Substance Abuse PHG 414 - PSAU · Hallucinogens 1-Peyote cactus Mescaline Tolerance : It builds...

Prof. Dr. Amani S. Awaad

Professor of Pharmacognosy

Pharmacognosy Department,

College of Pharmacy Salman Bin Abdulaziz University,

Al-Kharj. KSA.

Email: [email protected]

Substance Abuse

PHG 414

Welcome Back

Dears

Hallucinogens

You should be able to know:

• What is Peyote cactus ,contents, effect and analysis

• Identify contents, effect and an of Peyote cactus

• What is Psilocybe Mushroom, contents, effect and analysis

Catecholamine-Like Hallucinogens

Mescaline

1-Peyote cactus

Hallucinogens

-Lophophora williamsii, family cactaceae was important to the

ancient Aztecs as ritual hallucinogen.

- They call this cactus” peyotl” meaning “furry thing” a reference to

the tufts of hairy crowning the mature plants.

-Peyote use is now an important part of religious ceremonies of the

Native American church of North America causes concomitant

hallucinations and euphoria.

-The peyote occurs isolated or in groups usually on limestone soils, on

rocky slopes and dried river beds

Mescal buttons: (peyote buttons), the form most commonly found in

the illicit traffic, are the dried, brown, disk-shaped tops of the cactus.

1-Peyote cactus Peyote /Mescal Button/Lophophora williamsii

http://www.crazycacti.co.uk/LF002-Lophophora-Diffusa.html

Hallucinogens 1-Peyote cactus

Chemical contents:

Mescaline (3,4,5-trimethoxyphenyl ethylamine) the active chemical of

peyote, is a dark-brown powder ground from "buttons" of the Mexican

cactus peyote (Lophophora williamsii ).

It can also be created synthetically (from 3,4,5-trimethoxybenzaldehyde,

nitromethane, cyclohexylamine and acetic acid, initially) forming white

mescaline sulfate crystals.

O C H 3

O C H 3

O C H 3

N H 2

1

234

56

M e sca lin e

-UP TO 30% of the total peyote alkaloids are present as mescaline.

-It appears mostly as the SO4 or HCl salt from time on the illicit

drug markets in the USA and Europe.

-The minor peyote alkaloids are N-methyl mescaline, 3-O-

demethyl-mescaline and hordenine.

-About 30 additional alkaloids e.g. pellotine, anhalonidine and

lophopharine belong to the group of the structurally related

tetrahydro isoquinoline alkaloid.

-The minor alkaloids do not show any psychotropic activity N-methyl mescaline 3-O-demethyl-mescaline


Mescaline Tolerance : It builds with repeated usage, lasting for a few days. Mescaline

causes cross-tolerance with other serotonergic psychedelics such as LSD and psilocybin.

Chemical contents:

Mescaline excretion: About half the initial dosage is excreted after 6 hours, and between

20% and 50% of mescaline is excreted in the urine unchanged, and the rest being excreted as

the carboxylic acid form of mescaline. The LD50 of mescaline has been measured in various

animals: 212 mg/kg i.p. (mice), 132 mg/kg i.p. (rats), and 328 mg/kg i.p. (guinea pigs).

- Mescaline crystalline power can be dissolved into tea or placed in capsules.

- It causes hallucinogenic effects similar to LSD

Signs and Symptoms: Anxiety, hyperreflexia, static tremors, vivid visual hallucinations,

abdominal pain, nausea, and diarrhea.

Administration: Chewed raw or cooked and eaten OR Pure powder form

A dose of 350 to 500 milligrams produces hallucinations lasting from 5to 12 hours


Pharmacological Effect:

-The crown of peyote cactus is sliced into circular mescal buttons that are eaten.

-The mescaline is readily absorbed into the blood stream from GIT tract, but crosses the blood-

brain much more slowly.

-Therefore, relatively high dose of mescaline are necessary to produce hallucinogenic effects.

