Nov 21, 2011 ... Keywords: Glue; Toluene; Gas chromatography-mass spectroscopy detector;
Head ... contact adhesives. The principal ... GC-MSD head space utilized 10 µL
head space Hamilton syringe ... They are Dunlop glue, Good Mark.
Yacob et al., J Drug Metab Toxicol 2011, 2:3 http://dx.doi.org/10.4172/2157-7609.1000112
Drug Metabolism & Toxicology Research Article Research Article
Open OpenAccess Access
Detection of Vapour Metabolites of Glue Sniffer’s Urine Using Head Space Gas Chromatography Mass Spectrometry Abdul Rahim Yacob*, Mohd. Raizul Zainalibdin, Nazirah Said Chemistry Department, Faculty of Science, University Teknologi Malaysia, 81310, Johor Bahru, Johor, Malaysia
Abstract The abuse of volatile substances has been around for many years. The negative and sometimes fatal conclusion of the abuser led to law being regulated but the problem had spiraled almost out of control due to the ease of availability of the materials abused. The abused substances are most commonly found in products of everyday use such as glue, paint and thinner. In Malaysia, the term ‘glue sniffing’ is given to the abuser as they inhale glue vapour that gives them the ‘high’ sensation most drug abusers are seeking for. This study employs gas chromatographymass spectrometry with head space method for it is the vapour that is inhaled by the abuser, thus during secretion in urine, vapour metabolites are expected. Samples for the study are 4 commercially available glues that are reported as being the most commonly abused as well as the urine samples of probable glue sniffers taken during a raid by the Malaysia Anti Drug Agency, Johor branch. In this study, toluene was shown as the major solvent compound found in glue, and metabolites of toluene, namely p-cresol and N-(methylbenzoyl)-methyl ester-glycine were detected in urine samples of those who are positive for glue sniffing.
Keywords: Glue; Toluene; Gas chromatography-mass spectroscopy detector; Head space; Hippuric acid
Introduction For many years, the problem of volatile substance abuse has been around. These substances have the potential to cause major physiological and neurological damage that, in many cases, cannot be remedied. Furthermore, the severe physical damage associated with inhalant abuse has been documented extensively. These volatile substances being inhaled by abusers are recorded worldwide which led to regulations and law being implemented but to no avail in controlling the act from getting any worse [1]. In Malaysia, the abuse of organic volatile solvents has been observed since the early 1980s. The problem of solvent abuse is predominant in East Malaysia (i.e., Sabah and Sarawak) and in Johore the southern part of West Malaysia bordering Singapore [2]. The substances abused include paint thinner, nail polish remover, gasoline and the most frequently abused type of inhalant being glue. The types of glue most often abused frequently have an organic solvent which is toluene [3]. This solvent has a boiling point lower than room temperature. The solvent in glue have a long term negative health effect when inhaled. After inhalation, the vapor gives the user the delusional feeling which will slowly lead to a general feeling of hallucination or ‘high’. In some respects, solvent in glue is even more abused than drugs. This is because glue is generally easier to get and cheaper than drugs [4]. Toluene is the solvent with the most documentation of abuse, possibly because of its relative low risk of sudden death and the ease of detection in blood. It is found in many products including paint and contact adhesives. The principal metabolite of the toluene is benzoic acid (approximately 80% of dose) which is conjugated with the glycine to form hippuric acid for excretion in urine (half-life of 2-3 hour) [5]. The inhalation of toluene in glue by glue sniffers in Malaysia is a growing problem. The exact number or documentation of the number of glue sniffers is not available as glue or solvent abuse is considered as an alternative to drugs, thus it is not reported to the national registry of abusers in Malaysia. A gleam future awaits the country if the problem J Drug Metab Toxicol ISSN: 2157-7609 JDMT, an open access journal
is not addressed correctly and swiftly from now on as it is feared that it will be out of hand when the responsible enforcement agencies could not stop it anymore [6]. Furthermore, there is no law that in Malaysia that is exclusively regulated for these abusers currently [3]. As the solvents in the glue inhaled are volatile substances, head space method may be the best method in ‘capturing’ what exactly is the glue sniffers are inhaling. In this study, the most commonly abused glue types are identified and the properties of the glue are determined using gas spectrometer mass spectrometry using head space method. Blood and urine samples of glue sniffers are also being studied via the same method. This project has considerable potential and the positive impact of its findings to Malaysia and the society can be high. The results of this project will go a long way towards helping the Royal Police and National Anti-Drug agency to prevent this problem from becoming worse.
Materials and Methods Instrumentation GC-MSD head space utilized 10 µL head space Hamilton syringe from Agilent Technologies, USA with a HP-5 capillary column of dimension 30 m x 0.30 µm x 0.30 mm brand J&W Scientific, USA. An isothermal condition employed with 65ºC for 45 minutes, initial oven temperature at 150ºC, injector temperature was set at 150ºC, detector temperature at 200ºC and helium flow rate of 0.7 mL/min [7].
