Chapter III
NON VOLATILE POISONS
This group of poisons includes the organic and alkaloidal substances which are non-volatile with steam from acid solutions but are extracted by acidulated alcohol.
This method which has been used for the isolation of non-volatile poisons was first devised by Stas for the extraction of nicotine from viscera. It has been modified by Otto and many worker and is now known as the STAS-OTTO Process. The essential facts on which the process is based are:
1. Almost without exception, these substances which are non-volatile with steam, are soluble in alcohol and to a greater or lesser extent in acidified water or hot neutral water.
2. From aqueous solutions these substances may be extracted by repeatedly shaking with an immiscible organic solvent, the solution being acidified for the extraction of neutral or acidified substances and made alkaline for the extraction of basic substances like the alkaloids.
The STAS-OTTO process was later modified by Autenrieth and this procedure has been adopted by the Forensic Chemistry Division of the National Bureau of Investigation. The procedure consists in making the finely ground material acid with tartaric acid and thoroughly mixed with 2-3 times as much 95% alcohol. The materials used for the steam may be utilized for this procedure. The mixture is then heated over a water bath with a reflux condenser for about half hour. The contents of the flask is cooled and filtered to remove fat as completely as possible. The filtrate is then evaporated to a thin syrup in an evaporating dish over a water bath. The residue is mixed with 100mL distilled water and filtered. Addition of water would cause the separation of fats and resinous matter. The filtrate is again evaporated to dryness or thin syrup over a water bath and the residue mixed with 95% alcohol. A whitish substance, more or less viscous or slimy consisting mostly of protein (albumin, albumoses, peptones), dextrin like compounds, and inorganic salts will separate. Tartrates of the alkaloids and other organic poisons dissolved. The larger the volume of alcohol used, the more complete is the precipitation of these substances that interfere with the detection of poisons. Evaporate the filtrate over a water bath and take the residue with about 50mL of distilled water. If the solution is not perfectly clear, filter again through a moistened filter paper.
This aqueous solution is now examined according to the STAS-OTTO process in the following manner.
1. Thouroughly extract the acid solution several times with ether. This solvent will dissolve free acids and acid compounds such as picric acid, salicylic acid, barbiturates; neutral substances such as acetanilide and phenacetin; very weak bases such as antipyrine, caffeine and nicotine.
2. Make the acid solution strongly alkaline with NaOH and again extract first with ether. All alkaloidal poisons except phenolic bases such as morphine and apomorphine will be extracted. Then extract with chloroform to complete the extraction of alkaloids not completely extracted with ether.
3. The aqueous solution alkalinized with NaOH is now changed to one alkaline with ammonia by addition of ammonium chloride. Extract first with ether and then with hot chloroform. Ether will extract apomorphine and traces of morphine. Chloroform will extract morphine, narceine, and other bases not very soluble in ether.
A. EXAMINATIPON OF THE ETHER EXTRACT OF THE AQUEOUS ACID SOLUTION
PICRIC ACID
Synonyms: Trinitrophenol
Lethal dose: 2 grams
Properties: It is bright yellow, odourless crystalline substance with a bitter taste. It has a tendency to explode when heated or subjected to percussion.
Uses: It has the property of dyeing silk and wool. In medicine it is employed as an external application in the treatment of burns and skin diseases and sterilization of the skin before a surgical operation.
Toxic Action: Picric acid is quite a vigorous poison. When taken internally, it produces a striking yellow pigmentation first of the conjunctiva and then of the entire skin, usually designated as “picric acid icterus.”Picric acid and its salts like most nitrous compounds decompose red blood corpuscles for methemoglobin. Consequently compounds of picric acid are blood poisons. It is also a convulsive poison, for it irritates the CNS causing convulsions. Finally it exercises its power of precipitating proteins in acid solution. This manifests itself by necrotic tissue changes especially in those organs of the body that have an acid reaction, such as the stomach and kidney.
Symptoms:
1. Pallor, cyanosis, dizziness, fatigue dyspnea
2. Headache, pain in the legs, sensitization
3. Irritation and corneal damage
4. CNS depression
5. Toxic jaundice, liver enlargement, alter liver shrinkage
6. Fatty degeneration of the kidney
Treatment:
1. Stomach must be washed out with milk and egg whites
2. Intravenous injection of dextrose should be given to aid in the reduction of picric acid.
3. Give analgesic to relieve pain
Detection:
1. Isopurpuric Acid test
Gently heat an aqueous solution of the picric acid (1mL of 5% solution) with 5gtts of aqueous potassium cyanide solution.
