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Modified Scott's test for ketamine

hydrochloride
a

Pallavi Dubey , Sudhir K. Shukla & Kailash C. Gupta

CFSL, Chemistry , Plot no. 2, Dakhin Marg, Sector 36A,

Chandigarh , 160036 , India
b

Central Forensic Science Laboratory, Ministry of Home Affairs ,

Ramanthapur , Hyderabad , 500013 , India
c

IIT, Chemistry, Indian Institute of Technology , Roorkee ,

247667 , India
Published online: 26 Oct 2012.

To cite this article: Pallavi Dubey , Sudhir K. Shukla & Kailash C. Gupta (2013) Modified Scott's
test for ketamine hydrochloride, Australian Journal of Forensic Sciences, 45:2, 165-171, DOI:
10.1080/00450618.2012.733026
To link to this article: http://dx.doi.org/10.1080/00450618.2012.733026

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Australian Journal of Forensic Sciences, 2013

Vol. 45, No. 2, 165171, http://dx.doi.org/10.1080/00450618.2012.733026

Modied Scotts test for ketamine hydrochloride

Pallavi Dubeya*, Sudhir K. Shuklab and Kailash C. Guptac

Downloaded by [The University of Manchester Library] at 06:29 12 October 2014

CFSL, Chemistry, Plot no. 2, Dakhin Marg, Sector 36A, Chandigarh, 160036 India; bCentral
Forensic Science Laboratory, Ministry of Home Aairs, Ramanthapur, Hyderabad, 500013
India; cIIT, Chemistry, Indian Institute of Technology, Roorkee, 247667, India
(Received 3 June 2012; nal version received 17 September 2012)
Ketamine is a drug of abuse and is gaining popularity as a date rape drug. There
are no eld test kits available especially for ketamine. Scotts test has been used
for the detection of cocaine for a very long time. It is based upon the formation of
an insoluble complex of the cobalt thiocyanate ion with a free nitrogen base of
cocaine. However, this test has been known for a large number of false positives
when tested for cocaine. The authors have modied Scotts test by basifying the
original solution and the addition of ethylene glycol for the detection of ketamine
hydrochloride in the presence of lignocaine and cocaine. The test was done on
more than 100 common drugs of abuse, medicines, cutting agents, excipients and
other substances and was found to be specic only for ketamine. The EDX and
FTIR analysis was further carried to establish the presence of ketamine in the
complex. There were no false positives for the test and hence it could be
incorporated for routine drug screening in toxicological and forensic drug
laboratories, as well as frontline drug enforcement agencies.
Keywords: Ketamine Hydrochloride, Scotts test, EDX

1. Introduction
Ketamine hydrochloride or 2-(2-chlorophenyl)-2-(methylamino) cyclohexanone
hydrochloride is widely used as an anaesthetic drug that stimulates NMDA (Nmethyl-D-aspartate) receptors on neuronal cells1. Inhibition of potassium channels,
especially in central neurons, has been suggested to underlie some of the excitatory
eects and emergence phenomena observed with a dissociative anaesthetic such as
ketamine2. Analytical techniques used in analysis include infrared spectroscopy3, gas
chromatography/mass spectrometry (GC/MS)4 and ultraviolet spectroscopy58.
Currently, only the Janovsky colour test has been reported for presumptive
ketamine detection9. However, several benzodiazepines give the same colour with the
Janovsky reagent. A presumptive test for ketamine hydrochloride was reported by
Morris in 2007. The test is a modication of the standard cobalt thiocyanate
(CoSCN)2 test for cocaine and was specic for ketamine hydrochloride. The
drawbacks of the test included positive results for mepivacaine, phencyclidine and
many other drugs of abuse10. Another method for a spot test was suggested by

*Corresponding author. Email: d_pallavi18@yahoo.com

2012 Australian Academy of Forensic Sciences

P. Dubey et al.

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166
2

Sarvar11 using gold bromide solution, but that gave the same results with morphine
and the reagents were not available in all laboratories.
A new presumptive test for the determination of ketamine is reported below. The
authors have prepared the tetrathiocyanates of cobalt [Co(SCN)4]- is an alternative
to the Co(SCN)2 which gives the same reaction with lidocaine, cocaine and several
other basic drugs. The use of ketamine as a date rape drug and a popular drug of
abuse is increasing in Asia. The test enables rapid screening and detection of
ketamine without any false positive results and hence, prior doing any sophisticated
instrumentation, ketamine would be fairly detected in the matrices in pure and
adulterated forms. The test is straightforward, denitive, and utilizes reagents
commonly used in forensic drug analysis. The drug test kits can eciently use this
reagent for the detection of ketamine. In addition, if the solution is not basied, it
could be used as a conventional test for cocaine.
2. Experimental
2.1. Materials and methods
Ketamine hydrochloride was supplied by Neon Laboratories Pvt. Ltd. Nickel
chloride, ammonium thiocyanate and ethylene glycol were all Merck India products.
The standard reference materials for NDPS drugs and pharmaceutical products were
obtained from the Department of Chemistry, Central Forensic Science Laboratory,
Government of India.
2.2.

