Accessed - 1,874 times.

Jaiswal AK1*, Kamna Sharma2, Lukose Sally3, Tabin Millo4
1, 4Department of Forensic Medicine and Toxicology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi-110029, India.
2, 3Department of Forensic Science, Galgotias University, Greater Noida, NCR-Delhi, India.

REVIEW ARTICLE
Volume 2, Issue 3, Page 172-177, September-December 2014.

Article history
Received: 04 October 2014
Revised: 10 November 2014
Accepted: 20 November 2014
Early view: 28 November 2014

*Author for correspondence
E-mail: ashokjaiswal72@gmail.com

ABSTRACT

Antihistamines refer to drugs that antagonize the action of histamines at H1 receptors. It is usually given as tablets for allergic disorders and other conditions such as colds, motion sickness and Parkinson’s disease. In India, Forensic Science Laboratories run by Government under the Home ministry usually carry out the toxicological analysis of these drugs. The samples have to be analyzed by the forensic toxicologist/chemists/scientist. This article deals with the screening/spot test for antihistamines. It attempts to simplify the standard procedures in a step-wise manner, which can be of handy reference for the forensic toxicologist.

Keywords: Antihistamines, Screening, Spot test etc.

INTRODUCTION

Antihistamines are medicines that can be used to relieve severe itching, allergies and their reactions by blocking the action of the histamines on the tissue which are classified as first generation and second generation histamines (Table 1). We have tried to set out standard procedures for screening/spot test for alkaloids which are easily available and useful for the forensic science laboratory. It is in continuation to toxicology manual series article-XVIII, Screening/spot test of Aphrodisiacs (Sex drugs) Vol4, No3, Jan-July 2012 published in International Journal of Medical Toxicology and Legal Medicine. This article covers the spot test/colour test of antihistamines like alimemazine, antazoline, azatadine, bamipine, bromazine, brompheniramine, carbinoxamine, chlorcyclizine, chloropyramine, chlororpyrilene, clemastin,cyproheptedine, deptropine, dimetindene, dimetotiazine, diphenhydramine, histapyrrodine, isothipendyl, mebhydrolin, meclozine, mepyramine, mequitazine, methapyrilene, phenindamine, pheniramine, pizotifen, promethazine, pyrrobutamine, thenalidine, thenyldiamine, thiazinamium metisulfate, thonzylamine, tolpropamine, trimethobenzamide and triprolidine (Modi, 2005; Parikh, 2005; Ng et al. 2004; Ramaekers and Vermeeren, 2000; Lewis, 2014; Antihistamines, 2014; Antihistamines- Side effects; 2014).

