An overview of some socio-economic

aspects and scientific research on

MAPs in Albania

Prof. As. Vilma Papajani

University of Medicine

Tirana, Albania

MAPs of the Republic of Albania

--The Albanian flora contains 3250 plant species including aromatic and medicinal plants (MAPs more than 300 species). -The data show that this property has an 200 thousand tons overall capacity production (estimated at approx. $ 700 million)*. • -Albanians have traditionally used garlic, onion, chamomile, mountain tea, St. John’s worth, linden, sage, wild mint, mallow, chicory, juniper etc. as home remedies. *Source: THE MEDICINAL AND AROMATIC PLANTS VALUE CHAIN IN ALBANIA USAID/ALBANIA AGRICULTURE COMPETITIVENESS (AAC) 2009 •

INDIVIDUAL

Profile of main operators in the MAPs value chain of Albania

Source: THE MEDICINAL AND AROMATIC PLANTS VALUE CHAIN IN ALBANIA

USAID/ALBANIA AGRICULTURE COMPETITIVENESS (AAC) 2009

The MAPs generate more than 16 m Euro per year. MAPs bussines includes about 100.000 persons.

COLLECTORS

COLLECTORS

REGIONAL REGIONAL

HARVESTERS, PRODUCERS OF

CULTIVATED MAPs

HARVESTERS, PRODUCERS OF

CULTIVATED MAPs

PROCESSORS AND EXPORTERS Small processors Medium and large processors

PROCESSORS AND EXPORTERS Small processors Medium and large processors

Collection of MAPs

Collectors

Individual collectors Regional collectors

In each region there are at least 2-4 main collectors. Aprox. 20-100 tons of MAPs annually. Malesia Madhe is an exception with around 6-10 collectors.*

(Number: 30)

** ***

Source: * THE MEDICINAL AND AROMATIC PLANTS VALUE CHAIN IN ALBANIA USAID/ALBANIA AGRICULTURE COMPETITIVENESS (AAC) 2009 **Published by Academy of Sciences. *** Published by Ministry of Environment, Forests and Water Administration

PROCESSORS AND EXPORTERS 9-10 medium and large processors -trade between to 500-3000 tons of MAPs and WGPs

annually, divided into: semi ready products according to the customers demand

for dimensions and packaging and essential oils (produced 35 and 40 tons annually)*

Source:* THE MEDICINAL AND AROMATIC PLANTS VALUE CHAIN IN ALBANIA USAID/ALBANIA AGRICULTURE COMPETITIVENESS (AAC) 2009,

‘**The Organic Standard', www.organicstandard.com

National and foreign certification bodies operate in Albania. BCS, ICEA ,CERES, SKAL, bio.inspecta and Italian Codex have certified exporters of medicinal plants and essential oils.**

Only few companies produce herbal medicinal teas and traditional herbal medicines such as syrupus, oils etc. A lot of these preparations come from imports (Europe, USA etc.)

Manufactures of MAPs

Export of MAPs and other WGPs from Albania

Source: UNSTAT

Albania is exporting a wide range of products, but is particularly strong in sage, oregano, thyme , savory and last years in wild apples. Sage accounts: 2,000 to 2,500 Mt per year. Germany and USA are the main markets (about 60% of total Albanian MAPs exports). Other important markets are France, Turkey and Italy. Exports have increased over the years; in 2008 recorded over 8367 tons and in 2011 recorded 8780 tons (with a value of $ 23 million) Source: UNSTAT

13.827

12.5

40

32.4

25

18.2

50

46.240.6

62.5

81.8

50

15-29

YEARS

30-44

YEARS

45-59

YEARS

60-74

YEARS

>74

YEARS

>10 TIMES IN A YEAR

5-10 TIMES IN A YEAR

<5 TIMES IN A YEAR

FREQUENCY OF USE OF MEDICINAL PLANTS

ACCORDING TO AGE

MEDICINAL AND AROMATIC PLANTS MAINLY USED IN THERAPY

Pasho I. Papajani V., “Study of the aplication of Phytopherapy in Albania (Tirana area)”, Conference of Bio-Medical Sciences, 2013, Tirana, Albania.