-The need for high doses and relative impermeability of the blood-brain barrier to mescaline

account for drugs long duration of activity (as long as 12 hours).

- Mescaline is eliminated primarily in the urine

-mescaline produces physiological and psychological responses similar to (LSD) .

-Physiological responses include the dilatation of the pupils of the eyes, the elevation of

temperature, increase heart rate, and increase blood pressure.

-Psychological effects include hallucinations in vivid colors of designs, flowers and animals.

-There are also distortions in the perception of colors, time and space.

-Some individuals experience serious nausea and vomiting.

- In spite of this, mescaline is relatively safe, an extremely large dose is needed to produce a

toxic response or cause death.


Mode of action:

Mescaline acts similarly to other psychedelic

agents. It binds to and activates the serotonin 5-HT2A

receptor with a high affinity as a partial agonist.

How activating the 5-HT2A receptor leads to psychedelia is

still unknown, but it likely somehow involves excitation of

neurons in the prefrontal cortex. Mescaline is also known to

bind to and activate the serotonin 5-HT2C receptor.

In addition to serotonin receptor activity,

mescaline also stimulates the dopamine receptors.

Whether mescaline possesses dopamine receptor agonist

properties or initiates the release of dopamine remains

unclear.


Identification and Analysis

There are different methods for analysis of Peyote cactus which can be as following:

a. Morphological examination b. Chemical examination c. Chromatographic examination.

a. Macro and Microscopic characters:

-Peyote is a simple spineless grayish-green cactus.

- It attains 2-7 cm height and 4-12 cm in diameter. The roots are usually 8-11 cm long.

- The crowns are spineless, globular, top-shaped or sometimes flattened 2-8 cm in diameter with 8-10

well defined ribs.

-The pale pink, rarely whitish flowers 1.5-2.5 cm in diameter, are located at the outer of the crown.

-Powder cactus does not show enough characteristics and is therefore in appropriate for forensic

purposes.

b. Chemical examination

Color tests:-Positive color tests are not indications on the presence of mescaline, other

material like phenylethylamines, amphetamines as well as other non harmful materials

can give positive test.

Marquis’ Test: solution-1: mixture of 8-10 drops of formaldehyde and 10 ml glacial acetic

acid. Solution-2 : concentrated H2SO4

Place to small amount of the suspected material (from powder, tablet, capsule) placed in a spot

plate, add 1 drop of HCHO solution and 2 drops of H2SO4 . an orange-red color indicates the

possible presence of mescaline. the detection limit is about 10 µg.


Identification and Analysis c. Chromatographic examination

For TLC it is not necessary to perform a further clean up of the extracts if

ninhydrine is used as spray reagent, filter and concentrate with Et2O.

• For mescaline powder: both free base and salts (SO4 and HCl) are soluble in MeOH and

this is the solvent of choice for sample preparation for qualitative and quantitative analysis.

• For mescaline tablets and capsules, representative sample of the finely powdered tablets or

the contents of capsules is extracted by shaking or sonicating with MeOH to obtain after

filtration a solution of about 1mg/ml.

TLC: the prepared methanolic extract is spotted on 2 silica gel GF254 plates alongside a standard

solution of mescaline. Solvent system: CHCl3:MeOH:NH4OH (82:17:1) MeOH:NH4OH (100:1.5)

Develop and dry TLC, visualize first by UV light at 254 nm followed by spray by either

ninhydrine or fluorescumine. When using fluorescamine, the plate has to be dried after spraying

then observe the plate at 365 nm it will give bright yellow fluorescent spot intensify by NH3.

•When using ninhydrine, the plate has to be heated after spraying in an oven at 120°C for 15

minutes it will give violet.

Extraction Procedures: For peyote cactus and mescal buttons, mescaline is quantitively extracted from dried and pulverized

plant material by shaking or sonication with MeOH: NH4OH (99:1).


