{"id":516,"date":"2015-01-22T06:25:07","date_gmt":"2015-01-22T06:25:07","guid":{"rendered":"http:\/\/biomedpharmajournal.org\/?p=516"},"modified":"2020-04-25T06:32:55","modified_gmt":"2020-04-25T06:32:55","slug":"a-flavonoid-out-of-tephrosia-purpurea-extract-and-its-antimicrobial-effect","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol1no2\/a-flavonoid-out-of-tephrosia-purpurea-extract-and-its-antimicrobial-effect\/","title":{"rendered":"A Flavonoid out of Tephrosia Purpurea Extract and its Antimicrobial Effect"},"content":{"rendered":"

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

The production of free radicals (reactive oxygen containing molecule) in all in energy producing mechanism result in oxidative stress linked with cancer, ageing, atherosclerosis, inflammation and neurodegenerative diseases. Flavonoids are actively involved in the detoxification of highly reactive species.<\/p>\n

Detoxification of hydrogen peroxide in vacuole has been proposed by Yamaski H in 1997.<\/p>\n

2 FlaOH\u00a0\u00a0 +\u00a0\u00a0\u00a0\u00a0 H2O2\u00a0\u00a0\u00a0\u00a0 =\u00a0\u00a0\u00a0\u00a0\u00a0 2 Flavo*\u00a0\u00a0\u00a0 +\u00a0\u00a0\u00a0 2 H2O<\/p>\n

Flavonoid Peroxidase\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0 Flavonoid phenoxyl radical<\/p>\n

The resulting flavonoid Phenoxyl radical can react with ascorbic acid (ASA) to generate the monodehydroascorbic acid radical (MDA*).<\/p>\n

2 Flavo*\u00a0\u00a0\u00a0 +\u00a0\u00a0\u00a0\u00a0 2 ASA\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 =\u00a0\u00a0 2 FlavOH\u00a0\u00a0\u00a0 + 2 MDA*<\/p>\n

MDA* can in turn nonenzymatically generate ASA and DHA (Dehydorascorbate reductase).<\/p>\n

Tephrosia purpurea<\/em> of family Leguminosae are very rich in flavonoid. It is a small annual shrub used in the treatment of asthma. Roots are useful to cure cuts and wounds. Whole plant is used in spleen enlargement and stem used as tooth brush cure many dental disorder. It cures colic pain when used with sendha salt.<\/p>\n

Flavonoids are synthesized by the phenylpropanoid metabolic path way in which the amino acid phenylalanine is used to produce 4-coumaroyl-CoA.<\/p>\n

Material and Method<\/strong><\/p>\n\n\n\n
\"Figure<\/td>\nFigure 1:<\/strong><\/p>\n

 <\/p>\n

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

 <\/p>\n

Plant Material<\/strong><\/p>\n

The plant material (near about 2 Kg) collected from local surroundings after weighing, washed thoroughly and dried in air at room temperature for more than one month. The dried plant material was grinded to powder to about 40-60 mesh size. Then it was weighted and stored carefully in bottles for extraction and chemical testing.<\/p>\n

Extraction and Isolation<\/strong><\/p>\n

Extraction was carried out in Soxhlet apparatus in the laboratory with 90% ethanol as solvent. The extract was concentrated in vaccum evaporator to a good yield of 6.3%.<\/p>\n

The biologically active compound was separated from the crude extract by column chromatography using EtoAc: ccl4 solvent system as mobile phase and silica gel as stationary phase.<\/p>\n

The compound was than isolated followed by TLC using same solvent combination for that fraction in column.<\/p>\n

Table 1:\u00a0showing the weight of plant material after drying and percentage loss in weight of plant material of Tephrosia purpuria.<\/em><\/strong><\/p>\n\n\n\n\n\n\n\n
S. No.<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Description<\/strong><\/td>\nWeight in <\/strong><\/p>\n

\u00a0\u00a0\u00a0\u00a0 gms.<\/strong><\/td>\n<\/tr>\n

1.<\/td>\nWt. Of plant material in wet fresh condition<\/td>\n240 gms.<\/td>\n<\/tr>\n
2.<\/td>\nWt. Of plant material after drying condition<\/td>\n160 gms.<\/td>\n<\/tr>\n
3.<\/td>\nLoss in weight on drying<\/td>\n240\u2013160 = 80 gms.<\/td>\n<\/tr>\n
4.<\/td>\nPercentage loss after drying<\/td>\n40%<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

Table: 2 Percentage yield of Tephrosia purpuria <\/em>using different solvents in Soxhlet apparatus.<\/strong><\/p>\n\n\n\n\n\n
S.No.<\/strong><\/td>\nSolvent <\/strong><\/p>\n

\u00a0\u00a0 used<\/strong><\/td>\n

Wt. of\u00a0 Of Powdered<\/strong><\/p>\n

Material<\/strong><\/td>\n

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

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

\u00a0\u00a0 Solvent<\/strong><\/td>\n

Wt. of <\/strong><\/p>\n

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

Character <\/strong><\/p>\n

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

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

Percentage<\/strong><\/p>\n

of yield<\/strong><\/td>\n<\/tr>\n

1.<\/td>\n90% Ethanol<\/td>\n160 gms.<\/td>\n750 ml.<\/td>\n7.20 ml.<\/td>\nGreen in<\/p>\n

semisolid<\/p>\n

state<\/td>\n

6.3%<\/td>\n<\/tr>\n
2.<\/td>\nWater<\/td>\n160 gms.<\/td>\n500 ml.<\/td>\n8.00 ml.<\/td>\nGreenish<\/p>\n

yellow<\/p>\n

semisolid<\/p>\n

state<\/td>\n

7.5%<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

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

Table: 3 \u00a0 TLC of 90% alcoholic extract of the plant Tephrosia purpuria.<\/em><\/strong><\/p>\n\n\n\n\n\n
S.No.<\/strong><\/td>\nSolvent System<\/strong><\/td>\nRatio<\/strong><\/td>\nVisual <\/strong><\/p>\n

light<\/strong><\/td>\n

Iodine<\/strong><\/p>\n

Chamber<\/strong><\/td>\n

Rf Value<\/strong><\/td>\nFraction <\/strong><\/p>\n

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

1.<\/td>\nEtoAc: CCl4<\/sub><\/p>\n

 <\/td>\n

1:1<\/td>\nBlackish<\/p>\n

green<\/td>\n

Dark<\/p>\n

Green<\/td>\n

0.92<\/td>\nFR – 1<\/td>\n<\/tr>\n
2.<\/td>\nCHCl3<\/sub>: MeOH<\/p>\n

 <\/td>\n

2:1<\/td>\nOlive<\/p>\n

green<\/td>\n

Dark<\/p>\n

Green<\/td>\n

0.86<\/td>\nFR – 2<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

 <\/p>\n

Observation and Results<\/strong><\/p>\n

The tentative structure of compound was elucidated as Mf. C21<\/sub>H20<\/sub>O11<\/sub>.<\/p>\n\n\n\n
\"Formula<\/td>\nFormula 1:<\/strong><\/p>\n

 <\/p>\n

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

 <\/p>\n

Molecular Formula -3, 7, 2\u2019 \u0301 trihydroxy 6\u2019 \u0301 \u2013 O \u2013 <\/strong>\u03b2<\/strong> \u2013 D glucopyranosyl flavone<\/strong><\/p>\n

Molecular Formula \u2013 C21<\/sub>H21<\/sub>O11<\/sub><\/strong><\/p>\n

Molecular Weight \u2013 449<\/strong><\/p>\n

M.P. \u2013 210 \u2013 212 0<\/sup>C<\/strong><\/p>\n

It was light greenish amorphous powder separated as fraction-1 from column. It was blackish green colored compound with Rf value 0.92 on TLC was followed by acid hydrolysis, methylation and glycosidal test.<\/p>\n

IR<\/strong><\/p>\n

3524 (OH), 1650 (C = O \u2013 \u03b3 \u2013 pyrone), 1590, 1565, 1515 (C = C aromatic)<\/p>\n

PMR<\/strong><\/p>\n

(<\/strong>Py) -8.35 (3H, d, J = 9.6, H \u2013 5, 2\u2019, 6\u2019), 7.15 (1 H, br. s, 3 \u2013 OH), 7.44 (4H, m, aromatic protons) 10.30 (1H, br. s, 7 \u2013 OH), 9.80 (1H, br. s, 4\u2019 – OH), 13.62 (1H, s, 3 – OH).<\/p>\n

On analysing under different analytical procedure showed maximum peak in ultraviolet spectrum at 270 mm.<\/p>\n

\u00a0Table 4:\u00a0Chemical Properties of Tephrosia purpurea<\/strong><\/p>\n\n\n\n\n
S.No.<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Properties<\/strong><\/td>\n\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 Percentage<\/strong><\/td>\n<\/tr>\n
1.<\/p>\n

2.<\/p>\n

3.<\/p>\n

4.<\/p>\n

5.<\/td>\n

Ash content<\/p>\n

Water soluble ash content<\/p>\n

Acid soluble ash content<\/p>\n

Fiber value<\/p>\n

Saponification value<\/td>\n

\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 30<\/p>\n

70<\/p>\n

4.2<\/p>\n

Not more than 30<\/p>\n

68<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n


\nDiscussion<\/strong><\/p>\n

The known compound 3,7,2\u2019 \u2013 trihydroxy 6\u2019- O – \u03b2 \u2013 D glucopyranosyl flavone was identified by CoTLC with authentic sample and structure was established by comparison of measured UV, NMR and mass spectral data with spectroscopic data available from literature. 3,7,2\u2019 \u2013 Trihydroxy 2\u2019- O – \u03b2 \u2013 D glucopyranosyl flavone showed 1<\/sup>H and 13CNMR data in full agreement with those given by\u00a0 Malikov and Yuldashev (2002).<\/p>\n

The chemical structure of flavanonols includes 3 \u2013 Hydroxyflavanones or Dihydroflavonols. They contain two asymmetric carbon atoms. Flavanols differed from flavanones by the presence of strong peaks in their mass spectra. It is demonstrated that flavanonols exhibit a base peak for [M \u201357] +<\/sup> ion whereas a peak for the [M \u2013 43] +<\/sup> ion was typical of flavones. It should be noted that an alcoholic solution of flavanonols at room temperature can undergo cis-trans epimerization to form a new isomeric compound.<\/p>\n

Several such compounds have been isolated from plants of different families.Genistein and Daidzein soy flavonoid appear to reduce the risk of prostrate and breast cancer, Hesperidins raises the blood levels of the good high-density lipoprotein and lowers the bad low-density lipoprotein and triglycerides. It also possesses significant anti-inflammatory and analgesic effect.<\/p>\n

Quercetin can inhibit the growth of head and neck cancers and can stop reverse transcriptase, the method HIV uses to replicate itself. It has demonstrated significant anti-inflammatory because of direct inhibition of several initial process of inflammation. As it inhibits both the production and release of histamine and other allergic inflammatory mediators. Tangeretin induces apoptosis programmed cell death in leukemia cells but does not harm normal cells Prenylnaringenin are very potential antioxidant. Epicatechin improves blood flows and thus seems good for cardiac cells.<\/p>\n

The flavon compound is isolated from Tephrosia Purpurea<\/em> was found to be anti-microbial when tested in experimental rats. A detail study of this compound is needed to be established.<\/p>\n

\u00a0<\/strong>Biological activity<\/strong><\/p>\n

The flavone isolated from alcoholic extract of stems and leaves of Tephrosia<\/em> purpurea <\/em>showed antimicrobial activity against Staphylococcus aureus. The bacterial strain used for the present study was procured from M.T.C.C. Chandigarh.\u00a0<\/strong><\/p>\n

Table 5:\u00a0Minimum inhibitory concentration (MIC) value of different fraction of flavonoids compound of Tephrosia purpurea against Staphylococcus aureus MTCC 96.<\/strong><\/p>\n\n\n\n\n
S.No.<\/strong><\/td>\nFraction<\/strong><\/td>\nMIC<\/strong><\/td>\n<\/tr>\n
1.<\/p>\n

2.<\/td>\n

\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 FR- 1<\/p>\n

FR-2<\/td>\n

\n

0.72<\/p>\n

0.34<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n

For antimicrobial activity disc diffusion was followed. The agar medium was used. The plates were incubated at 37 0<\/sup>C over night and examined for zones of growth inhibition.<\/p>\n

The minimum inhibitory concentration of the extract was determined for Staphylococcus aureus using the two fold serial micro dilution method with saline at a final concentration ranging from 10,000 mg \/ ml. to 0.0024 mg \/ ml. The MIC values were taken as the lowest concentration of the extracts in the wells of the microtiter plate. These plates were interpreted as visible growth of microorganisms. Thus growth of bacteria is noticed inhibited with moderate zone of inhibition.<\/p>\n

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

    \n
  1. Chaudhary Rashmi (2005), have reported ant asthmatic activity in alcoholic extract of Tephrosia purpurea. PhD thesis B.U., Bhopal – 221.<\/li>\n
  2. Malikov and Yuldashev (Chemistry of Natural compound vol. \u2013 38 No. – 5)<\/li>\n
  3. Guido flamini, Maria pardini, Ivano Morelli, Kuddisi Ertugrul (2002) Phytochemistry 61, pp. \u2013 433 \u2013 437.<\/li>\n
  4. Koli M.C., R. Chaudhary, Sourabh Kumar, Deepti Shukla and R.C. Saxena (2002). An isoflavonol glycoside from the stem of Euphorbia hirta<\/em> (Asian Journal of Chemistry vol. \u2013 14 No. \u2013 3, 1668-1673)<\/li>\n
  5. Kamgang R, Vidal, Poukam Kamgne E, I. Fonkoua Penlap N Beng V, Bivole Sida M: Activities of aqueous extract of Mallotus oppositi folium on Shigella dysenteriae. An induced diarrhea in rat clin Exp Pharmocol Physiol 2006; 33; 89-94.<\/li>\n
  6. Sashidharan, L. yoga, Latha Z. Zuraini, S. Suryani, S. Sangetha, L. Shirbi: Indian Journal of Pharmacology, vol. \u2013 39, October 2007, issue 5.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

    Introduction The production of free radicals (reactive oxygen containing molecule) […]<\/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-516","post","type-post","status-publish","format-standard","hentry","category-vol1no2"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/516","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=516"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/516\/revisions"}],"predecessor-version":[{"id":32927,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/516\/revisions\/32927"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=516"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=516"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=516"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}