The use of medicinal and aromatic plants

Research on Medicinal and Aromatic plants in Albania

The studies of MAPs in Albania have been and are

subjects of scientific research units as:

Academy of Sciences, ex-Institute of Biological

Research

Dep. of Chemistry, Dep. of Biotechnology Faculty of Natural Sciences, University of

Tirana.

Department of Pharmacy, session of Pharmacognosy, Faculty of Medicine,

University of Tirana (Nowdays University of Medicine)

The Traditional Medicine Institute

(1956-1992)

Other institutions also deal with evaluation of chemical composition of MAPs as Agricultural University of Tirana etc.

Research on MAPs The object of research are the most economically important MAPs such as: • Salvia officinalis L. • Thymus spp. (Thymus vulgaris L., Thymus longicaulis C. Preisl. , Thymus capitatus Hof. et Link) • Origanum spp. (O. vulgarae L., O. heracleoticum L syn. O. vulgare sub. hirtum ) • Saturea spp. • Mentha spp. • Hypericum perforatum L. • Crataegus spp. • Tilia spp. • Sambuci spp. • Sideritis raeseri Boiss. & Heldr. • Silybum marianum (L.) Gaertner

Main lines of Research on MAPs Pharmacognosy group, Faculty of Pharmacy

• Evaluation of MAPs wild growing in Albania. - Chemical characterization of their essential oils as a contribution in

compilation of ‘the map of chemotypes’ in the service of the collectors, cultivators, exporters of aromatic-medicinal species.

- The dynamics of accumulation of active compounds in medicinal plants.

- Monitoring of seasonal variations and correlation among biochemically related compounds. (Th. vulgaris, O. heracleoticum).

0

0.5

1

1.5

2

2.5

March April May JunePe

rce

nta

ge o

f e

sse

nti

al o

ils

Seasonal variations of thyme essential oil yield.

0

10

20

30

40

50

60

70

80

March AprilMay June

The

co

nce

ntr

atio

n o

f m

ain

co

mp

ou

nd

s

The variability of phenols and terpenes during vegetative stages

r=-0.99, p<0.001

Phenols

p-cymene and γ-terpinene

Source: Papajani V., Hatipi M., Troja E., Haloci E., “The correlation among phenolic monoterpenes and their precursors in essential oil of Th. vulgaris L.” ; The Natural Sciences Bulletin, 2012, Tirana, ISSN: 224-1779, p.168-174. *

*

Chemical characterization of MAPs essential oils

-The oils of white and red oregano were found to be rich in carvacrol

-Two chemotypes of white oregano: carvacrol/carvacrol and thymol

-Three chemotypes of Salvia officinalis were detected.

-Rosmarin oils were more similar to those of Italian origin due

to their 1.8 cineole, camphor, α-pinene contents.

Chemical composition of red oregano

The content (%) of carvacrol and thymol in white oregano essential oils from areas of Albania.

Research on MAPs

Publication from 2003- • Asllani U., Papajani-Toska V. (2003) “Chemical Composition of Albanian Thyme Oil.”; published in the "Journal of Essential oil Research “, 15,

page. 165-167, May/June 2003, USA. • Asllani U. “Essential oils of medicinal aromatic plants from Albania”, published by ILAR, Tirana, Albania 2004. • E. Kongjika, A.Bacu, V. Papajani etc. "Scientific assessment of some aromatic medicinal plants, their cultivation perspective", coauthor, Tirana

2005, ISBN 99943-656-7-3. • Asllani U., Papajani-Toska V. (2001) "The chemistry of phenolic essences in some aromatic plants in Albania", in the framework of the Italian-

Albanian cooperation (Interreg II) on valuation of biodiversity. 4-th seminar, Tirana. Material published on the project summary. • Papajani-Toska V. " Origanum heracleoticum L., “The chemistry of essential oils from plants of our country." II-nd Pharmaceutical Scientific

Conference , Tirana, April 2001 • Papajani-Toska V., Koci K. (2004) "Changes in chemical composition according the essential oil phenophases of Origanum vulgare var. viridis

cultivated in the Ishëm - Manëz area.", published in "Biological Studies" (edition of the Academy of Sciences and the Biological Research Institute), No. 8, Tirana, p. 9-14.