Identification and Analysis

c. Chromatographic examination

GLC: column fused silica capillary column, chemically bonded and cross-linked

methylphenyl silicone.

Carrier gas He, detector FID. Quantitation by peak area, internal standard method.

HPLC: Rp 18 column with H2O:CH3CN:H3PO4: HEXYLAMINE

Spectroscopic Analysis:Using IR and MS spectroscopic methods. Pure sample of

mescaline can be isolate from plant material or other illicit product by PTLC or

HPLC.

The major peaks in the IR spectrum of mescaline base, SO4 and HCl occur at the

following wave numbers (cm-1, 1591, 1513, 1245, 1130, 995, 835, 670).

The characteristic fragment ions in the EI MS of mescaline (m/z) 211, (molecular

ion), 182 (base peak), 167, 151.

N.B. UV is not specific for the analysis of mescaline because other

phenylethylamines yield similar results and therefore it is not recommended.

Catecholamine-Like Hallucinogens

Elemicin

2-Nutmeg

Myristicin: found in nutmeg

Elemicin: found in mace

Both Has similar structure as Mescaline

Action: Blocks release of serotonin in brain neurons.

Unreality Confusion Disorientation, Euphoria, Visual

hallucinations and Acute psychotic reactions

5-15g. (approximately 2 tablespoons) in tea will produce

confusion, disorientation, depersonalization, and feelings

of unreality

Side Effects: vomiting, nausea, agitation, dry mouth,

rapid heart beat, and tremors.

Negative effects can last up to 12 hours.

2-Nutmeg mace nutmeg

Psilocybin, Psilocin,

Bufotenine

LSD

Serotonin-like compounds

1-Psilocybe Mushroom

Hallucinogens


-Psilocybe mushrooms played a major role in the divinatory and magic rites of the

ancient inhabitants of the Aztec empire.

-Important religious cult us based on the sacramental consumption of their sacred

mushrooms also called divine flesh.

- Still today the hallucinogenic mushroom cult, namely the use of the species

psilocybe Mexicana, is deeply rooted in the nature tradition of Mexican Indians.

Genus Psilocybe (strophariaceae) is the most important being an almost cosmopolitan genus

of hallucinogenic mushrooms.

More than 140 species are known, 80 of them are known to contain psychotropic substances

Their main distribution is in the temperate region.

The species grow on the soil and on a variety of organic substances such as humus, dung, rootling

wood, peat and also on clumps of mosses.

The most important two species from the aspect of drug abuse are Psilocybe semilanceata and

Psilocybe urbensis.

.

Hallucinogens


Psilocybe semilanceata known as liberty caps is the most widespread psilocybin-

containing mushroom occurring in north and middle Europe, north America, Russia

and Australia. It grows very scattered to gregarious or rich soil, among grass, or field

near farmyards.

Psilocybe Mexicana is known to grow in southern Mexico and Guatemala, especially

in limestone regions, growing isolated or sparsely in moss or herbs along roadside,

humid meadows, and corn field and in the neighborhood of pine and oak forests.

Psilocybe urbensis: is known to grow in south and central America, south mexico,

west indies, Florida and southeastern Asia. It grows singly or in small groups usually

on dung or rich Pasteur soil.

Since a few years ago, a marked increase has been observed in the abuse, by ingestion

of the fresh or dried fruit-bodies of their potent psychoactive mushrooms which are

controlled in many countries

Hallucinogens

1-Psilocybe Mushroom Active constituents:

The main alkaloids present in these species are the phosphorylated indolamines

psilocybin (4-phosphoryloxy-N,N-dimethyl tryptamine) and beaocystin (4-phosphonyl-

oxy-N-methyl tryptamine), norpsilocybine, the latter being possibly the direct

biochemical precursor of psilocybin.

Psilocin (4-hydroxy-N,N-dimethyl-tryptamine), the desphosphorylation product and

hallucinogenic metabolite of psilocybin is usually present only in trace amount.

