Screening/spot test of narcotics
Introduction
Narcotics, the word is borrowed from the Greek word ‘narkōtikós’, which means lethargic or stuporous. The expression narcotic is generally used to refer to a class of drugs that are analgesics, central nervous system depressants and produces insensibility or stupor. Narcotics commonly termed as Opioids, usually produces analgesia, mood swings, physiological dependence, heavy tolerance and a hedonic or rewarding effect, which contributes to compulsive use of the drug by the abuser. The effects caused by narcotics include euphoria, drowsiness, respiratory, depression, constipation, constricted pupils. Narcotics majorly attacks the Central Nervous System and Peripheral Nervous System and usually found in these areas as well as in the Gastrointestinal Tract. Such drugs act by binding to opioid receptors in the body, as narcotics mediate both the psychoactive and somatic effects of opioids, though they do not cross the blood-brain barrier, but can shift other opioids from binding to the respective receptors. Forensic Scientists/Toxicologists have a hard time in searching appropriate tests to analyse samples of narcotics in medico-legal cases, autopsies, and even for academic purposes.1, 2, 3, 4, 5, 6, 7, 8, 9
In India, The Narcotic Drugs and Psychotropic Substance (NDPS) Act, 1985 covers all kinds of activities related to narcotic substances. Bare act reads as follows:
“An Act to consolidate and amend the law relating to narcotic drugs, to make stringent provisions for the control and regulation of operations relating to narcotic drugs and psychotropic substances [to provide for the forfeiture of property derived from, or used in, illicit traffic in narcotic drugs and psychotropic substances, to implement the provisions of the International Conventions on Narcotic Drugs and Psychotropic Substances] and for matters connected therewith.”10
In simplified terms, the act states that possessing, manufacturing, production, selling, transportation, purchasing, inter-state importing and exporting, or using cannabis is punishable with rigorous imprisonment attracting ten years of jail or fine extending up to one lakh rupees.
We have tried to set out standard procedures for screening/spot test for narcotics that are easily available and useful for the forensic science laboratory. This article covers the different narcotics drugs like Acetorphine, Alphaprodine, Anileridine, Benzylmorphine, Bezitramide, Buprenorphine, Butorphanol, Codeine, Cyclazocine, Dextromoramide, Dextropropoxyphene, Diamorphine, Diethylthiambutene, Dihydrocodeine, Dipipanone, Embutramide, Etorphine, Ketobemidone, Methadone, Morphine, Pentazocine, Pethidine, Phenazocine, Phenoperidine, Profadol and Thebacon.
Acetorphine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
The bluish-grey colour is observed which changes to yellowish-brown indicate the presence of acetorphine.
Alphaprodine
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
The bluish-grey colour is observed which indicates the presence of alphaprodine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
The brownish-red colour is observed which indicates the presence of alphaprodine.
Anileridine
Diazotisation test
Few drops of the extract are dissolved in 2M hydrochloride acid.
One drop of it is then taken on a tile.
One drop of 1% solution of sodium nitrite followed by a drop of 4% solution of naphtha-2-ol in 2M sodium hydroxide is added to it.
Orange colour is observed which indicates the presence of anileridine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Orange colour is observed which indicates the presence of anileridine.
Benzylmorphine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
The red colour is obtained which changes to violet indicate the presence of benzylmorphine.
Bezitramide
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Violet colour is observed which changes to orange shows the presence of bezitramide.
Buprenorphine
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of buprenorphine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Violet colour is observed which indicates the presence of buprenorphine.
Butorphanol
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of butorphanol.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Grey colour is observed which indicates the presence of butorphanol.
Codeine
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of codeine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Violet colour is observed which indicates the presence of codeine.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green colour is observed which changes to orange shows the presence of codeine.
Cyclazocine
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of cyclazocine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Brown colour is observed which changes to green indicate the presence of cyclazocine.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green colour is observed which changes to orange shows the presence of cyclazocine.
Dextromoramide
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
The solution is heated at 100℃.
Green colour is observed which indicates the presence of dextromoramide.
Dextropropoxyphene
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Brown colour is observed which indicates the presence of dextropropoxyphene.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Play of colour is observed which changes from black to violet and finally to green which indicates the presence of dextropropoxyphene.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Grey to brown colour is observed which shows the presence of dextropropoxyphene.
Diamorphine
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of diamorphine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Violet colour is observed which indicate the presence of diamorphine.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Blue to grey colour is observed which shows the presence of diamorphine.
Diethylthiambutene
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
The solution is heated at 100℃.
Orange colour is observed which indicates the presence of diethylthiambutene.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Violet colour is observed which indicate the presence of diethylthiambutene.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green to greenish-blue colour is observed which shows the presence of diethylthiambutene.
Sulphuric acid test
Two ml of extract is taken in a test tube.
Few drops of sulphuric acid are added to it.
Orange colour is observed which indicate the presence of diethylthiambutene.
