Friday, June 8, 2007

Lab Analysis of Marijuana

The specificity of a marijuana analysis is still a widely discussed topic among those in the forensic and legal communities. In the course of the past 25 years, the concensus of opinion concerning the analysis of marijuana has remained fairly consistent. In those situations where plant material is encountered, the marijuana is first examined using a stereomicroscope. The presence of the bear claw cystolithic hairs and other histological features are noted using a compound microscope.

The plant material is then examined chemically using Duquenois – Levine reagent in a modified Duenois Levine testing sequence. These two tests are considered to be conclusive within the realm of existing scientific certainty in establishing the presence of marijauana.3–5. The Modified Duquenois–Levine test is conducted using Duquenois reagent, concentrated hydrochloric acid, and chloroform. The Duquenois reagent is prepared by dissolving 2g of vanillin and 0.3 ml of acetaldehyde in 100 ml of ethanol. Small amounts (25 to 60 mg is usually sufficient) of suspected marijuana leaf is placed in a test tube and approximately 2 ml of Duquenois reagent is added. After 1 min, approximately 1 ml of concentrated hydrochloric acid is added. Small bubbles rise from the leaves in the liquid. These are carbon dioxide bubbles produced by the reaction of the hydrochloric acid with the calcium carbonate at the base of the cystolithic hair of the marijuana. A blue to blue-purple color forms very quickly in the solution.

Approximately 1 ml of chloroform is then added to the Duquenois reagent/ hydrochloric acid mixture. Because chloroform is not miscible with water, and because it is heavier than water, two liquid layers are visible in the tube—the Duquenois reagent/hydrochloric acid layer is on top, and the chloroform layer is on the bottom. After mixing with a vortex stirrer and on settling, the two layers are again clearly distinguishable. However, the chloroform layer has changed from clear to the blue to blue-purple color of the Duquenois reagent/hydrochloric acid mixture.

One variation in this testing process involve pouring off the Duquenois reagent sitting in the tube with the leaves before adding the hydrochloric acid. The remainder of the test is conducted using only the liquid. Another variation involves conducting the test in a porcelain spot plate. This works, although some analysts find the color change a bit more difficult to detect. A third variation involves extracting the cannabis resin with ether or some other solvent, separating the solvent from the leaves, allowing the solvent to evaporate, and conducting the Modified Duquenois–Levine test on the extract.

Marquis reagent is prepared by mixing 1 ml of formaldehyde solution with 9 ml of sulfuric acid. The test is done by placing a small amount of sample (1 to 5 mg) into the depression of a spot plate, adding one or two drops of reagent, and observing the color produced. This color will usually be indicative of the class of compounds, and the first color is usually the most important. A weak reponse may fade, and samples containing sugar will char on standing because of the sulfuric acid. Marquis reagent produces the following results:

1. Purple with opiates (heroin, codeine).
2. Orange turning to brown with amphetamine and methamphetamine.
3. Black with a dark purple halo with 3,4-methylenedioxyamphetamine (MDA) and
3,4- methylenedioxymethamphetamine (MDMA).
4. Pink with aspirin.
5. Yellow with diphenhydramine.

A thin-layer chromatographic (TLC) analysis, which detects a systematic pattern of colored bands, can then be employed as an additional test.6,7 Though it is not required, some analysts will run a gas chromatograph/mass spectrometrometer (GC/MS) analysis to identify the cannabinoids in the sample.

The solvent insoluble residue of hashish should be examined with the compound microscope. Cystolythic hairs, resin glands, and surface debris should be present. However, if most of the residue is composed of green leaf fragments, the material is pulverized marijuana or imitation hashish.

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