Table1. Classification of antihistamines.
Click here to view full image

1. Alimemazine
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet colour is observed which indicates the presence of alimemazine.
Forrest test
1. One to two ml of extract is taken in test tube.
2. Few drops of forrest reagent are added to it.
3. Reddish brown colour is observed which indicates the presence of alimemazine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of alimemazine.
Formaldehyde-sulphuric acid test
1. One to two ml of extract is taken in test tube.
2. Few drops of Formaldehyde-sulphuric acid reagent are added to it.
3. Reddish violet colour is observed which indicates the presence of alimemazine.
2. Antazoline
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of antazoline.
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Red colour is observed which indicates the presence of antazoline.
3. Azatadine
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Pink colour is observed which indicates the presence of azatadine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet colour is observed which indicates the presence of azatadine.
4. Bamipine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Reddish orange colour is observed which indicates the presence of bamipine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Brown colour is observed which indicates the presence of bamipine.
5. Bromazine
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Yellow colour is observed which indicates the presence of bromazine.
6. Brompheniramine
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Orange colour is observed which indicates the presence of brompheniramine.
7. Carbinoxamine
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Orange to pink colour is observed which indicates the presence of carbinoxamine.
8. Chlorcyclizine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Reddish orange colour is observed which indicates the presence of chlorcyclizine.
9. Chloropyramine
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Yellow colour is observed which indicates the presence of chloropyramine.
10. Chloropyrilene
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of chloropyrilene.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet colour is observed which indicates the presence of chloropyrilene.
11. Clemastine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Brown colour is observed which indicates the presence of clemastine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Yellow colour is observed which indicates the presence of clemastine.
Sulphuric acid test
1. Few drops of extract are taken on a white tile.
2. Few drops of sulphuric acid are added to it.
3. Yellow colour is observed which indicates the presence of clemastine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Yellowish brown colour is observed which indicates the presence of clemastine.
12. Cyproheptadine
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Grey to green colour is observed which indicates the presence of cyproheptadine.
Sulphuric acid test
1. Few drops of extract are taken on a white tile.
2. Few drops of sulphuric acid are added to it.
3. Violet colour is observed which indicates the presence of cyproheptadine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of cyproheptadine.
13. Deptroprine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Yellow colour is observed which indicates the presence of deptroprine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Yellow colour is observed which indicates the presence of deptroprine.
14. Dimetindene
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Green colour is observed which indicates the presence of dimetindene.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Blue colour is observed which indicates the presence of dimetindene.
15. Dimetotiazine
Forrest test
1. One to two ml of extract is taken in test tube.
2. Few drops of forrest reagent are added to it.
3. Red colour is observed which indicates the presence of dimetotiazine.
Formaldehyde-sulphuric acid test
1. One to two ml of extract is taken in test tube.
2. Few drops of Formaldehyde-sulphuric acid reagent are added to it.
3. Yellow colour is observed which indicates the presence of dimetotiazine.
FPN test
1. One to two ml of extract is taken in test tube.
2. Few drops of FPN reagent are added to it.
3. Red colour is observed which indicates the presence of dimetotiazine.
16. Diphenhydramine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Orange to brown colour is observed which indicates the presence of diphenhydramine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Yellow colour is observed which indicates the presence of diphenhydramine.
Sulphuric acid test
1. Few drops of extract are taken on a white tile.
2. Few drops of sulphuric acid are added to it.
3. Orange colour is observed which indicates the presence of diphenhydramine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Yellow colour is observed which indicates the presence of diphenhydramine.
17. Histapyrrodine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Reddish violet colour is observed which indicates the presence of histapyrrodine.
18. Isothipendyl
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Orange colour is observed which indicates the presence of isothipendyl.
19. Mebhydrolin
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Blue colour is observed which indicates the presence of mebhydrolin.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Greyish blue colour is observed which indicates the presence of mebhydrolin.
20. Meclozine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Reddish brown colour is observed which indicates the presence of meclozine.
21. Mepyramine
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Yellow colour is observed which indicates the presence of mepyramine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of mepyramine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet colour is observed which indicates the presence of mepyramine.
22. Mequitazine
Forrest test
1. One to two ml of extract is taken in test tube.
2. Few drops of forrest reagent are added to it.
3. Red colour is observed which indicates the presence of mequitazine.
Formaldehyde-sulphuric acid test
1. One to two ml of extract is taken in test tube.
2. Few drops of Formaldehyde-sulphuric acid reagent are added to it.
3. Reddish violetcolour is observed which indicates the presence of mequitazine.
FPN test
1. One to two ml of extract is taken in test tube.
2. Few drops of FPN reagent are added to it.
3. Red colour is observed which indicates the presence of mequitazine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Red colour is observed which indicates the presence of mequitazine.
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Green colour is observed which indicates the presence of mequitazine.
23. Methapyrilene
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet to black colour is observed which indicates the presence of methapyrilene.
Sulphuric acid test
1. Few drops of extract are taken on a white tile.
2. Few drops of sulphuric acid are added to it.
3. Orange colour is observed which indicates the presence of methapyrilene.
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Brown colour is observed which indicates the presence of methapyrilene.
24. Phenindamine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Red colour is observed which indicates the presence of phenindamine.
25. Pheniramine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Reddish orange colour is observed which indicates the presence of pheniramine.
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Orange colour is observed which indicates the presence of pheniramine.
26. Pizotifen
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Black colour is observed which indicates the presence of pizotifen.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet to green colour is observed which indicates the presence of pizotifen.
Sulphuric acid test
1. Few drops of extract are taken on a white tile.
2. Few drops of sulphuric acid are added to it.
3. Orange to yellow colour is observed which indicates the presence of pizotifen.
27. Promethazine
Forrest test
1. One to two ml of extract is taken in test tube.
2. Few drops of forrest reagent are added to it.
3. Red colour is observed which indicates the presence of promethazine.
FPN test
1. One to two ml of extract is taken in test tube.
2. Few drops of FPN reagent are added to it.
3. Orange colour is observed which indicates the presence of promethazine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet to green colour is observed which indicates the presence of promethazine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Violet colour is observed which indicates the presence of promethazine.
28. Pyrrobutamine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Brown colour is observed which indicates the presence of pyrrobutamine.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet colour is observed which indicates the presence of pyrrobutamine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Grey to violet colour is observed which indicates the presence of pyrrobutamine.
29. Thenalidine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Orange to brown colour is observed which indicates the presence of thenalidine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Grey to violet colour is observed which indicates the presence of thenalidine.
30. Thenyldiamine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Green colour is observed which indicates the presence of thenyldiamine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Black to violet colour is observed which indicates the presence of thenyldiamine.
31. Thiazinamium metisulfate
Forrest test
1. One to two ml of extract is taken in test tube.
2. Few drops of forrest reagent are added to it.
3. Red colour is observed which indicates the presence of thiazinamium metisulfate.
FPN test
1. One to two ml of extract is taken in test tube.
2. Few drops of FPN reagent are added to it.
3. Red colour is observed which indicates the presence of thiazinamium metisulfate.
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Red colour is observed which indicates the presence of thiazinamium metisulfate.
32. Thonzylamine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Reddish violet colour is observed which indicates the presence of thonzylamine.
33. Tolpropamine
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Brown colour is observed which indicates the presence of tolpropamine.
Marquis test
1. One to two ml of extract is taken in test tube.
2. Few drops of marquis reagent are added to it.
3. Red colour is observed which indicates the presence of tolpropamine.
34. Trimethobenzamide
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Black colour is observed which indicates the presence of trimethobenzamide.
Mandelin’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of mandelin’s reagent are added to it.
3. Violet to brown colour is observed which indicates the presence of trimethobenzamide.
35. Triprolidine
Libermann’s test
1. One to two ml of extract is taken in test tube.
2. Few drops of libermann’s reagent are added to it.
3. Orange colour is observed which indicates the presence of triprolidine.
Cyanogen bromide test
1. One to two ml of extract is taken in test tube.
2. Few drops of cyanogens bromide reagent are added to it.
3. Orange to pink colour is observed which indicates the presence of triprolidine.
Preparation of Solutions/Reagents
1. Cyanogen bromide: (I) Decolourisation of bromine water is done by the addition of solid potassium cyanide and then more bromine solution is added until the solution becomes pale yellow. (II) Prepare a saturated solution of aniline in water. Mix the solution (I) and (II).
2. Formaldehyde-sulphuric acid: Four volumes of sulphuric acid and six volumes of formaldehyde solution are mixed.
3. Forrest reagent: Equal volumes of a 0.2 % (w/v) solution of potassium dichromate, 30 % (w/v) solution of sulphuric acid, 20 % (w/v) solution of perchloric acid and 50 % solution of nitric acid are mixed.
4. FPN reagent: 5 ml of 5 % (w/v) ferric chloride solution, 45 ml of 20 % (w/w) solution of perchloric acid and 50 ml of 50 % (v/v) solution of nitric acid are mixed.
5. Libermann’s reagent: 1 gm of sodium or potassium nitrite is mixed in 10 ml of sulphuric acid with cooling and swirling to absorb the brown fumes.
6. Mandelin’s reagent: 1 g of ammonium vanadate is dissolved in 1.5 ml of water and diluted to 100 ml with concentrated sulphuric acid.
Marquis reagent: 100 ml of concentrated sulphuric acid is mixed with 1 ml of 40% (v/v) formaldehyde solution.

