{"id":510,"date":"2015-01-22T06:40:33","date_gmt":"2015-01-22T06:40:33","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=510"},"modified":"2018-10-06T04:58:17","modified_gmt":"2018-10-06T04:58:17","slug":"phytochemical-investigations-of-plant-trichodesma-indicum","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol1no2\/phytochemical-investigations-of-plant-trichodesma-indicum\/","title":{"rendered":"Phytochemical Investigations of Plant Trichodesma Indicum"},"content":{"rendered":"

Materials and Methods<\/strong>\u00a0<\/strong><\/p>\n

Collection and extraction of plant<\/strong><\/p>\n

The plant was collected in Mysore and taxonomical identification of the plant specimen was authenticated by Prof. Shashikala, HOD of Botany, Sarada Vilas Science College, Mysore. The plant was removed and dried under shade and powdered in a mechanical grinder.\u00a0 The powder was extracted with various solvents.\u00a0 All chemicals and reagents used are of analytical grade.<\/p>\n

Sample taken\u2014Trichodesma indicum<\/em> whole plant extract<\/p>\n

Solvent\u2014Petroleum ether,Benzene,Chloroform,Alcohol and Water.<\/p>\n

Method used\u2014Reflux method.<\/p>\n

Apparatus\u2014Heating mantle,beaker, round bottom flask,measuring cylinder, double walled condenser,stand etc.<\/p>\n

Proportion of\u00a0 the drug with the solvent\u20141:4<\/p>\n

Quantity of the drug taken 100g<\/p>\n

Time-16hrs<\/p>\n

Drying temperature—–<500<\/sup> C<\/p>\n

Extraction Procedure 1<\/sup><\/strong><\/p>\n

The preparation of various extracts was by successive solvent extraction. The material to be extracted was subjected to extraction with solvents in ascending order of polarity successively. The constituents which are soluble in both polar and non polar solvents can be extracted separately by adopting this approach. For each and every extraction the extract was evaporated under reduced pressure until the solvent had been removed to give an extract sample with a yield of w\/w.<\/p>\n

All the extracts were examined for their colour and consistency. Their percentage weights were calculated with reference to the dried sample. These data are compiled in table no.1\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/strong><\/p>\n

Table 1:<\/strong>\u00a0<\/strong><\/p>\n\n\n\n\n\n\n\n
Sl No.<\/strong><\/td>\nNa\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Name of the<\/strong><\/p>\n

extract<\/strong><\/td>\n

\u00a0\u00a0\u00a0\u00a0\u00a0 Nature<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 Colour<\/strong><\/td>\nWt. of\u00a0 Extract<\/strong><\/td>\n<\/tr>\n
1.<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Pet.Ether<\/td>\n\u00a0\u00a0\u00a0 Resinous<\/td>\nYellowish green<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 2g<\/td>\n<\/tr>\n
2.<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 Chloroform<\/td>\n\u00a0\u00a0\u00a0 Resinous<\/td>\nGreenish<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 2.5g<\/td>\n<\/tr>\n
3.<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Alcohol<\/td>\n\u00a0\u00a0\u00a0 Resinous<\/td>\nGreenish brown<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a04g<\/td>\n<\/tr>\n
4.<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Aqueous<\/td>\n\u00a0\u00a0\u00a0 Resinous<\/td>\nBrownish yellow<\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 3.5g<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Qualitative chemical tests for various chemical\u00a0\u00a0 Phyto constituents. <\/strong><\/p>\n

Qualitative tests were conducted for all the extracts to identify the various phyto constituents .The various tests for steroids, flavonoids and\u00a0 tannins were observed and the tests are recorded in table no.2 which shows the qualitative chemical\u00a0 tests conducted for various extracts.<\/p>\n