It is presently still not known whether baeocystin produces psilocybin-like

psychotropic effect.

Therefore psilocybin and psilocin are considered, from the aspect of drug of abuse, as

the chemical constituents of forensic significance.

Psilocybin, legally or illegally synthesized or isolated from mushroom, is from time to

time encountered in the illicit drug traffic.

Other psilocybin-containing mushrooms are belonging to the genera Panaeolus

(coprinaceae), Conocybe (Bolbitacea), Inocybe (Cortnariacea) and Pluteus (Plueaceae).

Hallucinogens


Pharmacological Effects: 1-After oral ingestion, psilocybin is converted through dephosphorylation to the more

lipophilic psilocin.

2-lipophilic psilocin is rapidly distributed through the body including CNS. It is duration

action is from 2-4 hours (much shorter than that of lysergic and mescaline).

3-Psilocybin produces characteristic physiological effects that precede the

psychological ones.

These may include the dilatation of pupils and increase body temperature and increase

blood pressure.

When taken in sufficient quantities, psilocybin and its more active metabolite

psilocin produce changes in the perceptions of one’s body (reportedly one’s spirit

seems to leave the body and take flight).

There is also distortion in time and space perception and hallucination similar to

those under lysergide.

Hallucinogens


Seizures of psilocybe mushrooms may consist of several fresh, dried or pulverized

fruit-bodies. Psilocybin exhibits mostly in its brown-yellow colored base form as

powders, tablets or capsules in a single containers or package.

Both products are potent hallucinogen after 30 minutes of oral admistration.

It can be analysis by three methods .a. morphological, b. coluer test,c.

chromatographic methods

a. Morphological (Macro- and microscopic characters):

Agarics traditionally have been identified after recording of only field characters such as

color and texture, which depends on not only the availability of fresh material but also

the presence of entire fruit-body.

In addition, many of the characters used are very subjective and variable.

Therefore the identification of psilocybe mushrooms basing on a macroscopic

examination needs some experience and should only reformed by trained mycologist.

Also the microscopic examination of fragments or powders of psilocybe mushrooms

specimens should only be performed by trained mycologists.

Illicit products and its analysis

Hallucinogens

1-Psilocybe Mushroom Illicit products and its analysis

b.Colors tests: Many other C2 unsubstituted indoles e.g. DMT and lysergide as well as those which are

harmless and uncontrolled by national legislation or international treaties may give similar

colors with the test reagents

Ehrlich reagent: 1 g P-dimethyl amino benzaldehyde is dissolve in 10 cc methanol, then add 10 ml

orthophosphoric acid. To the small amount of the suspected material (from powder, tablet or capsules).

Placed in spotting plate, +2 drops of reagent it will give violet to grey violet color.

Appearing after few minutes indicate the possible presence of psilocin (detection limit is 1 µg).

N.B. a preextraction step is necessary for color tests of psilocybe mushrooms, since the vegetable color

material can interfere.

Marqu’s Reagent: Solution1- 8-10 drops of 40% HCHO+ 10 ml of glacial acetic acid. Solution2-

H2SO4. To small amount of the suspected material (from powder, tablet or capsules) placed in spotting

plate and 1 drop of solution 1- and 2 drops of H2SO4. An orange color indicates the possible presence of

psilocybe while a green-brown color indicates the possible presence of psilocin. The detection limit is 10

µg.

N.B. color developed is interfered with vegetable material of mushrooms and because many other natural

products develop a yellow color with concentrated H2SO4, this color test is not recommended for the

mushroom material.

Hallucinogens


c. Chromatographic Analysis:

TLC: Sample preparation: For Psilocybe mushroom

20 mg powder

shaking

MeOH, 2 ml

For 5 minutes

Filter

evaporate

under nitrogen

concentrate alcohol

extract

•Psilocin is sparingly soluble in MeOH especially when exposed to light or air.