• Babani F., Koci K., Papajani-Toska V. (2005) "Chemotype and genetic characterization of four Albanian populations of Salvia officinalis L." published in "Biological Studies" (edition of the Academy of Sciences and the Biological Research Institute), No. 10, Tirana p.69-77.

• Papajani-Toska V., Asllani U. (2005) "Chemical composition of the essential oils in Corydothymus capitatus L. Reiche " Scientific bulletin of " Aleksandër Xhuvani" University UNIEL, Natural Sciences series, No. 2, Elbasan p.47-53.

• Babani F., Toska V. Koci K., Misiri E. (2006) "Characterization of essential oil components in some aromatic medicinal plants." Workshop: "Scientific Assessment of some aromatic medicinal plants. Perspective of their development", in the framework of the National Programme for research and development "Biotechnology and Biological Diversity" organized by the Ministry of Education, Academy of Sciences. Tirana, January 2006.

• Lazari D., Mpompou E.-M., Papajani-Toska V., Kontogiorgis C., Hadjipavlou-Litina D., Chemical Composition and Antioxidant Activity of the Essential Oils isolated from Greek and Albanian Thymus species. 13th Hellenic Symposium on Medicinal Chemistry (HSMC-13), 14-15 March 2008, Athens, Greece.

• Myftari B., Papajani V., Asllani U. (2009) “Comparative study on essential oil of Salvia officinalis L. from different areas of Albania”; Skin and formulation 3-rd symposium of skin Forum 10th annual Meeting, Versailles, France, 9-10 March 2009.

• Toska-Papajani V., Haloci E. (2009) “ Identification and quantitative determination of chemical components of Hypericum perforatum L. and its pharmaceutical forms”; annual meeting of Alb Shkenca Institute: september 2009, Tetovo, Macedonia.

• Papajani V., Asllani U., Troja E. (2009)“ Chemical composition of Albanian Wild oreganos oils”; World Congress of pharmacy and Pharmaceutical Sciences, 69th international congress of FIP, 3-8 September , Istanbul, Turkey, Book of Abstract..

• Papajani V., Troja E., Miraci M., Asllani U. (2010) “Chemical composition of the essential oils from two wild species of Thymus growing in Albania.”, World Congress of Pharmacy and Pharmaceutical Sciences,70th International Congress of FIP, 09/2010, Lisbone, Portugaly, Book of Abstract.

• Papajani V., Asllani U., Haloçi E. (2010) “ Essential oils variability in Saturea montana L. from Albania”; World Congress of Pharmacy and Pharmaceutical Sciences: 70th International Congress of FIP, 09/2010, Lisbone, Portugaly, in Proceeding.

• Papajani-Toska V., Hatibi M., Basha B., Asllani U. (2011) “Essential oils composition variability in Salvia officinalis L. from Albania and Kosova”; Fifth Congress of Pharmacy of Macedonia with International participations, Ohrid Macedonia, in Proceedings.

• Papajani V., Dylgjeri K., Troja E., Haloci E. (2011) “Chemical composition of Rosemary oils from Albania”; Fifth Congress of Pharmacy of Macedonia with International participations, Ohrid Macedonia, in Proceedings.

• Papajani V., Hatipi M., Troja E., Haloci E., “The correlation among phenolic monoterpenes and their precursors in essential oil of Th. vulgaris L.” ; The Natural Sciences Bulletin, 2012, Tirana, ISSN: 224-1779, p.168-174.

Projects supported by international and national grants.

