YOUNG SCIENTIST PROGRAM

SMK (P) METHODIST,KUANTANPAHANG

Zingiberaceae

1.0 INTRODUCTION

ORGANIZERS :

KULLIYYAH OF PHARMACY,UIAM

&JPN PAHANG

1.2 Objectives of the program:

To create awareness on the importance of biodiversity as a source of medicine and food supplement

To educate the importance of biodiversity protection and conservation

To promote interest in scientific research

1.3 TASKS

• 1.3.1 Herbal Phytochemical Extraction and Screening

• 1.3.2 Jungle Exploration and Botanical Collection of Medicinal Plants

1.4 Objectives of the Tasks

• To screen the existence of alkaloid and flavonoid from the 2 specimens of Zingiberaceae family (Zingiber sp. & Elettariopsis sp.) collected at Taman Pertanian.

• To preserve and deposit the studied medicinal plants as herbarium specimens

2.0 Introduction to Zingiberaceae family

2.1 General characteristics

Consist of about 47 genera, 1400 species Perennial tropical herbs, ground flora of lowland

forest (Purseglove, 1975) Aromatic leaves, fleshy rhizomes, elliptic to elliptic-

oblong leaves arranged in two rows on the leaf shoot (Habsah et al.,2000)

Zingiber=bull’s horn (Sanskrit) (Larsen et al., 1999)

Cardamom(Elettaria cardamomum)

LempoyangZingiber zerumbet

Common gingerZingiber officinale

Turmeric(Curcuma domestica)

Galangal (Alpinia galanga)

Elettariopsis sp.

• ‘Cawak’• Herbaceous,

underground rhizomes, 1m

• Dark green, thick, soft leaves

• Leaves sprout out from the rhizomes

• Leaves curve downwards in one direction

• Aromatic leaves

2.2 HabitatGround plants of tropical forestDamp, humid, shady

2.3 DistributionIndia, tropical Asia, northern Australia (Tuntiwachwuttikul et al.,1986)Indonesia, Malaysia, Singapore, Brunei, Philippines, Papua New Guinea (Puangpen, 1999)

2.4 Classification4 tribes-Hedychieae-Zingibereae-Alpineae-Globbeae

3.0 Chemistry• Rich in terpenoids• Alkaloids detected (Larsen et al.,1999)• Phytochemical analysis of the methanol extract-presence of

alkaloids, glycosides, tannins, flavonoids, sterol, triterpenes and oils

• Essential oils cause the fragrance• Main constituents-sesquiterpenoids with zingiberence as main

component• Pungent taste-due to nonvolatile phenylpropanoid-derived

compounds particularly oleoresins known as zingerone, gingerols & shogaols

• Gingerols increase motility of gastrointestinal track, have analgesic, sedative, antipyretic & antibacterial properties

• Gingerols can kill ovarian cancer cells (University of Michigan)

4.0 Uses

• 4.1 Culinary Use-Young ginger rhizomes-pickled in vinegar or

sherry (best eaten with sushi)-Ingredient in dishes (flavouring)-curry,stew etc-Ginger tea

4.2 Medicinal Use Chinese-dried ginger to supress cough, colds, flu.

Dr. Halijah Ibrahim :`Cultivated Gingers of Peninsular Malaysia: Utilization, Profiles and Micropropagation’-anti-inflammatory properties, opens up blood vessels, creates sweat & warmth, good for arthritis, nausea & vomiting and helps with motion sickness.

In the West - dried ginger root powder (sold in capsules)

FDA’s generally safe list.

Ginger may combat deadly infant diarrhea in developing world-Science Daily (Oct 4, 2007)

Research on rats suggests ginger as a useful treatment for diabetes (Somchit et al, 2005).

Curcuma zedoaria (Zedoary Lily)-ulcers, wounds, tumours, artherosclerosis, inflammation-Ohyul syndrome-blood stagnation (Seo et al.,2005)

Zingiber zerumbet (Lempoyang)-inflammation-Zerumbone-chemopreventive properties (Jang et al.,2005)

Elettariopsis curtisii Baker-essential oil –antioxidant & antibacterial (Vanida et al.,2008)

4.3 Beauty Tips

• Antioxidant properties- slow down aging process

• Plant’s fresh juice-applied to the skin to treat burns.

