{"id":41566,"date":"2021-12-30T10:28:23","date_gmt":"2021-12-30T10:28:23","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=41566"},"modified":"2022-01-04T08:11:22","modified_gmt":"2022-01-04T08:11:22","slug":"pharmacognostic-and-phytochemical-screening-of-datura-stramonium-by-tlc-and-gc-ms-a-forensic-approach","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol14no4\/pharmacognostic-and-phytochemical-screening-of-datura-stramonium-by-tlc-and-gc-ms-a-forensic-approach\/","title":{"rendered":"Pharmacognostic and Phytochemical Screening of Datura stramonium by TLC and GC-MS: A Forensic Approach"},"content":{"rendered":"

Introduction<\/strong><\/p>\n

Forensic Botany deals with the study of the plant science and is derived as botanical evidences in court of law for criminal administration purposes. From the historical point of view, plants and its extracts has been used as important tool as modus operandi in crimes such as, human poisoning, cattle poisoning and cattle revenge etc. Such type of botanical evidences can be found in suicidal, homicidal and accidental poisoning1<\/sup>.\u00a0 The plant derived botanical evidences may be Pollens grains, leaves, stem root, sap, resins\u00a0 and others encountered during crime scene investigations.<\/p>\n

Forensic Botany helps in linking the location, type of crime, revealing modus Operandi, culprit as well as victim by various circumstantial facts.\u00a0 There are various toxic plants grow\u00a0 very easily as\u00a0 aggressive\u00a0 weed in the temperate regions such as Datura stramonium, Abrus preactorious, Argemmone mexicana, Jatropha curcas, Ricinus communis, Nerium oleander etc.\u00a0\u00a0 In the present study, Datura stramonium has been considered as major plant encountered in poisoning cases in criminal Investigations2<\/sup>.<\/p>\n

Datura stramonium also called as Jimson weed, angels trumpet, thorn apple, and devil\u2019s trumpet and many other names is an annual poisonous plant. Its seeds, leaves, roots almost whole plant is poisonous and contains various poisonous compounds like atropine, hyoscyamine, and scopolamine3<\/sup>.<\/p>\n

Taxonomical Features of Datura<\/strong><\/p>\n

Datura stramonium belongs to family solanaceae, a nightshade family. Its origin is basically in North America.\u00a0 Its name was given by Carl Linnaeus also known as father of taxonomy. Leaves of it are coarse wavy at margin and are generally broad in length. Roots are long branched and thick4<\/sup>. The fruit of Datura stramonium is oval in shape and thorny in nature which contains many dark brown seeds. Flower of the plant is funnel shaped and generally white in colour.<\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig1\"<\/a><\/td>\nFigure 1: Fresh Leaves and Stem of Datura Stramonium<\/strong><\/p>\n

Click here to view figure\u00a0<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Toxicity of Datura Plant <\/strong><\/p>\n

The Datura plant is very toxic in nature therefore the chances of poisoning is more abundant during criminal investigations5<\/sup>. The Datura plant has toxic components in its all parts like leaves, seeds, stem and flowers6<\/sup>. The main toxic components of the Datura are scopolamine\/ Hyoscine, Atropine7<\/sup>. \u00a050\/100 seeds contain 3-6mg of atropine and each seed contains 0.1mg of atropine 8<\/sup>.<\/p>\n

Pharmacognostic effects of Hyoscine\/ Scopolamine<\/strong><\/p>\n

When hyoscine\/scopalamine as main constituent is administered in body has some adverse effects such as Nausea, Headache, unconsciousness, ulceration, high blood pressure, unable to breath, weakness, fatigue, are some common early symptoms9<\/sup>. Long effects such as Tachycardia, Dementia, Arrhythmia, Urinary retention, Blurred vision and dry mouth drowsiness10<\/sup>.<\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig2\"<\/a><\/td>\nFigure 2: Dried Fruit of Thorn Plant Containing Seeds Structure of Hyoscyamine<\/strong><\/p>\n

Click here to view figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Materials And Methods<\/strong><\/p>\n

