.  Eight weeks after poisoning, white strips across the nails are seen.

11.  Mental and neurological symptoms occur

12.  A gingival line is sometimes seen

13.  Delirium, convulsion and coma are terminal symptoms.

Fate and excretion

Thallium is a cumulative poison and is found in the liver, kidney, bone, muscle and in highest concentration in epididymis.


Pathology

Inflammation of the gastro–intestinal mucosa and parenchymatous degeneration of the liver and kidneys are seen in acute poisoning. Marked loss of hair and severe anemia may be noted. Eosinophilia has been reported. Permanent organic changes in the brain may occur.


Methods of quantitation

1.      Fluorescence test

Dilute 1 ml of the solution with 9 volume of saturated NaCl in a small beaker. Expose to the ultraviolet light of a mineralized ultraviolet lamp. A bright blue fluorescence is given by a quantity of thallium as small as 1 mg.

2.      Microscopic test

a.      To a drop of solution on a glass slide and a drop of dilute HCl, these forms a precipitate of white irregular crosses and clusters of radiating crystals.
b.      Uranyl acetate yields a precipitate of needles and prisms with albumin.

3.      Action of bromine on Thallium

Thallium is converted to its oxidized form by the action of bromine water. After destruction of excess bromine, the thallic ion is complexed with methyl violet to form a blue to violet compound of unknown structure that is soluble in benzene

Procedure

a.      Into a glass–stoppered tube, place 1 ml of urine and 3 drops of concentrated HCl. Mix.

b.      Add 5 drops of bromine water. Mix thoroughly.

c.       Add 5 drops of 20% sulfosalicylic acid to decolorize the bromine.

d.     Add 1 drop of methyl violet solution and mix.

e.      Add 1 ml of benzene, stopper the tube and shake thoroughly. After separation of the layer, decant or aspirate of the benzene and observe its color if any.

Interpretation

A colorless benzene layer rules out the presence of 0.3 ug or more of thallium

A positive test imparts a blue violet color to the benzene

No interference is seen with levels up to 1 mg of borate, oxalate, chlorate, nitrate, phosphate, sulfate, chlorides, bromide, perchlorate or EDTA.

Color formation is inhibited by 1 mg quantities of nitrite, sulfite, sulfides, thiosulfates and thiocyanate. The only metal that gives a false positive is 0.1 mg of mercury.


******  ARSENIC AND ARSINE  ******

Arsenic is used in ant poison, insecticide, weed killers, paint, wall paper, ceramics and glass. The action of acids on metals in the presence of arsenic forms arsine gas. Alloys such as ferrosilicon may release arsine upon contact with water since the ferrosilicon may be contaminated with arsenic.

The fatal dose of arsenic trioxide is about 120 mg. The allowable food residue is limited by federal law to 1.4 mg /kg. The exposure limit for arsine is 0.05 ppm; for arsenic acid, arsenates, arsenites and other compounds or arsenic is 0.5 mg/m3.

Arsenic presumably causes toxicity by combining with sulfhydryl (–SH) enzymes and interfering with cellular metabolism.


Clinical findings

The principal manifestations of arsine poisoning are hemolysis.

1.      Acute poisoning

a.      Ingestion – violent gastroenteritis, burning esophageal pain, vomiting, bloody diarrhea.

b.      Inhalation may cause pulmonary edema, restlessness, dyspnea, cyanosis, cough with foamy sputum.

c.       Exposure causes burning and stinging of the face, tightness of chest, nausea, dysphagia, hemolysis, hemoglobinuria, bronzing of the skin, enlargement and tenderness of the liver and spleen

2.      Chronic poisoning

a.      CNS – polyneuritis, optic neuritis, anesthesias and parasthesias.

b.      Skin – bronzing, alopecia, localized edema and dermatitis

c.       Gastrointestinal tract – cirrhosis of the liver, nausea, vomiting, abdominal cramps, salivation

d.     General effects – anemia, weight loss, aplastic anemia

e.      Cardiovascular system and kidneys – chronic nephritis, cardiac failure, dependent edema.

f.        Visual impairment and optic atrophy

g.      Arsenic and its compound are carcinogenic


Detection of arsenic

The test commonly used as referred to as the Reinsch test. It depends on the fact that metallic copper in the presence of acid will reduce arsenic to the elemental form. The arsenic deposits on the copper has a visible dark film.