*Corresponding author: Prof. Dr. Abdul Rahim Yacob, Chemistry Department, Faculty of Science, University Teknologi Malaysia, 81310, Johor Bahru, Johor, Malaysia, Tel:+607 5534505; Fax:+607 5566162; E-mail:
[email protected] Received April 19, 2011; Accepted November 18, 2011; Published November 21, 2011 Citation: Yacob AR, Zainalibdin MR, Said N (2011) Detection of Vapour Metabolites of Glue Sniffer’s Urine Using Head Space Gas Chromatography Mass Spectrometry. J Drug Metab Toxicol 2:112. doi:10.4172/2157-7609.1000112 Copyright: © 2011 Yacob AR, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Volume 2 • Issue 3 • 1000112
Citation: Yacob AR, Zainalibdin MR, Said N (2011) Detection of Vapour Metabolites of Glue Sniffer’s Urine Using Head Space Gas Chromatography Mass Spectrometry. J Drug Metab Toxicol 2:112. doi:10.4172/2157-7609.1000112
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Local glue sample selection
Abundance
After an interview session conducted on more than 50 respondents, four different glue samples were selected as the sample for the research. Based on the result of the interview, these four glues brands were the most reported being abused. They are Dunlop glue, Good Mark CA glue, GP glue and Wessbond glue (shoe glue). All the glues were purchased at hardware shops around Taman Universiti, Skudai, Johor Bahru, Johor.
TIC: SAMPLE 9.D\data.ms 19000 18000
15.921
17000 16000 15000 14000 13000 12000
Urine samples
18.146
11000 10000
Urine samples from Sultanah Aminah Hospital, Johor Bahru and three secondary schools in Johor Bahru area were obtained during a raid with the co-operation of the National Anti Drug Agency. All of the urine samples obtained were then screened using G.S. Kit [6] prior to confirmation test using GC-MS with head space method.
9000
Preparation of toluene standard
2000
13.392
8000 7000 6000 5000 4000 3000
1000
1 mL of toluene 99.7% was pipetted into a 100 mL volumetric flask and then made up to the mark with dichloromethane 99.7%. Toluene standard was sonicated in ultrasonic water bath.
0
5.00
10.00
15.00
20.00
25.00
30.00
35.00
Time-->
Retention Time (min)
Figure 1: GC-MS Head Space chromatogram for sample Urine 1.
Sample preparation
Results
Abundance
Abundance
Abundance
The urine samples were added with sodium fluoride as preservative were stored at 0-5°C. 10 mL of urine sample was placed in a head space bottle. The samples were heated in the oven at 100ºC for ½ hour. The headspace was withdrawn with a headspace syringe and injected at the GC-MSD.
TIC: SAMPLE 11.D\data.ms 3.292 20000 19000
17000
13.393
16000 15000 14000 13000
Local glue sample results
18.147
12000 11000
The percentage of abundance of toluene found in all 4 glue samples using Gas Chromatography Mass Spectroscopy (GC-MSD) using head space method are tabulated in Table 1 below.
10000
According to the results shown, toluene was the major compound that existed in all 4 glue samples analyzed.
5000
Toluene metabolite results
5.663
9000 8000 7000
20.118
6000
4000 3000 2000 1000 0
GC-MSD with headspace method was used for qualitative analysis for determination of toluene metabolites in this research. All the samples were analyzed by this method and the results are showed in Figure 1, 2, 3 and 4. The following figures 5 and 6 are GC-MS Head Space library chromatogram of N-(methylbenzoyl)-methyl esterglycine and p-cresol respectively. Glue brand
Substance detected
Abundance (%)
Dunlop
Toluene
91.1
GP
Toluene
91.1
Good Mark CA
Toluene
92.1
Wessbond
Toluene
91.1
Table 1: Substance detected and the abundance of the detected substance in glue samples using Gas Chromatography Mass Spectroscopy (GC-MSD) using head space method.
J Drug Metab Toxicol ISSN: 2157-7609 JDMT, an open access journal
15.922
18000
5.00
10.00
15.00
20.00
25.00
30.00
35.00
Time-->
Retention Time (min)
Figure 2: GC-MS Head Space chromatogram for sample Urine 2.
Discussion Local glue analysis The head space method easily separated the toluene substance compared to the usual Gas Chromatography Mass Spectroscopy Detector. This was because toluene has lower boiling point thus it easily evaporates making it a volatile compound that was suitable for head space method.
Urine samples analysis From the results (refer Figure 1-6), positive urine samples were 1, 2, 3 and 4. All of these positive samples contained p-cresol compound and urine samples 1 and 2 had N-(methylbenzoyl)-methyl ester-glycine. In
Volume 2 • Issue 3 • 1000112
Citation: Yacob AR, Zainalibdin MR, Said N (2011) Detection of Vapour Metabolites of Glue Sniffer’s Urine Using Head Space Gas Chromatography Mass Spectrometry. J Drug Metab Toxicol 2:112. doi:10.4172/2157-7609.1000112
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Abundance TIC: SAMPLE 15.D\data.ms 21000
107
100
20000 15.923
19000 18000 17000 16000
Abundance
15000
18.147
14000 13000
OH 50
12000 11000 10000 9000
77
8000
79
7000 6000
80
5000 4000
0
78
81
90 85
70 80 (replib) Phenol, 4-methyl-
3000 2000 1000 0
74 76
5.00
10.00
15.00
20.00
25.00
30.00
35.00
Time-->
105
92 90
100
110
120
Figure 6: GC-MS Head Space library chromatogram of p-cresol (ref. NISTO8 database).