Collection of Plant<\/strong><\/p>\n

The plant Datura stramonium fresh leaves were collected from Jhansi city of Bundelkhand region 25.4484\u00b0 N Longitude and 78.5685\u00b0 E Latitude at normal 300<\/sup>C and Wind N at 3 km\/h, and 80% Humidity with help of weather vane and hygrometer in the month of May. The plant was identified and described taxonomy in Department of Botany, Bundelkhand University, Jhansi. In the present study, the fresh leaves were washed to remove dirt and stored at 40<\/sup>C in aluminium foil. The following examinations were done.<\/p>\n

Microscopic and Macroscopic examination<\/strong><\/p>\n

Fresh leaves and stem of the plant was considered for the microscopic examination for species identification of the plant found in Bundelkhand region. Physical parameters of leaf, stem of plant were noted for species Identification.<\/p>\n

Phytochemical study of plant<\/strong><\/p>\n

Fresh leaves of Datura plant were crushed through pestle and motor method and filtered by muslin cloth and then by Whatman filter paper. Prepared extract was used for used for phytochemical study to identify the presence of phytochemicals11<\/sup>. Phytochemical study of plant were analysed for presence of alkaloids, sponins, anthraquinones, flavonoids, glycosides. Presence of glycosides and other highly poly complex structures may show the toxicity of the plant through phytochemical study12<\/sup>.<\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig3\"<\/a><\/td>\nFigure 3: Crushing by Pestle and Motar and Extract of leaves in glass bottle<\/strong><\/p>\n

Click here to view figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Preparation of extracts-The extract of plant collected after filtration from muslin cloth, extract was dissolved uniformly in two different solvents such as N-Butanol and N-Hexane for Phytochemical screening. Different chemical test were performed<\/p>\n

Chromatographic methods<\/strong><\/p>\n

TLC technique was applied for preliminary identification of toxic compounds such as Scopolamine\/ Hyoscyamine13<\/sup>. Atropine as major constituents for toxicity of plant. In this study hyoscine was detected through thin layer chromatography TLC.\u00a0 Hyoscyamine-N- butyl bromide standard Rf value was compared with the extracts of the plant leaves14<\/sup>. In different solvent\u00a0systems, chloroform: ethanol: water (81:11:8), chloroform: ethanol (3:1)15<\/sup>. The TLC plate was runned after spotting. Dragendroff reagent was used for visualization of alkaloids on the plate and Rf values were caluculated16<\/sup>.<\/p>\n

GCMS Data Analysis<\/strong><\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig4\"<\/a><\/td>\nFigure 4: The model (PerkinElmer Clarus 680) GC-MS was used for chromatographic analysis.<\/strong><\/p>\n

Click here to view figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table 1: The Acquisition Parameters Used in Operating the GC-MS.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Serial No <\/strong><\/td>\nOperation<\/strong><\/td>\nRunning Information<\/strong><\/td>\n<\/tr>\n
1<\/td>\nOven<\/strong><\/td>\nInitial temp 40\u00b0C for 5 min<\/td>\n<\/tr>\n
2<\/td>\nRamp<\/strong><\/td>\n12\u00b0C\/min to 260\u00b0C<\/td>\n<\/tr>\n
3<\/td>\nHold<\/strong><\/td>\n10 min<\/td>\n<\/tr>\n
4<\/td>\nInjection B Auto<\/strong><\/td>\n250\u00b0C<\/td>\n<\/tr>\n
5<\/td>\nVolume<\/strong><\/td>\n0 \u03bcL<\/td>\n<\/tr>\n
6<\/td>\nSplit<\/strong><\/td>\n50:1<\/td>\n<\/tr>\n
7<\/td>\nCarrier Gas<\/strong><\/td>\nHe<\/td>\n<\/tr>\n
8<\/td>\nSolvent Delay<\/strong><\/td>\n2.00 min<\/td>\n<\/tr>\n
9<\/td>\nTransfer Temp<\/strong><\/td>\n180\u00b0C<\/td>\n<\/tr>\n
10<\/td>\nSource Temp<\/strong><\/td>\n200\u00b0C,<\/td>\n<\/tr>\n
11<\/td>\nColumn<\/strong><\/td>\n30.0m x 250\u03bcm<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Results<\/strong><\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig5\"<\/a><\/td>\nFigure 5: A. Abaxial and adaxial View of Leaf and B. Microscopic View of Leaf.<\/strong><\/p>\n