Nitric acid test
Two ml of extract is taken in a test tube.
Few drops of cold nitric acid are added to it.
A pink-brown colour change to green is observed which indicate the presence of diethylthiambutene.
Dihydrocodeine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Violet colour is observed which indicate the presence of dihydrocodeine
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Grey to green colour is observed which shows the presence of dihydrocodeine.
Dipipanone
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green to blue colour is observed which shows the presence of dihydrocodeine.
Embutramide
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Blue colour is observed which indicate the presence of embutramide.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Brown colour is observed which shows the presence of embutramide.
Etorphine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
The bluish-grey colour to yellowish-brown is observed which indicate the presence of etorphine.
Ketobemidone
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Orange colour is observed which indicate the presence of ketobemidone.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Blue to green colour is observed which shows the presence of ketobemidone.
Methadone
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Brown to orange colour is observed which indicates the presence of methadone.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green to blue colour is observed which shows the presence of methadone.
Morphine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Play of colour from purple-red which gradually changes to violet and finally to blue which shows the presence of morphine.
Froehde’s test
A drop of the extract is taken in a porcelain basin.
The solution is then dried.
Few drops of Froehde’s reagent are added to it.
Play of colour from violet changes to green and finally to pink colour which shows the presence of morphine.
Ferric chloride test
Two ml of extract is taken in a test tube.
Few drops of ferric chloride solution are added to it.
Blue colour is observed which shows the presence of morphine.
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of morphine.
Porphyroxine test
Two ml of the extract is taken in a test tube.
Few drops of acetic acid followed by dilute hydrochloric acid are added to it.
Pink or rose red colour is obtained which indicates the presence of morphine.
Pentazocine
Folin-Ciocalteu test
Two ml of extract is taken in a test tube.
Few drops of folin-ciocalteu reagent are added to it.
The solution is made alkaline with 2M sodium hydroxide.
Blue colour is obtained which indicates the presence of pentazocine.
Mandelin’s test
Two ml of extract is taken in a test tube.
Few drops of mandelin’s reagent are added to it.
Green colour is observed which shows the presence of pentazocine.
Pethidine
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Reddish orange colour is observed which indicates the presence of pethidine.
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Orange colour is observed which indicate the presence of pethidine.
Phenazocine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Orange colour is observed which indicate the presence of phenazocine.
Phenoperidine
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Red colour is observed which indicate the presence of phenoperidine
Profadol
Folin-Ciocalteu test
Two ml of extract is taken in a test tube.
Few drops of folin-ciocalteu reagent are added to it.
The solution is made alkaline with 2M sodium hydroxide.
Blue colour is obtained which indicates the presence of profadol.
Liebermann’s test
Two ml of extract is taken in a test tube.
Few drops of libermann’s reagent are added to it.
Black colour is observed which indicates the presence of profadol.
Thebacon
Marquis test
Two ml of extract is taken in a test tube.
Few drops of marquis reagent are added to it.
Yellow to violet colour is observed which indicate the presence of thebacon.
Conclusion
During the analysis of any poison, screening/spot test is very useful for knowing the presence of the narcotics by such preliminary tests, which can be confirmed by the confirmatory tests later on. Thus, it saves time for the toxicologist in ruling out various non-narcotic poisons and gives a quick clue to the doctors for patient management in emergency poisoning cases. The result of the analytical methods depends on the amount and purity of the sample extracted. Screening/spot test has been developed after repeated trial and testing. The techniques are being improved every time. The forensic toxicologists need to know the best available method and help to detect the poison in criminal investigations.
Preparation of Solutions
Ferric chloride solution: 10 g of ferric chloride is dissolved in 100 ml distilled water.
Folin-Ciocalteau reagent: Dissolve 100 g of sodium tungstate and 25 g of sodium molybdate in 800 ml of purified water in a 1.5-l flask. Add 50 ml of concentrated orthophosphoric acid and 100 ml of concentrated hydrochloric acid and reflux for 10 hours. Cool, add 150 g of lithium sulfate, 50 ml of purified water and 0.5 ml of elemental bromine, and allow to stand for 2 hours. Boil for 15 minutes to remove excess bromine, cool, filter if necessary, and dilute to 1 litre with purified water. This solution is yellow and should be stable for 4 months if stored at 4°C.
Froehde’s reagent: 1 g of molybdic acid or sodium molybdate is dissolved in 100 ml of hot concentrated sulphuric acid.
Liebermann’s reagent: 1 gm of sodium or potassium nitrite is dissolved in 10 ml of sulphuric acid with cooling and swirling to absorb the brown fumes.
Mandelin’s reagent: 1 g of ammonium vanadate is dissolved in 1.5 ml of water and dilute to 100 ml with concentrated sulphuric acid.
Marquis reagent: 100 ml of concentrated sulphuric acid is mixed with 1 ml of formaldehyde solution. [40% (v/v)].
Conflict of Interest
None.