Table 2:<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\u00a0\u00a0 Sl.No.<\/strong><\/td>\nChemical test<\/strong><\/td>\nPet.Ether<\/strong><\/td>\n\u00a0\u00a0 CHCl3<\/sub><\/strong><\/td>\n\u00a0 Alcohol<\/strong><\/td>\n\u00a0Aqueous<\/strong><\/td>\n<\/tr>\n
1.<\/td>\nSteroids<\/td>\n+ve<\/td>\n+ve<\/td>\n-ve<\/td>\n-ve<\/td>\n<\/tr>\n
2.<\/td>\nTriterpenes<\/td>\n-ve<\/td>\n-ve<\/td>\n+ve<\/td>\n+ve<\/td>\n<\/tr>\n
3.<\/td>\nSaponins<\/td>\n-ve<\/td>\n-ve<\/td>\n+ve<\/td>\n+ve<\/td>\n<\/tr>\n
4.<\/td>\n.Alkaloids<\/td>\n-ve<\/td>\n-ve<\/td>\n-ve<\/td>\n-ve<\/td>\n<\/tr>\n
5.<\/td>\nFlavonoids<\/td>\n-ve<\/td>\n-ve<\/td>\n+ve<\/td>\n+ve<\/td>\n<\/tr>\n
6.<\/td>\nTannins<\/td>\n-ve<\/td>\n-ve<\/td>\n+ve<\/td>\n+ve<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Purification and separation of Flavanoidal compounds by column chromatography <\/strong>2<\/sup><\/strong><\/p>\n

Attempts were made to purify and separate the Flavanoidal compound by taking alcoholic extracts (prepared by successive solvent extraction) and mixed with silica gel and kept overnight to get adsorbed on the silica gel. Then the column was packed with silica gel using petroleum ether for building the column.\u00a0 Then the silica gel along with alcoholic extract was packed at the top of the column and the column was developed using different solvents. The solvent mixtures were collected in the decreasing order of their polarity as shown in the table no.3 :<\/p>\n

Different fractions were collected at the rate of 18 to 20 drops per minute and each fraction was subjected to TLC examination. The course of chromatogram is given below:<\/p>\n

Table 3:<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
 <\/p>\n

Sl.no<\/strong><\/td>\n

 <\/p>\n

Fractions<\/strong><\/td>\n

 <\/p>\n

Eluate<\/strong><\/td>\n

 <\/p>\n

Nature<\/strong><\/td>\n

 <\/p>\n

Color<\/strong><\/td>\n<\/tr>\n

 <\/p>\n

1.<\/td>\n

 <\/p>\n

1-4<\/td>\n

 <\/p>\n

PetEther(100%)<\/td>\n

 <\/p>\n

colorless<\/td>\n

 <\/p>\n

-ve<\/td>\n<\/tr>\n

 <\/p>\n

2.<\/td>\n

 <\/p>\n

5-10<\/td>\n

 <\/p>\n

Pet.Ether+Benzene<\/p>\n

(50:50)<\/td>\n

 <\/p>\n

V.light<\/p>\n

yellow<\/td>\n

 <\/p>\n

-ve<\/td>\n<\/tr>\n

 <\/p>\n

3.<\/td>\n

 <\/p>\n

11-15<\/td>\n

 <\/p>\n

Benzene+EtOH<\/p>\n

(75:25)<\/td>\n

light brownish yellow<\/td>\n <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

4.<\/td>\n

 <\/p>\n

16-20<\/td>\n

 <\/p>\n

Benzene+EtOH (50:50)<\/td>\n

 <\/p>\n

yellowish brown<\/td>\n

 <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

5.<\/td>\n

 <\/p>\n

21-25<\/td>\n

 <\/p>\n

Benzene+EtOH (25:75)<\/td>\n

 <\/p>\n

Light orange yellow<\/td>\n

 <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

6.<\/td>\n

 <\/p>\n

26-30<\/td>\n

 <\/p>\n

EtOH(100%)<\/td>\n

greenish yellow<\/td>\n <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

7.<\/td>\n

 <\/p>\n

31-37<\/td>\n

 <\/p>\n

Acetone+MeOH<\/p>\n

(50:50)<\/td>\n

brownish yellow<\/td>\n <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

8.<\/td>\n

 <\/p>\n

38-42<\/td>\n

 <\/p>\n

Acetone+MeOH<\/p>\n

(25:75)<\/td>\n

 <\/p>\n

light yellow<\/td>\n

 <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

9.<\/td>\n

 <\/p>\n

43-48<\/td>\n

 <\/p>\n

MeOH(100%)<\/td>\n

brownish yellow<\/td>\n <\/p>\n

+ve<\/td>\n<\/tr>\n

 <\/p>\n

10.<\/td>\n

 <\/p>\n

49-54<\/td>\n

 <\/p>\n

EtOH(100%)<\/td>\n

greenish yellow<\/td>\n <\/p>\n

+ve<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Examination of\u00a0the Eluates\u00a0<\/strong><\/p>\n