Click here to view figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table 2:\u00a0The Microscopic Properties of Plant<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
Properties<\/strong><\/td>\nObservation<\/strong><\/td>\n<\/tr>\n
Color<\/td>\nAsh Green<\/td>\n<\/tr>\n
Shape<\/td>\nBroadly palmate<\/td>\n<\/tr>\n
Size<\/td>\n10-15 cm. x 7.5-12.5 cm<\/td>\n<\/tr>\n
Apex<\/td>\nPointed<\/td>\n<\/tr>\n
Venation<\/td>\nReticulate<\/td>\n<\/tr>\n
Margin<\/td>\nCordate<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Table 3:\u00a0Phytochemical Screening of Plant in Different Extracts<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
Serial No.<\/strong><\/td>\nChemical Test<\/strong><\/td>\nN- Butanol Extract<\/strong><\/td>\nN- Hexane Extract<\/strong><\/td>\n<\/tr>\n
1<\/td>\nAlklaoids<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
2<\/td>\nAmino Acid<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
3<\/td>\nAnthraquinones<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
4<\/td>\nGlycosides<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n
5<\/td>\nSponins<\/td>\n–<\/td>\n–<\/td>\n<\/tr>\n
6<\/td>\nFlavonoids<\/td>\n+<\/td>\n+<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note- Sign + {Present}, Sign- {Absent}<\/p>\n

The TLC chromatogram obtained after the calculation of Rf value and was matched with standard value {0.81}.The extract shows the presence of hysocyamine. Hyoscyamine is most lethal component found in Datura stramonium.<\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_gra1\"<\/a><\/td>\nGraph 1: shows the data of peaks obtained through GC of Datura stramonium<\/strong><\/p>\n

Click here to view Graph<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Interpretation of GC-MS Chromatogram<\/strong><\/p>\n

Table 4<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Serial No.<\/strong><\/td>\nRT Peak<\/strong><\/td>\nPeak %<\/strong><\/td>\nCompounds Identified<\/strong><\/td>\n<\/tr>\n
1.<\/td>\n4.995<\/td>\n1.505<\/td>\nHematoporphyrin, Tungsten, Aconitane-1<\/td>\n<\/tr>\n
2.<\/td>\n6.821<\/td>\n1.307<\/td>\nHematoporphyrin<\/td>\n<\/tr>\n
3.<\/td>\n9.912<\/td>\n2.365<\/td>\nO-methyloxime Delsoline, Methanesulfonic acid,<\/td>\n<\/tr>\n
4.<\/td>\n12.915<\/td>\n2.770<\/td>\n3-Pyridinecarboxylic acid, Hematoporphyrin Lycophyll, Rhodopin<\/td>\n<\/tr>\n
5.<\/td>\n16.118<\/td>\n3.572<\/td>\nDecanoic acid, Docosanoic acid,<\/td>\n<\/tr>\n
6.<\/td>\n18.867<\/td>\n1.603<\/td>\nTrilinolein, L-Proline, 5,5′-bis(trimethylstannyl)<\/td>\n<\/tr>\n
7.<\/td>\n21.703<\/td>\n1.004<\/td>\n\u00a0Octadecane, 3-ethyl-5-(2-ethylbutyl),\u00a0 Lanosta-7,9(11)-dien-18-oic acid, Tetracosane, 12-decyl-12-nonyl-<\/td>\n<\/tr>\n
8.<\/td>\n21.770<\/td>\n1.168<\/td>\nCholestano[7,8-a]cyclobutane, 3-methoxy-6-oxo-2′-methylene-, Carotene, 3′,4′-didehydro-1′,2′-dihydro-1′,2′-dihydroxy-, (2’R), Cinobufotalin<\/td>\n<\/tr>\n
9.<\/td>\n21.888<\/td>\n1.061<\/td>\nD-Glucopyranoside, Tungsten, 1,2-<\/p>\n

