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Introduction: ILLICIT Drug Identification: HPLC Analysis
In this chapter, the lab report is based on the identification of illicit drugs that are used by males aged 18 and 22. To identify the powder the laboratory test is important. The objectives of the study are discussed here. The main objectives are-
To identify the illicit drug, the lab technician will have to use “high-performance liquid chromatography” or HPLC. The identification of the drugs should primarily be based on “retention time,” which is calculated in the laboratory.
To create a standard calibration set. This is important for the calculation of the amount of the drug. The drug will have to be used as a diluted sample. From the calculated and observed graph, the “peak areas” of the graphs will have to be observed, and based on these peak areas the quantification of the drug should have to do. The % purity of the samples will have to be calculated.
To report the analytical data that is collected in the laboratory. The data have to be collected based on ATR-FTIR spectroscopy in the laboratory. The reflectance of the FTIR spectroscopy will have to be calculated.
Materials and methods
Materials or reagents that are used in this experiment
Reagents
Amount of reagents
Cocaine standard
1 mg/mL
Ketamine standard
1 mg/mL
Benzocaine standard
1 mg/mL
Unknown sample’s extract
1 mg/mL
In the above chart, the reagents that are used in the laboratory for this experiment are discussed. The above-mentioned reagents will have to be used in the mobile phase that is used in the HPLC experiment of chromatography.
Methods that are used in the experiment
The syringe of injection in which the diluted sample will have to take for the experiment will have to be rinsed first, with the help of deionized water. After rinsing the sample will have to be injected into the instrument of HPLC.
The above-mentioned step will have to carry forward for each of the three unknown samples. In this case, the three samples are cocaine, benzocaine, and ketamine. The lab technician will have to be identified the samples between these three samples. The instrument that is important for the experiment will have to be ready before the injection of the unknown samples.
The chromatograms of the samples will have to be collected. After the collection process is done then the lab technician will have to make notes about the column type, HPLC manufacturer, the composition of the mobile phase, and the wavelength of the detector will also have to be calculated.
After the above steps, the analysis of the chromatograms will have to be done. Then the components can be identified with the help of a chromatogram.
Then the illicit drug will have to be identified.
A standard calibration set will have to be produced.
Then a calibration curve will have to be determined.
Concentration (microgram/mL)
The volume of stock of solution (mL)
The volume of deionized water (mL)
Final volume (mL)
0
0
1
1
25
0.25
0.75
1
50
0.50
0.50
1
75
0.75
0.25
1
100
1
0
1
Table 1: A standard calibration table
Result and discussion
Figure 1: HPLC chromatogram for an unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 4.968 min. The area of the highest peak is 6240.44 mAU and the height of the highest peak is 469.13nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.578
156.65
37.04
2.37
2.787
120.45
26.33
1.82
3.399
41.87
8.80
0.63
4.968
6240.44
469.13
94.31
5.771
19.78
2.39
0.30
9.436
37.94
3.45
0.57
Table 2: Report of the chromatogram
Figure 2: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 4.916 min. The area of the highest peak is 8232.53 mAU and the height of the highest peak is 557.55nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.573
221.47
48.25
2.54
2.780
134.91
27.56
1.55
3.392
47.67
10.49
0.55
4.916
8232.53
557.55
94.43
5.760
28.46
3.22
0.33
9.433
53.08
4.21
0.61
Table 3: Report of the chromatogram
Figure 3: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 5.013 min. The area of the highest peak is 4315.04 mAU and the height of the highest peak is 375.20nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.216
24.54
2.63
0.53
2.585
135.50
32.49
2.94
2.792
81.26
17.81
1.76
3.406
36.36
7.41
0.79
5.013
4315.04
375.20
93.51
5.780
22.05
2.52
0.48
Table 4: Report of the chromatogram
Figure 4: HPLC chromatogram for an unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 4.953 min. The area of the highest peak is 6297.40 mAU and the height of the highest peak is 474.92nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.580
210.05
46.85
3.13
2.786
116.24
25.83
1.73
3.398
57.31
11.38
0.85
4.953
6297.40
474.92
93.81
9.412
32.15
2.85
0.48
Table 5: Report of the chromatogram
Figure 5: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 3.714 min. The area of the highest peak is 24.76 mAU and the height of the highest peak is 1.48nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
3.714
24.76
1.48
100.00
Table 6: Report of the chromatogram
Figure 6: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 5.011 min. The area of the highest peak is 4448.70 mAU and the height of the highest peak is 381.15 nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.211
30.81
3.96
0.65
2.585
137.28
33.04
2.88
2.793
83.72
17.61
1.76
3.407
40.64
7.63
0.85
5.011
4448.70
381.15
93.29
9.420
27.29
2.46
0.57
Table 7: Report of the chromatogram
Figure 7: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 2.133 min. The area of the highest peak is 14858.35 mAU and the height of the highest peak is 2870.16 nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area%
2.133
14858.35
2870.16
99.24
5.235
49.23
4.81
0.33
6.832
22.18
1.34
0.15
7.971
19.23
1.32
0.13
9.550
23.51
1.47
0.16
Table 8: Report of the chromatogram
Figure 8: HPLC chromatogram for unknown sample
From the chromatogram it is observed that the retention time for the highest peak is 2.211 min. The area of the highest peak is 21917.63 mAU and the height of the highest peak is 3737.87 nm. The wavelength used in the chromatogram is 217 nm.
RT [min]
Area
Height
Area %
2.211
21917.63
3737.87
100.00
Table 9: Report of the chromatogram
Conclusion
After the analysis of the above data it is clear that the time of retention for cocaine peaks at 4.916 min. The retention time for ketamine is 2.133, and for benzocaine is 2.211 min. The time of retention for the unknown sample is 4.968 min. The wavelength used in the chromatogram is 217 nm. The reflectance of the FTIR spectroscopy will have to be calculated.
References
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DG Watson, 2020 Pharmeceutical Analysis Available at http://repository.UKbooks.id/id/eprint/275/1/MCU_2017_Pharmaceutical_Analysis_A_Textbook_for_Pharmacy_Students.pdf [Accessed on 12.04.2023]
Journal
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Zhu, L., Zhao, Y., Liu, T., Chen, M., Qian, W.P., Jiang, B., Barwick, B.G., Zhang, L., Styblo, T.M., Li, X. and Yang, L., 2022. Inhibition of NADPH oxidase-ros signal using hyaluronic acid nanoparticles for overcoming radioresistance in cancer therapy.ACS nano,16(11), pp.18708-18728.
Article
Orsolini, L., Corkery, J.M., Chiappini, S., Guirguis, A., Vento, A., De Berardis, D., Papanti, D. and Schifano, F., 2020. ‘New/designer benzodiazepines’: an analysis of the literature and psychonauts’ trip reports.Current neuropharmacology,18(9), pp.809-837. Accessed from https://www.unodc.org/documents/scientific/STNAR47_Piperazines_Ebook.pdf [Accessed on 12.04.2023]
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