Fraction 1-10<\/strong>\u00a0<\/strong><\/p>\n

All the fractions showed \u2013ve\u00a0 test for Flavoniods .Hence further\u00a0 investigation was not carried out.<\/p>\n

Fraction 15-54<\/strong><\/p>\n

All the fractions gave +ve test for Flavoniods and subjected to TLC examination. These fractions showed a single spot having same Rf value. Hence all the fractions were pooled together and the solvents were removed under reduced pressure.<\/p>\n

The residue was dissolved in alcohol and left overnight. The light brownish yellow compound was obtained which was soluble in EtOH, Acetone, MeOH, dil H2<\/sub>SO4<\/sub> and DMSO.<\/p>\n

The isolated Flavonoidal compounds gave +ve test for the following color reactions:<\/p>\n

Fecl3<\/sub> test: Dark green color<\/p>\n

Magnesium ribbon Hcl reductiontest-Crimson red color (Shinoda test)<\/p>\n

Zinc-Hcl reduction test \u2013yellow ppt<\/p>\n

Ammonium Hydroxide solutes test-yellow flouresnce (UV)<\/p>\n

TLC study of isolated compounds<\/strong><\/p>\n

Materials used- Glass plate 20X5cm size<\/p>\n

Adsorbent- Silica gel-G<\/p>\n

Solvent system used- \u00a0\u00a0\u00a0a) Ethyl acetate: AA (9:1)<\/p>\n

b) Pet.Ether : ethanol (1:9)<\/p>\n

c) Toluene: Ethyl alcohol: acetone (7:2:2)<\/p>\n

Sample: \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Isolated flavonoids<\/p>\n

Detecting agent: \u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Antimony trichloride<\/p>\n

Rf value: \u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a00.9<\/p>\n

Further it needs spectral studies to confirm the findings. In a similar fashion steroids (obtained from Pet.Ether- CHCl3<\/sub> extract) were subjected to column chromatography and \u00a0fractions were tested for solvents collected at regular intervals of time and each fraction was subjected to TLC study which showed two different Rf values i.e, 0.9 & 0.75 with different solvent systems which are shown in the Table no.4<\/p>\n

Similarly triterpenoids were isolated by taking alcoholic extract with the help of column chromatography & fractions were collected, tested for triterpenoids and gave positive response and later it was subjected to TLC study. It has also shown two different Rf values (0.9& 0.72) with different nature of their residues. Further studies like UV, IR & Mass, NMR spectral studies are required to confirm the findings.<\/p>\n

Table 4:\u00a0TLC study of the various extracts of Trichodesma indicum.<\/strong><\/p>\n\n\n\n\n\n\n
Sl.No<\/td>\nChemical group<\/td>\nSolvent system<\/td>\nSpray\u00a0 reagent<\/td>\n <\/p>\n