• Essential oil- pain relief

5.0 ACTIVITIES5.1 SESSION 1 : JUNGLE EXPLORATION & BOTANICAL COLLECTION OF MEDICINAL PLANTSDATE : 6 FEB 2010 (SAT)VENUE : TAMAN PERTANIAN

• 4 Zingiberaceae specimens collected including rhizomes. In duplicate.

• Cleaned off soil. Labeled & photographed• Pressed with old newspaper.• Methanol-stop enzymatic activities• 2 specimens-phytochemical extraction & screening.

Dr. Wan Azizi leading the expedition. Students (eager beavers) listening

attentively to the briefing.

Collecting our first Zingiberaceae sample. Dig Shy dig!

Another Zingiberaceae sample.

Biodiversity witnessed during the expedition.

Girls, what tree is this?

Sure budak-budak bandar

ni tak tahu!

Specimens were cleaned and then carefully pressed with old newspaper.

Dr. Norazian demonstrating how to prepare the herbarium

specimens.

This frame will ensure that all the

herbal specimens are evenly pressed!

This wooden frame is

imported from

Germany!Expensive

you know? !

Then place one frame on top and

one at the bottom of the

stacked specimens.

5.2 SESSION 2 : HERBAL PHYTOCHEMICAL EXTRACTION AND SCREENINGDATE : 7 FEB 2010 (SUN)VENUE : KULLIYYAH OF PHARMACY, UIAM

Alright, what are phytochemicals?

? ? ??

SCREENING OF ALKALOID & FLAVONOID FROM PLANT SAMPLES

• 1.0 Objectives

(i) To prepare plant extract(ii) To screen the existence of alkaloid

& flavonoid from plant samples

2.0 Introduction to Phytochemicals

• Chemicals that are found in plants, that may have beneficial effects on our health.

• Exist naturally as secondary metabolites• E.g. phenolic compounds - flavonoids, terpenoids, alkaloids

Alkaloids

- Nitrogenous organic molecule that has pharmacological effects on humans. - Name derived from `alkaline’. - Originally the term was used to describe nitrogen-containing base (an amine)- Are found in plants, animals & fungi. Can be

extracted with acid (HCl, H2SO4, maleic acid & citric acid)

Flavonoid

- Collectively known as vitamin P & Citrin- Organic pigments in plants- A natural antioxidant- Helps maintain body’s health & protect against diseases. - Protects the membranes from being damaged, prevents hardening & furring up of arteries.- Inhibits growth of bacteria & viruses.

3.0 Materials & Apparatus

Herbal samples (Zingiber sp. and Elettariopsis sp.)Conical flaskBeakerAcid washed sandSeparating funnelEthanolTest tubeMayer’s ReagentMortar

Scientific classification:Kingdom : PlantaeOrder : ZingiberalesFamily : ZingiberaceaeGenus : ZingiberSpecies : Unknown

Scientific classification:Kingdom : PlantaeOrder : ZingiberalesFamily : ZingiberaceaeGenus : ElettariopsisSpecies : Unknown

3.0 MethodsA. Preparation of plant extracta. 20 gram of fresh materials were cut into small pieces

and put in the mortar. 1 spatula of acid washed sand was put into grind samples

b. The sample was transferred into a small beaker or conical flask and 30 ml of ethanol was added.c. The samples were boiled for 10-15 minutes.d. The mixture was poured into separating funnel line

with filter paper, into a conical flask.e. The flask and the plant materials were rinsed with ethanol then filtered.f. The flask was put in the steam bath and the extract was concentrated to about 20 ml.

B. Alkaloid Test

• a. 20 gram of fresh materials were cut into small pieces and put in the mortar. 1 spatula of acid washed sand was put in to grind samples.

• b. The sample was transferred into a conical flask and 20 ml of ammoniacal chloroform was added and filtered.

• c. The filtrate was transferred into a test tube and 15 drops of aqueous sulphuric acid 0.5M added.

• d. Mixture was shaken and let to settle to form 2 layers.• e. The aqueous layer (upper layer) was separated and

transferred into a test tube.• f. A few drops of Mayer’s Reagent were added. Creamish

precipitate indicated the presence of alkaloids. Result was observed and recorded.