Retention Time (min)
Figure 3: GC-MS Head Space chromatogram for sample Urine 3. Primary metabolism pathway
Abundance TIC: SAMPLE 16.D\data.ms 21000 20000 19000 18000 17000 16000
~ 1%
15.923
Abundance
5.661 15000 3.286 6 14000 13000 12000 11000 10000 9000
18.148
N-(methylbenzoyl)-methyl ester-glycine
13.393
8000 7000
-cresol
6000 5000
Minor metabolism pathway
4000 3000
Figure 7: Reaction step for toluene metabolism.
2000 1000 0
5.00
10.00
15.00
20.00
25.00
30.00
35.00
Time-->
Retention Time (min)
Figure 4: GC-MS Head Space chromatogram for sample Urine 4.
119
100
O NH
50
O
The following reactions were deduced based on the results obtained from the study. Figure 7 show the most probable reaction step of metabolism of toluene based on the results.
O 91 0
207
148 105
131
160
175 187
100 120 140 160 180 (replib) Glycine, N-(4-methylbenzoyl)-, methyl ester
200
220
Figure 5: GC-MS Head Space library chromatogram of N-(methylbenzoyl)methyl ester-glycine (ref. NISTO8 database).
J Drug Metab Toxicol ISSN: 2157-7609 JDMT, an open access journal
this case, the N-(methylbenzoyl)-methyl ester-glycine has a similar structure as a hippuric acid. This proves that urine samples 1 and 2 have a high concentration of hippuric acid. The p-cresol compound present in urine samples 1, 2, 3 and 4 is one of toluene metabolites. Approximations of 0.4-1.1 % p- and o-cresol are produced when an individual is exposed to toluene [8]. Thus, the detection of p- cresol in this amount range may indicate that the person was exposed to high concentration of toluene or had sniffed toluene vapour directly. As the results indicated, it was proven that the samples 1, 2, 3 and 4 have been exposed to toluene and indicated that these persons were glue sniffers.
Conclusion The results show that toluene is the major solvent found in glue samples analyzed. Toluene is used in industry for the synthesis of the
Volume 2 • Issue 3 • 1000112
Citation: Yacob AR, Zainalibdin MR, Said N (2011) Detection of Vapour Metabolites of Glue Sniffer’s Urine Using Head Space Gas Chromatography Mass Spectrometry. J Drug Metab Toxicol 2:112. doi:10.4172/2157-7609.1000112
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chemical compound and as the solvent for paint, print ink and adhesive. In the glue samples, toluene is indicated as the solvent for adhesive materials that are included in this group. In this case, the research objectives are the determination and analysis of the major component of glues, which is toluene. Because of that the study will focus only on toluene sample and its derivatives that may be present in urine samples after sniffing the glue. Toluene metabolites such as p-cresol and N-(methylbenzoyl)-methyl ester-glycine were demonstrated to be present in urine samples of individuals that have been glue sniffing prior to their urine being sampled. GC-MS with head space method has proven to be an easier confirmation test in comparison to GC-MSD as the analytic are volatile compounds. Acknowledgement Special thanks to National Anti Drugs Agency (AADK) for their willingness to collaborate in this research. This study was made possible with the financial support from Ministry of Science Technology and Innovation (MOSTI) and Department of Chemistry, Johor branch.
References 1. Nicholas K, Zili Sloboda, Maria De La Rosa (1995) Epidemiology of Inhalant Abuse: An International Perspective Journal of Inhalant abuse 2-3. 2. Navaratnam V. (1988) The Problem of Inhalant Abuse in the South-East Asian/ Western Pacific Region- A Regional Analysis. 16-23. 3. Yacob AR, Zainalibdin MR (2009) Characterization using instrumentation of local glue related glue sniffing activity. Indian Journal of Forensic Medicine and Toxicology 3(2): 58-64. 4. Arif AE, Navaratnam V (1988) Abuse of Volatile Solvents and Inhalants. Drug Research Centre 16-23. 5. Sharifah MTF (1990) Report on the Inhalant Abuse Problem in Sarawak, East Malaysia. 12-16. 6. Yacob AR, Zainalibdin MR (2010) G.S. Kit: Mobile test kit for glue sniffing screening. Advanced Management Science 399-402. 7. Saad S, Pakiam C (2005) Screening for Volatile Solvents in Blood by Headspace Gas Chromatography. JKM F0540: 1-5. 8. Yamazaki K, Tanaka E, Misawa S (1992) Urinary ortho-cresol Concentrations as an Indicator of Toluene Inhalation in Glue Sniffer. JFor.Sci 32: 215-223.
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