Preparation of solutions

Preparation of cobalt tetrathiocyanate complex: 0.29 g of cobalt chloride was

dissolved in 100 ml of water and heated for 15 minutes. 0.304 g of ammonium
thiocyanate was taken in 100 ml water and was boiled for 20 minutes; 50 ml of each
solution was taken and mixed .The resultant solution was heated for 2 hours and the
crystals obtained at the bottom of the beaker were taken out in a watch glass and
were dried over a water bath. The resultant Prussian blue crystals were stored in a
dried vial in a dessicator for further use. A 0.1 M aqueous solution of cobalt
tetrathiocyanate was prepared and was further diluted with ethylene glycol in a 1:1
ratio. The solution was stored at 48C for further use.
2.3. Test for ketamine hydrochloride and narcotic drugs and psychotropic substances
A solid sample of 1 mg was added to the spot plate. For liquid samples, a single drop
of the sample was added to a spot well or a test tube, followed by a single drop of
0.1 N KOH solution. A drop of the cobalt tetrathiocyanate solution was added.
Samples containing ketamine hydrochloride formed purplish blue precipitate. A
negative result was indicated by no change in the original colour of the solution/drug
or a blue solution. A negative test was also performed by side-by-side comparisons of
the blank and the sample-coloured precipitates.
2.4.

Characterization of the complexes

The IR-absorption spectra were obtained by applying the KBr-disc technique using a
Perkin Elmer Fourier-Transform Infrared Spectrophotometer. The KBr used was of

Australian Journal of Forensic Sciences

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analytical grade for the determination of the drug-metal thiocyanate complex. The
SEM EDX analysis was done with a Carl Zeiss EVO MA15 SEM-EDXA System.
The ketamine samples were coated by loading the palladium sputter target and a
pressure of 2 6 102 millibar. The plasma was set and the coating was done for 6 to
7 minutes. For SEM the EHT was set to 20 kV with a beam current of 100 mA and
probe current of 50100 pA. The images were further analysed for elements by using
the INCA software and adjusting the vacuum to a lower value. The qualitative and
quantitative analysis of elements was done by the EDX.
3.

Result and discussion

The addition of ethylene glycol and KOH solution made the reagent specic and
non-reactive to cocaine, lidocaine, mepivacaine, bupivacaine and other drugs, which
is a major source of false positive tests in the eld.
3.1.

Specicity

Eighty compounds covering a wide variety of controlled substance classes,

adulterants, diluents, and excipients were screened using the modied CoSCN test.
Table 1 lists the compounds tested and their individual results. The modied CoSCN
test revealed specicity to ketamine hydrochloride. Precipitates from ketamine
hydrochloride were collected and characterized on EDX and FTIR (Table 2). The
results of elemental analysis for C, H, N, S and metal content are in good agreement
with those required by the suggested formulae, i.e. formation of 2:1 ketamine
Co(SCN)4 ion-associate. The structure of the complex and the pure drug were
observed by SEM and elemental analysis was done by EDX to determine the
empirical formula of the complexes and pure drug. The FTIR spectrums for pure
ketamine and the ketamine-cobaltithiocyanate complex are shown in Figures 1 and 2.
3.2.

Mechanism of the reaction

The preparation of ion associates depends on the mechanism of the binding of cobalt
(II) complexes with the thiocynates to form an octahedral complex [Co (SCN)4]27.
Further, the substitution of two bulky, relatively hydrophobic protonated anaesthetic cations in the coordination sphere lends stability to the complex. The ketamine
cobalttetrathiocyanate was analysed by FTIR to determine the presence of ketamine
and the presence of the thiocyanate bond. The OH ions in the reaction are
important as pH plays a vital role in the formation of the ion association complex.
The proposed mechanism of the formation of complex is as follows:
CoSCNH2 O5  aq 3SCN aq $ CoSCN4 2 aq 5H2 O


CoSCN4

2

aq 2R3 NH KOH $ R3 NH2 CoSCN4

Ketamine

Complex

The elemental analysis conrmed the empirical formula of cobalt tetrathiocyanate and the ketamine cobalttetrathiocyanate complex. FTIR conrmed the presence
of a distinct peak of thiocyanate at 2017 cm71.

P. Dubey et al.

168
4
Table 1.