C. Flavonoid Test

• a. 10 g of plant materials were boiled with 20 ml distilled water on the water bath.

• b. Extract was filtered.• c. 5 ml of the filtrate was transferred into a test tube

and concentrated to dryness on the boiling waterbath.

• d. The residue was treated with 0.1M NaOH, and 0.1M HCl was added.

• e. Solubility and colour changes were observed. A yellow solution with NaOH, which turned colourless with dilute HCl confirmed the presence of flavonoids.

Clean the rhizomes &

leaves before you cut & grind

them!

I’m grinding the macerated

leaves!No, I’m not

playing masak-masak!

I need 20 ml of distilled water!

I’m pouring the mixture while hot, through separating funnel lined with

filter paper.

Concentrate the extract to about

20 ml.

Add a few drops of Mayer’s Reagent. Presence of

creamish precipitate indicates the presence of alkaloid!

Yes! Yes?

4.0 Results

I. Phytochemical screening

Specimen/Test for

Zingiber sp Elettariopsis sp

Leaves Rhizomes Leaves Rhizomes

Alkaloid Creamish precipitate

present

Creamish precipitate

present

Creamish precipitate

did not present

Creamish precipitate

did not present

Flavonoid Solution did not turn

colourless with dilute

HCl

Solution did not turn

colourless with dilute

HCl

Solution turned

colourless with dilute

HCl

Solution turned

colourless with dilute

HCl

6.0 Discussion

• Zingiber sp. was found to contain alkaloid in the leaves and rhizomes.

• Elettariopsis sp. was found to contain flavanoid in the leaves and rhizomes.

• The results are consistent with most studies that showed Zingiberaceae (Ginger family) contain phytochemicals (eg alkaloids, flavonoid, carotenoids, oleresins like gingerol and shogaol).

• Therefore these two species have high potential of being developed and cultivated for their medicinal and other uses.

THE STAR (27 FEB 2010)

7.0 Conclusion

• Zingiber sp contained alkaloid and did not contain flavonoid.

• Elettariopsis sp. contained flavonoid and did not contain alkaloid.

II. Herbarium specimens

Zingiber sp.(Ginger family)

Elettariopsis sp.(Cawak)

Scaphoclamys sp.(Kunyit Landak)

Etlingera sphaerocephala(Kantan)

8.0 ReferencesInternet websites:• Wikipedia org/wiki/Zingiberaceae• Johor Biodiversiti Database http://www.iwana.com.my/jbiotech/page.php?pageid=s_id=22a&search1 =Elettariopsis%20sp• http://www.botany.hawaii.edu/herbarium/digitallib/collections/hlaplants/pdfs/Zin_Elettariop sis_z

zsp_L990478_w.pdf• Antiooxidant properties of some exclusive species of Zingeberaceae Family of Manipur• http://ejaafche.uvigo.es/index.php?option=com docman&task=doc view&gid=117• http://www.informaworld.com/smpp/content~content=a904291009&db=all• http://eprints.usm.my/8891/1/CLONAL PROPAGATION OF Curcuma zedoria ROSC.pdf• Zinger zerumbet (LEMPOYANG) A POTENTIAL ANTI-INFLAMMATORY AGENT• http://pkukmweb.ukm.my/~rsenr1/P516-520.pdf• Screening of Zingiberaceae extract for antimicrobial linkinghub.elsevier.com/retriever/pii/so3788741000002233• Angiosperm Families Zingiberaceae delta-intkey.com/angio/www/zingiber.htm• Phytochemical with hypoglycemic potential linkinghub.elsevier.com/retrieve/pii/so378874106001656• Antipyretic & Analgesic activities of Zingiber pkukmweb ukm.my/-resin r/3/sen11/p692-697 pdf• Pharmacological basis for the medicinal use of ginger. cat.inist fr/?amodele=amodele=affiche

9.0 Acknowledgements

• Education officers of JPN Pahang• Lecturers, staff & students of Kulliyyah of

Pharmacy, UIAM• MGS - Principal, teacher advisors & friends

The roots of education

are bitter but

the fruit is

sweet!-Aristotle