CONCLUSION

In any analysis of poison, screening/spot test is very useful for knowing the presence of the anti-histamines which can be confirmed by the more confirmatory tests. It saves time for the toxicologist in ruling out the poisons which can be confirmed by the more confirmatory tests. It saves time for the toxicologist in ruling out the 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. It is important for the forensic toxicologists to know the best available method and help to detect the poison in the crime investigations.

CONFLICT OF INTEREST
None declared.

REFERENCES

Antihistamines- Side effects [www.nhs.uk/Conditions/Antihistamines/Pages/Side-effects.aspx]. Accessed on 20-September-2014.
Antihistamines. [http://www.users.globalnet.co.uk/~aair/antihistamines.htm#WHATRANTI]. Accessed on 25-September-2014.
Lewis V. Antihistamines.
[http://www.netdoctor.co.uk/skin_hair/eczema_antihistamines_003764.htm]. Accessed on 12-September-2014.
Modi JP. Modi’s textbook of Medical Jurisprudence and Toxicology. 23rd edition. Lexis-Nexis: India. 2005.
Ng KH, Chong D, Wong CK, Ong HT, Lee CY, Lee BW, Shek LP. Central nervous system side effects of first- and second-generation antihistamines in school children with perennial allergic rhinitis: a randomized, double-blind, placebo-controlled comparative study. Pediatrics. 113, 116-121, 2004.
Parikh’s Textbook of Medical Jurisprudence, Forensic Medicine & Toxicology, 6th edition. CBS Publishers & Distributors: New Delhi. 2005.
Ramaekers JG, Vermeeren A. All antihistamines cross blood-brain barrier. BMJ. 321, 572, 2000.