bis(dimethylphosphino)ethane<\/td>\n<\/tr>\n

10.<\/td>\n22.051<\/td>\n1.086<\/td>\ncis-Vaccenic acid, Octadecenoic acid, cis-13-Eicosenoic acid, cis-10-Nonadecenoic acid<\/td>\n<\/tr>\n
11.<\/td>\n23.016<\/td>\n2.509<\/td>\nAtropine, O-Bromoatropine, Hyoscyamine, Atropine, acetate<\/td>\n<\/tr>\n
12.<\/td>\n23.502<\/td>\n1.233<\/td>\nOleic acid, eicosyl ester, 17-Pentatriacontene, 9-Octadecene, 1-[2-(octadecyloxy)ethoxy]-<\/td>\n<\/tr>\n
13.<\/td>\n24.008<\/td>\n2.232<\/td>\nScopolamine, Benzeneacetic acid, Cyclopropanebutyric acid,<\/td>\n<\/tr>\n
14.<\/td>\n24.780<\/td>\n1.157<\/td>\n2-Nonadecanone 2,4-dinitrophenylhydrazine, Pentatriacontene, Tetrapentacontane, 1,54-dibromo-, 3-Desoxo-3,16-dihydroxy-12-desoxyphorbol 3,13,16,20-tetraacetate<\/td>\n<\/tr>\n
15.<\/td>\n25.891<\/td>\n2.355<\/td>\nRhodoxanthin, 4′-Apo-\u00e1,.psi.-carotenoic acid<\/td>\n<\/tr>\n
16.<\/td>\n28.918<\/td>\n1.108<\/td>\n2,4,6-Decatrienoic acid, Aconitane, trimethylsilyl<\/p>\n

ester,<\/td>\n<\/tr>\n

17.<\/td>\n29.053<\/td>\n1.562<\/td>\nCholestane, 3,5-dichloro-6-nitro-, 6-methyl-hept-5-<\/p>\n

enoic acid, methyl ester<\/td>\n<\/tr>\n

18.<\/td>\n29.348<\/td>\n1.880<\/td>\nD-Glucopyranosiduronic acid, Cholan-24-oic acidMethyl cholate<\/td>\n<\/tr>\n
19.<\/td>\n29.457<\/td>\n1.397<\/td>\n1,3-Dichloro-1,3-bis(norbornadien-2-yl)-1,3-bis(3-<\/p>\n

trimethylsilylpropyl)disiloxane, 9-Desoxo-9-x-acetoxy-3,8,12-tri-O-acetylingol<\/td>\n<\/tr>\n

20.<\/td>\n29.635<\/td>\n1.064<\/td>\ndicarbonyl-(\u00fc-4-2-methylenecycloheptanone)[1,2-<\/p>\n

bis(dimethylphosphino)ethane], 1′,1′-Dicarboethoxy-1\u00e1,<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_fig6\"<\/a><\/td>\nFigure 6: Structure of hyoscyamine (C17H23NO3)\/ Daturine identified in the extract of Datura leaves by GC-MS at RT 23.016 with 2.509 peak %.<\/strong><\/p>\n

Click here to view figure<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"Vol14No4_Pha_Bal_gra2\"<\/a><\/td>\nGraph 2: Shows MS spectrum of (RT Peak 23.016) m\/z for identification of Hyoscyamine compound with data reference from NIST Library and main library of GCMS.<\/strong><\/p>\n

Click here to view graph<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Conclusion<\/strong><\/p>\n

The Phytochemical screening of plant in different extracts showed the presence of alkaloids, saponins, flavonoids and amino acids. The alkaloid spots were identified by TLC. The spectral data of GC-MS confirmed the presence of different compounds such Atropine, scopolamine, hyoscyamine and their chemical characterization. This study substantiate with the previous literature that the leaves of Datura stramonium are toxic in nature. In cases of poisoning by Datura stramonium in Bundelkhand region, the plant can be easily identified through botanical derived evidences such as leaf traces or plant extracts during the investigations of crime scene and is very fatal (1471 ug\/kg) in men and eventually cause death, and contains high concentration of \u00a0major tropane alkaloids as hyoscyamine and scopolamine.<\/p>\n

Conflict of Interest<\/strong><\/p>\n

There is no conflict of interest including honorarium, grants, membership, employment, ownership of stock or any other interest or non\u2010financial interest such as personal or professional relation, affiliation and knowledge of the research topic between the authors.<\/p>\n

Funding Sources<\/strong><\/p>\n

There are no funding sources for this article.<\/p>\n

References<\/strong><\/p>\n

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    Introduction Forensic Botany deals with the study of the plant […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[94],"tags":[],"class_list":["post-41566","post","type-post","status-publish","format-standard","hentry","category-vol14no4"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/41566","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=41566"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/41566\/revisions"}],"predecessor-version":[{"id":42618,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/41566\/revisions\/42618"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=41566"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=41566"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=41566"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}