2. Picramic Acid Test
a. With dextrose – warm 1mL of the acid solution with 2-3 gtts of sodium hydroxide and 2-3 gtts of glucose solution.
b. With Ammonium Sulfide – To 1mL of picric acid solution, gently add a few gtts of ammonium sulphide and sodium hydroxide solution. Warm the solution gently.
3. Ammoniacal Copper Test
Add 5gtts of ammoniacal copper sulfate solution to 1mL of picric acid solution
4. Dyeing of Thread test
Heat 2mL of the picric acid solution and then place white thread of wool, silk, cotton and synthetic fibers in the solution for a few hours. Remove the threads and then rinse them thoroughly in clean water. Observe the result. Mount threads treated with picric acid.
ASPIRIN
Synonyms: Acetyl salicylic acid, 2-acetoxybenzoic acid, ASA
Toxicity Rating: Moderately Toxic
Properties: This is the aceto-salicylic acid formed by the action of acetic anhydride or acetylchloride upon salicylic acid. It is white, inodorous, crystalline powder having sweet, acidulous taste and with a melting point of 137°C. It is about one and one-half times more toxic as sodium salicylate in man.
Lethal dose: 0.2-0.5g/kg
(35 tablets of 300mg ASA or 130 tablets of 80mg ASA)
Uses: Analgesic, antipyretic, anti-inflammatory, anti-rheumatic, fungistatic, rubefacient.
Toxic Action: Aspirin causes the toxic syndrome “salicylism”. It can also produce poisoning by penetrating intact skin. It is a derivative of salicylic acid. The major toxic signs and symptoms arise from the stimulation and terminal depression of the CNS. Aspirin irritates the mucous membrane of the stomach, irritates the intestines without decomposition. After 30 minutes to 1 hour of ingestion some of it may be found unchanged in the urine. The remainder is hydrolyzed to salicylic acid which is excreted partly unchanged, partly glucuronide conjugates after conversion of salicylic acid to gentisic acids. A toxic effect which may occur even with small doses is irritation of the gastric mucous leading to gastro-duodenal bleeding. In large doses it produces acidosis. An aspirin splitting enzyme is found in the blood plasma, urine and saliva.
Symptoms:
Acute Poisoning
1. Burning pain the throat, stomach and abdomen
2. Slight to moderate hyperpnoea, lethargy, vomiting
3. Tinnitus, hearing loss and dizziness
4. Dimness of vision, fullness in the head and headache.
5. Irritability, restlessness, confusion and delirium
6. Fever, sweating, and dehydration
7. Cyanosis, oliguria, uremia, pulmonary edema
8. Convulsions, coma and respiratory failure.
Chronic Poisoning
1. Tinnitus, abnormal bleeding (gastric or retinal)
2. Gastric ulcer, weight loss, mental deterioration and skin eruptions.
3. Liver damage
4. Acetone-odor breath and urine
Treatment
1. Administer emetic using syrup of Ipecac
2. Delay absorption of the remaining poison by giving activated charcoal.
3. Do gastric lavage with water or 1% Na bicarbonate solution (mild alkali delays salicylic absorption)
4. Use saline cathartic such as sodium or magnesium sulfate.
5. Correct dehydration and hypoglycaemia with infusion of dextrose of water (D5W) or isotonic saline solution (PNSS).
6. Administer an osmotic diuretic such as mannitol (useful to prevent systemic alkalosis) to treat cerebral edema if coma persists even if salicylate poisoning has returned to normal.
7. If blood pressure is low, give 10-15mL/Kg of whole blood by transfusion over a period of 1 hour.
8. Treat respiratory depression by administering artificial respiration with oxygen.
9. If convulsions occur and hypoglycaemia is not a contributing factor, the administration of succinylcholine is encouraged. Do not give CNS depressants such as barbiturates, paraldehyde or morphine.
10. Give phytonadione in cases of abnormal bleeding or hypoprothrobinemia. Fress blood or platelet transfusion may be necessary.
11. If renal function is impaired, dialysis must be used to remove salicylate. If peritoneal dialysis is used, add 5% human albumin (Albuminer) to the dialysate. Potassium chloride, 5meq/L should be added to the dialysate unless serum potassium is elevated.
