![Microcrystal tests
Procedures have been reported using mercury chloride [33] and mephedrone was
observed to form characteristic “paddlewheels and rosettes of blades”
Method
An aliquot (10 μL) of the test solution (1 g/L) is
mixed with 10 μL of the reagent
on a glass slide. A plastic pipette is used to aid
nucleation and crystal formation.](https://image.slidesharecdn.com/untitledpresentation-200506182857/85/DETECTION-METHODS-FOR-SYNTHETIC-CANNABINOIDS-AND-CATHINONES-13-320.jpg)
DETECTION METHODS FOR SYNTHETIC CANNABINOIDS AND CATHINONES
- 1. DETECTION METHODS OF SYNTHETIC CANNABINOIDS AND CATHINONES SAFA RUKSHAR
- 2. SYNTHETIC CANNABINOIDS Synthetic Cannabinoids are among recently synthetized medicines. Cannabis are most illegally use for all over world for many years. These Synthetic Cannabinoids complexes were used by scientists to isolate the medicinal effects of natural cannabis. Synthetic cannabinoids are chemicals that bind to cannabinoid receptors and produce effects like to those of tetrahydrocannabinol.
- 3. Synthetic Cannabinoids are mentioned to as materials with mechanical feature which permit required to one of the recognized cannabinoid receptors, i.e. CB1 or CB2, present-day in human cells and complexes with similar chemical structures. The CB1 receptor is placed mostly in the brain and spinal cord and is accountable for the representative physiological and mainly the psychotropic belongings of cannabis, whereas the CB2 receptor is located mainly in the spleen and cells of the immune System and may mediate immune‐modulatory effects.
- 4. COLORIMETRIC TESTS The Marquis reagent, which reacts with all nitrogen-containing drugs, is positive. Dragendorff reagent is also positive . Fast blue BB reacts with cyclohexylphenols, and the LOD concentration is not lower than that of Marquis reagent. Although it is possible to detect synthetic cannabinoids with each reagent in these screening tests, it is difficult to detect small amounts or mixtures of synthetic cannabinoids
- 5. Immunochemical detection ELISAs developed in-house could be calibrated at 5 ng/ml with the 5-OH and 4- OH metabolites respectively, and evaluated for the detection of synthetic cannabinoids in urine. Recently, some commercially available immunoassay kits, such as DrugCheck K2/Spice Test, DrugSmart Cassette, and RapiCard InstaTest, have been developed for the detection of these drugs in urine. These devices are more useful than the colorimetric methods, because they do not require special reagents or tools, and the results are obtained easily
- 6. GC–MS detection Molecular and/or fragment ions observed by full scan data acquisition of GC–MS reflect the structures of the synthetic cannabinoids. The identification of synthetic cannabinoids is facilitated by comparison of the spectra with commercial and open databases.
- 7. LC–MS-MS detection Many research groups have used LC–MS-MS for determination of synthetic cannabinoids in herbs and biological samples, and some have studied the fragmentation of synthetic cannabinoids in detail. Because the protonated molecular ion is only observed by LC–MS, and the information acquired by LC– MS is lesser than that for GC–MS, it is necessary to obtain other data that reflect the chemical structures by LC–MS-MS.
- 8. CATHINONES Cathinones, can be isolated from the khat plant or produced by synthetic means. Cathinone analogs with high selectivity and strong activity for serotonin receptors and monoamine transporters have been distributed in the drug market. Cathinones normally present as white or off-white powders although they can come in a range of colours. Mephedrone, for example, commonly appears as white or yellow powder/crystals, with a distinct odour described as ranging from fishy to vanilla or bleach. Although primarily encountered as a powder, mephedrone has also been known to take the form of capsules/tablets of varying design. Mephedrone (4-MMC), M-CAT), Methylone, Methcathinone, Buphedrone, Bupropion, Pyrovalerone, Alpha-Pyrrolidinovalerophenone (alpha-PVP).
- 9. Extraction and sample preparation Samples should be prepared according to their morphology as follows: Powders: A solution should be prepared at a concentration of approximately 1mg/ mL in methanol. Tablets: A representative number of tablets (following the sampling procedure) should be ground to a fine powder and a solution prepared as for powders. Capsules: The contents of a representative sample of capsules (following the sampling procedure) should be removed and a solution prepared as for powders. Syringes or glassware: Should be washed with a minimum amount of methanol.
- 10. TEST FOR SYNTHETIC CANNABINOIDS Various test used for traditional or main components of cannabis test negative for synthetic cannaboids, The use of 2,4-dinitrophenylhydrazine, which reacts with a keto moiety, is capable of reacting with synthetic cannabimimetics, such as the naphthoylindole, phenylacetylindole, benzoylindole, and cyclopropylindole classes, either in powder form or adsorbed onto plant material, and a positive test solution turns from yellow to orange. Marquis reagent gives positive h Dragendorff reagent is also positive Fast blue BB reacts with cyclohexylphenols, ELISAs developed in-house could be calibrated and evaluated for the detection of synthetic cannabinoids in urine.
- 11. TEST FOR CANTHINONES Presumptive tests are non-specific tests that can be used to identify which class of compounds a substance belongs to. However, they cannot be used to identify a specific compound within that class. Therefore, confirmatory tests must always be carried out in conjunction with these preliminary tests.
- 12. Zimmermann test reagents Different cathinones gives different color when coming in contact with the reagent. Color change shows a positive test for cathinones. For intance, Mephedrone gives Dark red/purple Benzedrone (4-MBC) gives pale pink.
- 13. Microcrystal tests Procedures have been reported using mercury chloride [33] and mephedrone was observed to form characteristic “paddlewheels and rosettes of blades” Method An aliquot (10 μL) of the test solution (1 g/L) is mixed with 10 μL of the reagent on a glass slide. A plastic pipette is used to aid nucleation and crystal formation.
- 14. Thin layer chromatography (TLC) Solvet system - Ethyl acetate, methanol and (25%) ammonia – (85:10:5 v/v/v). Standards should be prepared at a concentration of between 1 to 5 mg/mL in methanol. Visualization The plate should be viewed under UV light (254 nm) with any spots being noted before being sprayed with ninhydrin reagent (2%).
- 15. Gas chromatography (GC) with mass spectrometry (MS) Preparation of the internal standard solution Eicosane (or a similar n-alkane) can be used as an internal standard and prepared as a solution in methanol at a concentration of 1 mg/mL. GC-MS operating conditions GC oven conditions: 90°C for 1 minute, increased to 300°C at a rate of 8°C/min. and then held isothermal at 300°C for 10 minutes Column: 5% phenyl / 95% methyl silicone column (HP-5MS), 30 m length x 0.25 mm i.d., 0.25 μm film thickness Injection parameters: 2 μL aliquot of sample injected with a split ratio of 75:1 Injector temp.: 225°C Carrier gas: Helium, flow rate: 1.0 mL/min. MS source temp.: 230°C
- 16. High performance liquid chromatography Column: HiChrom ACE 3 C-18, 150 x 4.6 mm i.d., 3 μm particle size Isothermal at 22°C Mobile phase: 28:72 (v/v) methanol: 10 mM ammonium formate (adjusted to pH = 3.5 with formic acid) Flow rate: 0.8 mL/min. Detection: Photodiode array-UV detector (258 nm for cathinones) Injection volume: 10 μL Internal standard: Nicotinamide, 2.5 μg/mL
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