Analysis of powder samples for presence of psychoactive ingredients – Screening report
Section 1: Investigative Report
Section 2:Identification, confirmation and quantification
Identification
RRT [CALCULATE RELATIVE RETENTIONTIME}
Kovats Indices (KI)
Oven temperature program of the Gas Chromatography:
150°C to 205°C at 5° C per minute (first 11 minutes), hold for 6 minutes, ramp to 280°Cat 25°C per minute (last 3 minutes). Total run time 20 minutes.
The temperature program was non isothermal hence the PTGC KI equation was used.

GC-FID Standard Drugs + n-alkane ladder
Substance Retention time (minutes)
C14 3.014
Benzocaine 4.541
C16 5.042
Phenacetin 6.505
C18 7.998
MDA 8.259
Caffeine and Lidocaine 9.197
C20 11.497
Methadone 14.664
C22 16.523
C24 19.799

GC-NPD Standard drugs + n-alkane
Substance Retention time (minute)
Ephedrine 4.078
Benzocaine 6.428
MAD I 6.759
Phenacetin 8.364
Caffeine 11.021
Lidocaine 11.772
Methadone 18.468

Sample 1 GC-FID results
Substance Retention time (minute)
C12 2.007
C14 3.093
C16 5.118
C18 8.071
Unknown peak a 9.283
C20 11.544
Unknown peak b 14.698
C22 16.571
C24 19.803
Unknown peak a: Identified as Caffeine and Lidocaine
Unknown peak b: Identified as Methadone
Sample 1 NPD results
Substance Retention time (minute)
Unknown peak a 11.043
Unknown peak b 11.787
Unknown peak c 18.477
Unknown peak a: Identified as caffeine
Unknown peak b: Identified as Lidocaine
Unknown peak c: Identified as Methadone
KI for sample 1 using FID results of unknown peak b

Sample 2 GC-FID results
Substance Retention time (minute)
C12 1.981
C14 3.036
C16 5.034
C18 7.989
Unknown peak a 9.163
C20 11.478
Unknown peak b 14.634
C22 16.484
C24 19.789
Unknown peak a: Identified as Caffeine and Lidocaine
Unknown peak b: Identified as Methadone
Sample 2 NPD results
Substance Retention time (minute)
Unknown peak a 10.996
Unknown peak b 11.742
Unknown peak c 18.457
Unknown peak a: Identified as caffeine
Unknown peak b: Identified as Lidocaine
Unknown peak c: Identified as Methadone
Kovats (KI) for sample 1
tr(u) = 14.698 mins
tr(x) = 11.544 mins
tr(x+n) = 19.803 mins
n = 4 (24-20)
x = 20
KI = 100 [4 (14.698-11.544) / (19.803-11.544) +20]
KI = 100[4 (3.154/8.259) +20]
KI = 100[4 (0.3818864269) +20]
KI = 100 [21.52754571]
KI = 2152.755 mins
KI for sample 2 = 2151.895 mins
In order for KI to be accepted it must be within ±1%
[WHAT IS THE KI OF METHADONE ACCORDING TO CLARKE’S] reference this
Confirmation of Methadone
Fast-Gas Chromatography – Mass Spectrometry Results
Three peaks were identified and their retention time measured. The peak retention times were compared using the AMDIS chromatogram database search.
Peak Retention time minutes Peak ID based on library database
2.97 Caffeine
3.04 Lidocaine
3.58 Methadone

Retention time (RT) of the Methadone reference standard was 3.59. The RT of the sample was 3.58 mins which is within the 1% range of the standard. This identifies and confirms methadone.
In order to further confirm methadone, the confirmation ratio PA m/z 72 ÷ PA m/z 294 was used.
Methadone PA m/z 72 PA m/z 294
Sample 15076347 81011
Reference Standard 9018647 44267
In order to work out the tolerance window, the relative abundance of methadone reference standard of m/z 294 relative to m/z 72 was calculated.
Assuming PA m/z 72 = 100%
PA m/z 294 = x
X = (PA m/z 294 × 100) ÷ PA m/z 72
X = 44267 × 100 ÷ 9018647
X = 0.491%
This is less than 50% hence according to the table in SOP/UG/DoA/08 the tolerance is within ±10%.
Confirmation ration of reference standard = 9018647 ÷ 44267
= 203.733
± 10% of 203.73 puts the range between 183.3507 to 224.1063
Confirmation ratio of sample = 186.102
This falls within the tolerance window hence the results can be accepted.
QUANTIFICATION
Quantification ratio = PA m/z 72 / PA m/z 78
Standar (μg/mL) PA m/z 72 PA m/z 78 PA m/z 294 PA m/z 303 Quantification ratio
0 0 3088545 0 31752 0
50 1206959 2947267 13566 30075 0.409518038
100 2651085 3617143 30662 31232 0.732922364
500 4242834 1853366 52341 19523 2.289258571
Pos. Ctl. (300 μg/mL) 2491817 1699803 30251 17685 1.465944583
Sample 1 3410035 2753322 40308 27994 1.238516599
Sample 2 2056028 2435611 23844 24929 0.844152863