            3Cu     +          2As ------------------> 3Cu2+     + As

The oxidized form of antimony, bismuth, mercury and selenium can also be reduced by metallic copper under this condition.

Rieder’s modification

Reagents:

1.      Concentrated HCl
2.      Copper spiral – wind bright, clean copper wire around 3mm glass rod, about 8 – 10 times to make a tight spiral.

Procedure

1.      Place 100 ml of urine in a shallow dish
2.      Add 10 ml of concentrated HCl.
3.      Boil the solution until the volume is reduced to about 20 ml.
4.      Remove the copper, rinse gently with distilled water, examine and note any color change.

Interpretation:

If copper is still bright red, arsenic, mercury and selenium are ruled out

Arsenic, selenium                gray to black
Mercury                                 light gray to silvery and become shiny on rubbing

Other substances that may produce a gray to black color are: some Sulphur compounds, antimony, bismuth, tellurium

In the case of a positive result, the nature of the deposit on the copper must be verified by further test.

Arsenic can be quantitated after wet digestion of another specimen by colorimetric method.

Recently, the technique of atomic absorption spectroscopy has been used.

Normal arsenic levels in urine are less than 50 ug / ml.

Chronic poisoning – arsenic levels will rise to 100 ug / l

Acute poisoning – 1 mg / l or more

Since arsenic is readily bound by sulfhydryl groups in proteins, considerable arsenic is bound by keratin and subsequently deposited in hairs and nails.

Minimum requirement – 1 gram of clean hair clipped close to scalp should be submitted.


******  TEST FOR HEAVY METAL  ******

Procedure:

1.      10 ml of macerated liver or kidney or urine or gastric contents is placed in a small Erlenmayer Flask. Set up a positive and negative control.

2.      Ad 2 ml of concentrated HCl and small copper sheath (5x10 mm). The copper must be shiny clean to show clearly a contrast of a deposition of a heavy metal if positive. A copper wire spiral may be used instead of the copper sheet.

3.      Cover the flask with a watch glass and heat gently for about 1 hour, avoid boiling to prevent rapid evaporation. If large amount is present, deposits may occur in less than 1 hour. After 1 hour, note the appearance of the deposits.


Mercury                     shiny silver deposits           0.50 mg / 10 ml
Arsenic                       dull back deposits               0.010 mg / 10 ml
Bismuth                     shiny black deposits            0.020 mg / 10 ml
Antimony                  dark purple sheen               0.020 mg / 10 ml


The amount should be reported as negative, small, moderate or large to differentiate the dark deposit from each other.

Mercury – dull black – place a copper in a small amount test tube and add 15 drops of 10% potassium cyanide. The dark deposits due to arsenic will dissolve. If large amount of arsenic deposits is present, more cyanide is required to dissolve it. This test for arsenic is very sensitive. Antimony or bismuth deposits are not dissolved.

Bismuth – shiny black – place the copper and deposit into a small test tube and add about 15 drops of 15% sodium sulfite and 1 ml of 15% nitric acid. The deposits due to bismuth dissolves while that of arsenic or antimony will not. To the dissolved bismuth solution, add 1 ml of water and 1 ml of bismuth test reagent – orange turbidity. Bismuth test reagent consist of 1 gram of quinine sulfate dissolve in 100 ml of 0.5% HNO3; then dissolve in 2 grams potassium iodide into solution. This test is specific and sensitive to 20 ug of bismuth.


Antimony – purple shiny and unchanged by both treatment.





No comments: