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Medicinal plant has pro vid ed the medical field with varies amount of new molecule.

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The use of plant as medicin e has being in practice since before 3000 B .C , since then

medicinal plants has been docume nted by each country throughout history (Raskin &

Ripoll 2004 ). The extended histo ry of medicinal plant is a good indication that plant

do possess the potential to provide large amount of new chemical entity to fight th e

outbreak of disease in the wor ld.

Further evidence of its significa nce according to

(Ashour, et al., 2013) half of th e best -selling pharmaceuticals in 1991 were either

from natural products or derivative analogues . Medicinal plant is preceded important

in developing countr ies , for its primary health care , due to its acceptability culturally ,

compati bility with the human body but also its contains less side effect compared to

the synthe tics that are regarded as un safe to human s and environment (Vinha , et

al., 2012; Nag , et al., 2013; Popoola, et al., 2013) .

In South Africa , 70 -80% of the rural population consults t raditi ona l healers for th e

most illness on a regular basis . This is mostly due to due to it accessibility and

affordability (Jager et al., 1996) . The combination of traditional and western medicine

is becoming increasing ly i mportant (Meissner, 2004). Medicinal p lants have curative

properties due to the presence of important bioac tive constituents (such as

phenolics, flavonoids, alkaloids, tannins, and terpenoids) known as secondary

metabolites that have a certain physiological action in the human body (Shaukat, e t

al., 2013).

The isolation of plant metabolites started in the nineteenth century. At the beginning,

structural elucidation of isolated compounds was limited by the technology available

at that time , it was done by classical and exhaustive degradative met hods and the

structures emanated from such were confirmed through synthesis then followed by

biological activity determinati ons.

Advancement of technology has made it possible for the structural classification of

chemical compounds to be done with only a few milligrams of pure material using

sophisticated instruments such as High -performance liquid chromatography (HPLC ),

Nuclear magnetic resonance (NMR ) and High -resolution mass spectrometry ( HRM S)

with many new compounds generated each year .

Some of these compounds are anti -malarial, anti -aging, anti -cancer , enzyme

inhibitors and some are known t o bind to specific receptors of pharmacological

interest. However, there are some compounds isolated through chemically guided

isolation that have prov ed to be bio logically active.

Problem statement

This study would investigate how to isolate some of the polar compounds in an

extracted fraction of Helichrysium cymsum . The compound isolated would be

identified using nu clear magnetic resonance .


The NMR can be used to isolate and identify polar co mpound s form plant extract

Helichrysium cy msum .

A ssumption

There would be more than one pure polar compound identif ied and would have

some medicinal properties.

D elimitation

Exploration of the isolated compoun d against any diseases. Conduct any

comparative phytochemical and biological analys es on the isolated chemical

cons tituents present in the Helichrysium species .

Literature Review

1. Asteraceae family

Asteraceae is the largest flowering family in the world, co ntaining 23600 species

allocated to 1620 genera and have an international distribution, but is especially

diverse in the tropical and subtropic al regions of southern Africa, the Mediterranean

region, central Asia, south -western China, and Aust ralia (Funk, et al., 2009).

Although the vegetation can vary, it can be recognised in which numerous small

flowers open first on the outside and only sometimes subtended by bracts. The

anthers are fused and form a tube through which the style extends befor e the two

stigmatic lobes separate and be come recurved, while the single -seeded fruit usually

have a plumose pappus (Galbany -casals, et al., 2014).

The Asteracea e family is divided into 13 subfamilies (Panero, et al., 2014) of which

the Asteroidea contains more than 70 % of the species currently recognised. Recent

molecular studies discovered thre e main lineages within the Asteroideae that have

been recognised at the super -tribe level (Robinson, 2004).

2. Helichrysium genus

Helichrysium is c lassified in the t ribe Gnaphalieae , which is known to be the largest

flowing plant . It currently contains 500 -600 species which are mainly found in Africa

and Madagascar, but also in Europe, Asi a and Australia (Hilliard, 1983; Manning &

Goldb att , 2012) . 240 -250 species occ ur in southern Africa of which approximately

70 -80 species are found in the uKhahlamba Drakensberg Mountains in KwaZulu –

Natal and in the Eastern Cape region in South Africa. Due to massive morphological

diversi ty the genus has been subdivided into 30 morph ologica l groups based on the

shape and the size of the flower heads (Hilliard 1983 ; Pooley 1998; Pooley 2003) .

South Africa Helichrysium species is associated with the treatment of infection of the

skin, such as circumcised wounds and wound dres sing (Loure ns, et al., 2008).

Helichrys ium species has proven to contain secondary metabolites , like phenolic, ?-

pyrone derivatives, and acetophenones derivatives (Lourens, et al., 2008), which

give s them global acceptability due to the large application in the cosmetic and

pharmaceutical fields acting as anti -infl ammatory (Viegas, et al., 2014) , antibacterial

and antioxidant agents (Kolayli, et al., 2010; Mari, et al., 2014; Rigano, et al., 2014).

Commercially drugs derived from this genus was used as a liver stimulant , diuretic .

It can act as an anti -inflammator y, tissue re -generating property and treatment for

some cardiovascular conditions (Kladar, et al., 2015).

3. Skin anti -ageing properties of Helichrysium species

Che mistry of metal -chelating ability of flavo noids with copper (II) in t yrosinase has

been studied with mild inhibitory activities demonstrated with compoun d H. Larix , H.

acucatum, H. kraussii, H. oreophilum, H. cymosu m & H. bracteatum . Other prior

activity of anti -tyrosinase data documented, were carried out using extract s obtained

from plant materials . However, details of the mechanism of chelation between

copper (II) and the chemical constituents of Helichrysium species remain

untraceable in the Sci Finde r databas e.

4. Antioxidant a ctivities of Helichrysium genus

The antioxidants activities of this ge nus were directed towards fraction obtained from

the crude plant materials . However, the compound responsible for this has only be

documented in a few cases ( Albayrak , et al., 2010 ).

A medicinal plant in the Mediterranean region was found to have good ant ioxidant

activity using th e2,2 -diphenyl -1-picryl hydra zyl (DPPH ) assay with the Ion

Chromatography 50 (IC 50) value 100 -fold higher than Trolox . It has a n adequate

amount of anti -aging prop erties due to its skin tissue regeneration ability and its

alleviation of inflammation through its anti oxidant properties, protecting the skin from

the damag ing effect of free radi cals ( Orn ano, et al., 2015) .

5. H. Cym osum sp . cymosum

The main species (fig 1) is a shrub that usually grows up to 1m tall standing erect

with thin greyish -white woolly branches densely covered wi th l eaves. The upper

surface of the leaf is covered in thin silvery grey, paper -like hairs . It has a bright

yellow canary in flat -topped flo werheads . Each flowerhead is a cluster of 6 -20

flowers. The flowers generally have smooth tips, and a pappus of many bri stles . H.

cymos um subsp. cymos um has been record ed from the Cape point upward through

the Eastern Cape and K ZN all along the coast.

Figure 1: A flowing shrub of H. cym sum sp. Cymsum

Three chemical constituents have been isolate d from H. cymo sum subs p. Cymosum .

Firstly, heli humulone (fig1 a), an oily phloroglucinol derivative with substantial

antimicrobial activity. The other specie isolated is 5 -hydroxy -8-methoxy -7-

prenyoxyflavanone (fig b). Lastly , a flavone derivative , helichr omanochalcone (fig c)

which was isolated from the roots ( Jakup ovic et al. 1989 ; V an Vuuren et al. 2006)

Figure 2: (a) Helihumulone, (b) 5 -hyd roxy -8-methoxy -7-preny oxyflavanone

and (c) a helichromanochalcone isolated f rom H. cymosum sp. cymosum

(Jakupovic et al. 1989; Van Vuuren et al 2006)

6. H. cymosum sp . calvum Hillard

This subspecies differs from H. cy mosum so cy mo sum on that it has a narrower leaf

and the branches are more densely clustered with its flowing hea d being smaller that

that of H. cymosum sp. cymosum . It can be differentiated by its glabrous ovaries . It

can be found in the Eastern Cape and K ZN. It only grows in areas that are higher

than 1200m above sea level up to a max imum of 3170m (Hillard 1983; Pooley 1998) .

This species i s known to contain several flavonoid derivative compounds and a few

terpene compounds (Bohlmann et al ., 1979) .

Figure 3: A flowing shrub of H. cymosum sp . clavum

Methodolog y

7. Plant collection

The Plant was collected at Hout bay Sea side Ca pe town South Africa. The study

would only focus on a n extracted fraction of the plant. It was prepared and stored at

Cape Peninsula University of T echnology (CPUT) .

8. Column chromatography

A sub -fraction of plant extract ed, was extracted by column chromato graphy using

ethanol (mobile phase) which was performed in sephadex as the stati onary phas e,

supported with glass co lumn of different sizes .

9. Thin layer chromatography (TLC)

Visualization of TLC plates was done by observing the bands “spots” after

developme nt under UV at ?254 nm and ?366 nm using UV lamp followed by dipping

into vanillin/sulphuric acid reagent . Chemical profiles of the fractions were identified

based on the colour produced after viewing under UV and then spraying with the

spray detecting rea gent (vanillin/sulphuric).

10. Material an d instrument

Extraction would take place at the CPUT laboratory with the use of col umn

chroma tography for extraction and nucle ar magnetic resonance (NMR) to identify the

isolated compound.

11. Schedu le

The project is estim ated to take a t least three month of laboratory work. The practical

aspect of it would be preformed at least once a week.

12. Budget

• Column chromatography

• TLC plate

• Et hanol

• Methanol

• Dich loromethane

• Acetone

• Rotary

• Vanillin

• Ultra -violet light



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