Equation of the line : y = 0.0043x + 0.1528
Therefore x = y – 0.1528 ÷ 0.0043
Where y = peak are ratio and x = methadone concentration μg/mL.
Concentration of positive control = 303.208 μg/mL
Concentration of sample 1 = 252.492 μg/mL
Concentration of sample 2 = 160.780 μg/mL
% w/w
Sample 1
Calculated concentration 252.492 μg/mL
However in step 5 of SOP/UG/DoA/09 the sample was diluted by 10
Therefore the actual concentration is 252.492 μg/mL × 10
= 2524.92 μg/mL
Convert to mg/mL = 2.52492 mg in 1mL
The extract was done in 1.5mL
Concentration in 1.5mL
= (2.52492 mg × 1.5mL) ÷ 1mL
= 3.787mg in 1.5mL
This mass comes from the original 15mg measured in SOP/UG/DoA/04
15mg represents 100% weight by weight
Hence 3.787mg represents 25.25%
Sample 2 %w/w = 16.08%

Section 3: Item receipt and initial examination
On receiving the item, SOP/UG/DoA/01 was followed. The tamper evident packaging and description of the item was checked in order to confirm consistent chain of custody. The item was then signed for, a chain of custody information completed and item entered in the laboratory database for received items.The case no: D/1020/16 and serial no: DOA15/15/16/PI were also recorded. Sample was a white powder in a plastic bag, and appeared to be homogenous. Weight of the sample including the container was 0.8820g. The sample was then analysed using a light microscope at 4× magnification and 10× magnification, under the microscope the samples appeared as black solid crystals. Once an item of physical evidence has been removed from the location at which it was found, it must be packaged in such a way that it cannot become contaminated or damaged. It is equally important that the item is unable to escape from its packaging for two reasons. First, to preserve the item from contamination in an unprotected environment. Secondly, to prevent the item contaminating other packaged evidence. For example, imagine the consequences of a suspected drug in the form of a powder escaping from its package. This could end up (and no doubt will end up) on the outside of another package so that when this is handled, that drug becomes transfered to the person who opens the package and then on to other items for examination.
It was necessary to record information such as the serial number in order to ensure that the item can be tracked. The sample weight was important as well as measuring the amount used at each stage of analysis. This will ensure that if any amount were to be taken from the sample without it being documented it would hopefully be detected (hence preventing theft of the sample).
The container containing the drug sample was a plastic bag. Further examination of the container include dusting for finger-marks, DNA from hair, body fluid or skin cells and ore presence of fibres. Results from further analysis of the packaging can provide information which could link the drugs to people, a place and other batches of seized drugs.
Plastic packaging materials (e.g. bags and cling film) are commonly used to wrap quantities of illicit drugs, for storage or to sell as ‘deals’. When those plastic packaging materials are produced the manufacturing process will leave ‘marks’ on that packaging, which may be characteristic of a particular piece of manufacturing equipment and a particular time period in one manufacturing run, on that equipment. Those manufacturing ‘marks’, along with other physical and chemical characteristics of the packaging material can be used to compare samples of packaging and possibly link them evidentially to a common source[REFERENCE THIS}

FUTHER QUESTIONS TO BE ANSWERED
Section 4 – FTIR (10 marks)
FTIR was used as a downgraded technique for screening. Devise an analytical strategy to use IR as a category A technique for confirmation of the substances you found in the sample (methadone, lidocaine, caffeine and sodium carbonate).
Support your answer with at least 3 examples from the scientific literature (Attach the related reference section directly below this section; References do not contribute to the word limit).
Word limit: 300.
Section 5 – HPTLC (10 marks)
HPTLC was followed by a colour development obtained by spraying the plate with a reagent (iodoplatinate reagent).
Using the scientific literature, find one alternative analytical process which could be applied to the TLC plate to further characterise the sample. Describe this technique, and briefly explain its usefulness for drug analysis. (Attach the related reference section directly below this section; References do not contribute to the word limit).
Word limit: 300.
Section 6 – GC-NPD (10 marks)
(1) Propose a ladder useable for NPD for its application to the calculation of Kovats indices.
(2) Briefly explain why, for the same analyte, the RTs were different between the FID and the NPD chromatograms.
(3) Briefly explain why the peak shape of certain substances was different for the channel with the FID and the channel with the NPD.
(4) Briefly explain the value of an NPD when analysing a fingerprint suspected of amphetamine contamination.
Word limit: 300.
Section 7 – GC-MS quantification (15 marks)
(1) There were no acceptance of results criteria for the quantification of methadone on the SOP you followed (ULFCL – SOP/UG/DoA/09), explain what you used to validate your results. Show all working out.
(2) Suggest how you would adapt ULFCL-SOP/UG/DoA/09 for the quantification of lidocaine and caffeine. Specify and justify all technical details.
Word limit: 300.