Tests of the reagent with seventy NDPS drugs and excipients.

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Compound

Test result

Compound

Morphine
Heroin

NR
NR

Dextrose
Diphenhydramine HCl

Codeine
Strychnine
Brucine
Betamethasone
Dexamethasone
Digitoxigenin,
Digoxigenin,

NR
NR
NR
NR
NR
NR
Blue ppt in
pink soln
NR
NR
NR
NR
NR
NR
NR

Ephedrine HCl
Ethylmorphine
Fenuramine HCl
Ketamine
Procaine
Bupivacaine
Mepivacaine

Digitoxin
Lanatoside C
Alprazolam
Amobarbital
Amphetamine sulfate
Citric acid
Cocaine base
Cocaine HCl
Codeine
Ibogaine HCl
Ketamine free base
Ketamine HCl

Lidocaine
Lidocaine HCl
Pethidine
Flunitrazepam
Phenylepropanolamine HCl
Flurazepam 2HCl
butyrolactone (GBL)

Lactose
Levorphanol tartrate
Methamphetamine HCl

NR
NR
NR
NR
Pinkish
purple ppt
NR
NR
NR

Meperidine HCl
Mescaline HCl
Methadone HCl

Methaqualone HCl

NR

Phencyclidine HCl

Methocarbamol
Nicotinamide
Nordiazepam
Opium
Oxazepam

NR
NR
NR
NR
NR

Phenobarbital
Phenylepropanolamine HCl
Quazepam
Secobarbital, Na
Sodium bitartrate

Pentazocine HCl

NR

Sodium carbonate

Triazolam
Saline water
Mannitol
Methyl paraben
1,4-Butanediol

NR
Pink solution
NR
NR
NR

Ethylene glycol

NR

2,4,6-Trimethoxyamphetamine
HCl
2,5-Dimethoxy-4bromoamphetamine HBr
2,5-Dimethoxy-4ethylamphetamine

NR

Soluble starch
Adrenaline
KOH solution
Ethanol
5-Methoxy- amethyltryptamine HCl
5-Methoxy-N,Ndimethyltryptamine
Modanil

NR
NR

GHB, Na
Hydromorphone HCl
Mannitol
MDMA HCl
Medazolam

N,N-Diisopropyl-5methoxytryptamine HCl
N,N-Dimethylamphetamine
HCl

Test result
NR
Bright
blue ppt
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
NR
Pink
solution
NR
NR
NR
NR
NR
NR
NR
Bright
blue ppt
Bright
blue ppt
NR
NR
NR
NR
Pink
solution
Light
blue ppt
NR
NR
NR
NR
NR
NR
NR
NR
NR

Ketamine-cobalttetra
thio cyanate complex

Ketamine HCl

Cobalt Tetrathio-cyanate

EDX spectra

Atomic Mass
12.010
14.006
15.999
35.453
1.008
58.933
32.066

Carbon
Nitrogen
Oxygen
Chlorine
Hydrogen
Cobalt
Sulphur

12.010
14.006
15.999
35.453
1.008

Carbon
Nitrogen
Oxygen
Chlorine
Hydrogen

Element

Atomic Mass

12.010
58.933
14.006
32.066

Carbon
Cobalt
Nitrogen
Sulphur

Element

Atomic Mass

Element

17
5
1
2
16
1
4

No. of atoms

13
1
1
2
17

No. of atoms

4
1
4
4

No. of atoms

Elemental composition

38.59
13.23
3.02
6.70
3.04
11.14
24.24

Mass %

65.68
5.891
6.729
14.91
6.78

Mass %

16.49
20.23
19.24
44.04

Mass %

EDX spectra and elemental composition of ketamine hydrochloride, cobalt tetrathiocyanate and drug-cobalt tetrathiocyanate complex.

Compound

Table 2.

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170
6

P. Dubey et al.

Figure 1.

Ketamine standard FTIR spectra.

Figure 2.

Ketamine cobaltithiocyanate complex FTIR spectra.

The substitution of ethylene glycol by glycerol facilitated the selectivity of the

reagent to ketamine and upon reaction with lidocaine, mepivacaine, bupivacaine and
phencyclidine; no change in colour was obtained. However, no mechanism of
reaction could be explained for the same.

Australian Journal of Forensic Sciences

4.

171
7

Conclusion

A modied CoSCN presumptive colour test for ketamine hydrochloride was

presented. The test has good specicity. Of all the compounds tested in this study,
only ketamine hydrochloride gave a pinkish-purple precipitate. The test is suciently
sensitive enough for analysis of commercial ketamine hydrochloride products. The
already-established Scotts test could be modied by this method and could be used
for the detection of ketamine as there are no spot tests available for determination of
ketamine hydrochloride.

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