Rf value<\/p>\n

 <\/td>\n<\/tr>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

1<\/td>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

Steroids<\/td>\n

a)\u00a0\u00a0\u00a0\u00a0\u00a0 CHCl3<\/sub>:MeOH (7:3)<\/p>\n

 <\/p>\n

b)\u00a0\u00a0\u00a0\u00a0\u00a0 Toluene:<\/p>\n

EtoH:diethylamine<\/p>\n

 <\/p>\n

 <\/p>\n

c)\u00a0\u00a0\u00a0\u00a0\u00a0 CHCl3<\/sub>:EtOH (7:3)<\/p>\n

 <\/td>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

Liebermann Burchard Reagent<\/td>\n

0.9<\/p>\n

 <\/p>\n

 <\/p>\n

0.9&0.75<\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

0.9&0.72<\/td>\n<\/tr>\n

 <\/p>\n

2<\/td>\n

\u00a0\u00a0 Triterpenoids<\/td>\n <\/p>\n

CHCl3<\/sub>: MEOH (7:3)<\/td>\n

 <\/p>\n

Antimony Trichloride Reagent<\/td>\n

 <\/p>\n

 <\/p>\n

0.87&0.72<\/td>\n<\/tr>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

3<\/td>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

Flavonoids<\/td>\n

 <\/p>\n

a) EA:HAc (9:1)<\/p>\n

 <\/p>\n

b)CHCl3<\/sub>: MeOH (1:1)<\/p>\n

 <\/p>\n

c)Toluene:<\/p>\n

EtOH:acetone (2:4:4)<\/p>\n

 <\/p>\n

d)EtOH: formic acid: HAc:water<\/p>\n

(100:11:11:26)<\/p>\n

 <\/p>\n

 <\/td>\n

<\/td>\n <\/p>\n

0.9,0.81&0.54<\/p>\n

 <\/p>\n

0.92&0.75<\/p>\n

 <\/p>\n

0.92&0.85<\/p>\n

 <\/p>\n

 <\/p>\n

0.93&0.86)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Discussion and\u00a0results<\/strong><\/p>\n

Trichodesma indicum <\/em>is a plant containing steroids, triterpinoidal saponins & Flavanoids.The qualitative tests & chromatographic studies substantiate the presence of Steroids, triterpenoidal saponins & flavanoids. The presence of these phytochemical constituents may have different biological activities.\u00a0 The Pharmacological screening of various extracts that are taken up is under progress in our laboratories.<\/p>\n

Acknowledgements<\/strong><\/p>\n

The authors thank Prof.Shashikala, H.O.D.,Dept.of Botany, SaradaVilas Science College, for taxanomical identification of the plant specimen. The authors wish to thank the management for encouraging and providing laboratory facilities to pursue this work.<\/p>\n

References<\/strong>\u00a0<\/strong><\/p>\n

    \n
  1. Kokate C.K.PurohitA.P.,GokhaleS.B.,2004,Pharmacognosy In.,PhytochemicalInvestigations,twentysixth ed.,Nirali Prakashan,Pune, Mumbai,pp105-109.<\/li>\n
  2. G.Anand S.,1996,Instrumental methods of Chemical Analysis,Himalaya Publishing House,Mumbai,P 585.<\/li>\n
  3. E.,2005,Thin Layer Chromatography,A laboratory Hand Book,second ed,Springer-Verlag,Berlin,Heidelberg,Newyork,Pp52-71.<\/li>\n
  4. E.,2005,Thin Layer Chromatography,A laboratory Hand Book,second ed,Springer-Verlag,Berlin,Heidelberg,Newyork,Pp125-133.<\/li>\n
  5. E.,2005,Thin Layer Chromatography,A laboratory Hand Book,second ed,Springer-Verlag,Berlin,Heidelberg,Newyork,Pp316-324.<\/li>\n
  6. Wagner H.,Bladt,S.,2002,Plant Drug Analysis In,Thin Layer Chromatography,second edn, Springer-Verlag, Heidelberg,Germany,P335<\/li>\n
  7. Kirtikar,K.R.,Basu,B.D.,2000,Indian medicinal plants,SriSatguru Publications,Delhi,Pp2335-37.<\/li>\n
  8. Vinod .D.Pharmacognosy & Phytochemistry,2006,Part-I,In,Characteristics for Citrous Bioflavanoids,First ed,Carrier Publications,Nasik,Maharashtra,P338<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Materials and Methods\u00a0 Collection and extraction of plant The plant […]<\/p>\n","protected":false},"author":2,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[5],"tags":[],"class_list":["post-510","post","type-post","status-publish","format-standard","hentry","category-vol1no2"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/510","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\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=510"}],"version-history":[{"count":8,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/510\/revisions"}],"predecessor-version":[{"id":13253,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/510\/revisions\/13253"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=510"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=510"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=510"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}