•For psilocybin tablets and capsules: Extract representative sample of the finely powdered

tablet or the content of capsules by shaking or sonicating with MeOH alcohol.

TlC: Is Spotted Alongside Standard Solution Of Psilocybin And Psilocin System:

a)N-butanol: Hoac: H2o (20:10:10), b) MeOH: NH4OH (100:1.5

Visualization: observe first the plate under UV light at 254 and 366 nm, then spray

with Erlich Reagent (P-DMAB+MeOH+HCl). Both Erlich and P-DMAB give gray-

violet to violet color with psilocybin and baeocytin and blue color with psilocin.

N.B. P-DMAB is more sensitive and yields better color stability.

Hallucinogens



GLC: direct injection of psilocybe mushroom extracts or psilocybin sample solution on

packed or capillary columns will convert psilocybin by thermal dephosphoriylation is necessary

if psilocybin is to be detected. Moreover, sugars are present in considerable amounts in

psilocybe mushroom extracts which may interfere with the derivatization. Elimination of sugar

is, therefore necessary before the preparation of derivative sample.

Without derivatization: For psilocybe mushroom extract 20 mg of the powder by shaking for 15

minutes with internal standard. For psilocybin powder of tablets or capsules extraction is like on TLC

Standard solution: add internal standard solution to an accurately weighed amount of

psilocybin is 1mg/ml. Internal standard: dissolve on appropriate n-alkane in MeOH to give a

concentration of 1mg/ml. Conditions: column fused silica capillary column carrier gas He,

detector FID. Quantitative: by peak area, internal standard method tR of psilocin 11.2

minutes.

With derivatization: Sample preparation: for psilocybe mushroom as above but to eliminate sugars add

1 ml of acetone to methanolic solution allow the mixture to stand for 30 minutes at room temperature or for

10 minutes in a freeze, centrifuge or filter the precipitated sugars evaporate under nitrogen then prepare

TMS derivative the rest of procedure as above for non derivatization.Results: psilocin-di-TMS, psilocybin-

tri-TMS and baeocystin-tri-TMS have retention time of 11.9, 15.3 and 15.7 minutes respectively.

Hallucinogens



HPLC: Normal Phase: Prepare the solution as outlined under GLC without derivatization

but without adding internal standard. Detectors UV, Flurometric, electronical or reference

electrode.Rp-8: Prepare Also As Above But Add Bufotenine As Internal Standard. Column:

Rp-8.Quantitation by peak area, internal standard method.

Spectroscopic analysis: IR and MS are used for identification of isolated or synthesized

psilocybin and psilocin. Depending on the purity of these compounds, an extraction step may

be required prior to spectroscopic analysis. In this case pure sample of psilocybin and psilocin

have to be isolated from psilocybe mushroom or other illicit products by PTLC using

nButanol: HoAC:H2O (BAW, 20:10:10).

Results: IR: psilocybin (KBr cm-1) 1620, 1585,1505, 1360, 1065.Psilocin (KBr cm-1) 1620,

1585, 1261, 1236, 1061, 1042 EIMS:-Psilocin (underivatized) m/z 204 (M+), 146, 136, 77, 58

(BASE PEAK), 42.Psilocin di-TMS m/z (M+, 384), 290, 73, 58 (Base peak).psilocybin-tri-

TMS m/z 500 (M+), 485,455, 442, 73, 58 (11)

N.B.:1-UV spectroscopy is not specific for the analysis of psilocybin and psilocin because other dimethyl tryptamine

derivatives yield similar results and therefore it is not recommended.

2-underivatized psilocybin and psilocin: cannot be differentiated by MS, because the thermal instability psilocybin is rapidly

converted to psilocin by thermal dephosphorylytion. Thus prior derivatization is necessary for their selective identification.

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Substance Abuse PHG 414 - PSAU · Hallucinogens 1-Peyote cactus Mescaline Tolerance : It builds...

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