12. Reduce hyperpyrexia by tepid sponging. Do not use alcohol for sponging. Elevated body temperature should not be allowed to persist.
Detection
Heat 0.3-0.5 g of aspirin for a few minutes with a little sodium hydroxide solution. Cool by setting the test tube in ice water and then acidify the solution using sulphuric acid.
a. Test a portion of the product that will re-crystallize out using Chloride TS
b. The second component of hydrolysis of aspirin is tested by adding to the filtrate 3gtts of alcohol and 5gtts of concentrated sulphuric acid.
SALICYLIC ACID
Synonyms: o-hydroxybenzoic acid
Properties: Salicylic acid occurs as salicylic aldehyde and methyl ester or as a glucoside (salicin). It crystallizes from water in long, white needles having a sweet, acidulous taste and rather harsh taste. It is soluble in about 500 parts of cold water in 13 parts of boiling water (38:1). It is freely soluble in alcohol (1:2), ether (1:2), chloroform, acetone and carbon disulfide. It melts at 157 °C. When heated carefully and slowly in water bath, salicylic acid is in fine needle.
Toxic Action: Salicylic acid produces local effects. Locally it acts like phenol but considerably more feebly than carbolic acid, coagulating albumin and so killing living protoplasm when in very intimate contact. If salicylic acid is applied to mucous surfaces, it produces white eschar as well as inflammation and necrotic sloughing as a result of this action. Absorption of salicylic acid takes place from all mucous surfaces with such rapidity that it can be detected in the urine even after 15 minutes. The effects of absorption appear in the blood vessels, then in the central nervous system and kidneys. They arise from injury to protoplasm as in local action in the kidneys, they result from damage to the secreting epithelium.
Uses: Applied topically over thickened skin (corns) and in dermatological ointments.
Symptoms:
1. Nausea, vomiting and a feeling of fullness in the head
2. Ringing in the ears, dimness of vision
3. Profuse perspiration, confusion and dullness.
4. Irritation of the throat and stomach, leading to vomiting and difficulty in swallowing.
5. Flushing of the face.
6. Dyspnea, weak pulse, subnormal temperature.
7. Convulsion, coma and death in severe cases.
Treatment:
1. Wash out the stomach with sodium bicarbonate (10 grams in 1 pint)
2. Allay burning sensation with milk, eggs or vegetable oil.
Detection
1. Ferric Chloride Test
Add 1mL of aqueous solution of salicylic acid, 5 drops of ferric chloride solution.
2. Millon’s Test
Warm 1mL of aqueous solution of salicylic acid with 10gtts of Millon’s reagent.
3. Jorrisen’s test
Dissolve 1g of salicylic acid in 10mL of water by heating. Cool and then add 5gtts of 10% potassium nitrite, 2gtts of glacial acetic acid and 2gtts of 10% copper sulfate. Boil for 2 minutes.
4. Bromine Water Test
Place 1mL of salicylic acid solution in a test tube and add an excess of freshly prepared bromine water solution.
5. Methyl Ester Test
Warm in a water bath in a mixture of 1mL of salicylic acid with 5gtts of methyl alcohol and 5gtts of concentrated sulphuric acid.
ACETANILIDE
Synonyms: Antifebrin, Acetylated Aniline
Lethal dose: 4-15grains
Properties: It is a colorless, crystalline substance with burning taste but without any odor. It is isolated from an aqueous acid solution by ether or chloroform extraction.
Uses: Analgesic, antipyretic
Toxic Action: As/a derivative of aniline, acetanilide possesses the poisonous properties of aniline though in less degree by reason of the presence of an acetyl group. It behaves like phenacetin although acetanilide is slightly more irritating.
Symptoms:
1. Marked and prolonged cyanosis, great weakness, prostration and collapse.
2. Cold extremities, profuse sweating with feeble, shallow respiration, unconsciousness.
3. Convulsion and delirium
4. Persistent vomiting and dyspnoea.
5. Acute nephritis, suppression of urine (anuria) and acute jaundice.
Treatment:
1. Wash out stomach and administer cathartic.
2. Blood transfusion may be necessary.
Detection
1. Indophenol Test
Boil acetanilide with 1mL of concentrated HCl in a water bath. Evaporate to about 10gtts. Cool and add 1mL of saturated aqueous phenol solution. Add freshly prepared solution of calcium hypochlorite drop by drop. Observe the color formed on shaking. Then carefully add ammonium hydroxide solution as an upper layer. Do not shake. Observe the result.
2. Color Test
One gram of acetanilide is treated with 1mL of concentrated HCl and then diluted with 12mL of water and gently heated to boiling. Cool and divide into equal parts.
a. To one portion add 1mL of fresh aqueous calcium hypochlorite (run along gently at the side of the test tube)
b. To another portion add 1mL of 2% KMnO4 solution.
3. Azo-Dyestuff Test
Boil 1g of acetanilide with 1mL concentrated HCl in a water bath. Cool and place the tube in an ice bath and then diluted with 5mL of distilled water. Add 4gtts of 10%KNO2 make alkaline with NaOH and add a few mg (monggo grain) of beta napthol.
PHENACETIN
Synonyms: Acetophenetidin
Lethal dose: 0.2-1g/Kg
Properties: Phenacetin crystallizes in shining leaflets without color, odor nor taste and melts at 134-135°C. It is soluble in 1400 parts of cold water, in about 70parts of boiling water, 16parts of alcohol and freely soluble in ether and chloroform.
Toxic Action: After absorption of phenacetin it affects principally the brain, temperature regulation and blood. It affects neuralgic pains, nervous headache and irritability. It depresses the motor areas of the cerebral cortex. It lowers temperature by increasing heat elimination via the skin. Phenacetin causes cyanosis by the formation of methemoglobinemia.
Symptoms:
Acute Poisoning:
1. Sweating, gastric irritation, chills and tinnitus.
2. Fall in blood pressure, circulatory collapse, cyanosis, jaundice and oliguria (scanty urine).
3. Coma, convulsions and death due to respiratory failure.
Chronic Poisoning
1. Abdominal pains, headache, cyanosis from methemoglobinemia
2. Hemolytic anemia, weakness, dizziness, irritability
3. Low blood pressure, sleeplessness.
4. Skin eruptions characterized by erythema and popular or ulcerative acne.
5. Severe renal injury characterized by chronic interstitial nephritis with papillary necrosis and severe tubular degeneration which is sometimes irreversible and fatal.
Treatment:
1. Remove ingested drug by emesis with ipecac unless patient is depressed.
2. Activated charcoal interferes with the use of N-acetylcystein.
3. Give saline cathartic like sodium sulfate at 250g /Kg.
4. If the patient is depressed, use airway protected gastric lavage.
5. Keep the patient warm and quiet.
6. Give phytonadione, fresh frozen plasma or clotting factor when necessary for bleeding.
7. Hemodialysis is useful only in the first 10hours.
8. Give 5% dextrose IV for the first 48hours
9. If cyanosis is severe, give methylene blue IV.
Detection
1. Nitration Test (Autenreith-Hinsberg Test)
Heat a monggo grain of phenacetin with 1mL of 10% nitric acid.
2. Oxidation Test
Boil a monggo grain of phenacetin with 1mL of concentrated HCl acid. Dilute with 10mL of distilled water, cool and filter. To the filtrate add 2-3gtts of 3% chromic acid solution. Strong chlorine water may be used instead of chromic acid.
3. Differentiation Test
a. Phosphomolybdic acid is added to separate samples of acetanilide and phenacetin. Heat both phenacetin and acetanilide and observe the results. Use monggo grain amount only.
b. With potassium bromated solution. Dissolve the 2 samples (monggo grain) in concentrated HCl acid, gently boil, then add 5gtts. of potassium bromated solution.
c. With potassium permanganate solution. Boil the 2 samples with 5gtts of HCl acid for 5minutes. Filter and then dilute with an equal amount of water. To the cooled filtrate add 5 gtts. of 10% KMNO4 solution.
4. Indophenol Test
Boil phenacetin with 1mL concentrated HCl in a water bath. Evaporate to about 10gtts. Cool and add 2mL of saturated aqueous phenol solution. Add freshly prepared solution of calcium hypochlorite drop by drop. Observe the color formed on shaking. Then carefully add ammonium hydroxide solution as an upper layer. Do not shake. Observe the result.
5. Azo-dyestuff Test
Boil 1g of phenacetin with 1mL concentrated HCl in a water bath. Cool and place the tube in an ice bath and then diluted with 5mL of distilled water. Add 4gtts. of 10% KNO2 make alkaline with N