• 2001-2002 International project : “The Italian and Albanian cooperation, for a contribution to a better assessment of biodiversity”, with the participaotin of Mediterranean Agronomic Institute of Bari, University of Bari, University of Lecce, Albanian Ministry of Environment, Tirana University Department of Pharmacy, Institute of Biological Studies.

• 2003-2005 National project: “Evaluation of the potential capacity, biodiversity identification on molecular level, propagation in “vivo” and “in vitro” for the cultivation of optimal chemotypes of some aromatic-medicinal species in poor soils of Albania”, in the framework of The National Research and Development Programe "Biotechnology and diversity ", with the participation of the Biological Research Institute, University of Tirana Department of Pharmacy, Forest and Pasture Research Institute.

• 2005-2007 International project: “Exploitation of pharmaceutical species with economic interest of genus Thymus (Thyme)”, with the participation of Laboratory of Pharmacognosy Aristotle University of Thessaloniki (AUTH) and Department of Pharmacy Tirana University, Faculty of Agronomy of the Agricultural University in Tirana.

• 2005-2007 International project: "The use of combined technologies, packaging in modified atmosphere, vacuum packaging. Essential oils for the preservation and safety of poultry for consumption”., with the participation of the University of Ioannina, Greece, Tirana Faculty of Natural Sciences and Department of Pharmacy.

• 2008-2010 International project : "Study of the effect of several new molecules on macrophage phagocytic ability”, with the participation of the Department of Experimental Medicine and Biochemical Sciences, University Tor Vergata, Rome and Department of Pharmacy, University of Tirana.

• 2010-2012 SEE-ERA.NET Plus Joint Call “Conservation and utilization of the diversity of sage species (Salvia spp.) traditional food preservatives and spices SEE-ERA.NET PLUS Institute of Agrobiotechnology, Greece; Dpt. of Biotechnology, Univ. of Tirana; Inst. of Pharmacognosy, University Ss Cyril and Methodius, Macedonia; University of Zagreb, Faculty of Agriculture; Institute of Botany and Pharmacognosy, University of Wien .

• 2012-2014: International project . "Extraction and characterization of substances containing sulfur / selen in aromatic and officinal plants with antitumoral potential action." With the participation of the Department of Pharmacy, University of Tirana and Department of Sciences and Chemical Technologies , University Tor-Vergata, Rome, Italy.

CO2 extraction (high pressure methodology ) applicated in our science research activities

(Chemistry group in collaboration with pharmacognosy group )

Application of modern methods for extraction of bioactive compounds from MAPs

Operating condition

Pressure values = 55 - 60 bar

Temperature range = 308 - 312 K

Plant material= 10 - 20 g

Diameter of granules = 0.8 - 1.2 mm

Quantity of CO2 required = 220 ± 10 g

Evaluation of essential oils from different extraction techniques

• Papajani V., Troja E., Çomo A. (2008) “The use of the effective methods for the extraction of the essential oils from Albania medicinal plants”; International Conference on Biological and Environmental Sciences, Tirana, 26-28 September 2008. Proceeding p. 90-95.

• Papajani V., Troja E., Como A.; (2008) "Efficiency of CO2 in extraction of essences from Albanian medicinal plants", International Conference on Biological and Environmental Sciences, 26-28 September, Faculty of Natural Sciences, Tirana, Albania, Proceedings, ISBN : 978-99956-05-85-8, f.90-95.

• S. Abazi, O. Xhika, H. Fico; “Extration of Hypericum perforatum L. using hexan, Dcm and liquid CO2” , In. Journal of Ecosystems , Volume 1/1, 2011, pg. 63-68.

• S. Mance, S. Abazi; “Extraction of Eugenol from Eugenia carophyllata with different extraction methods”, International Conference of Ecosystems (ICE) 2012, Tirana, Albania.

0.00% 0.50% 1.00% 1.50% 2.00%

Th vulgaris

Th. longicaulis

R. officinalis

0.28%

0.50%

0.43%

0.79%

0.40%

1.79%

Yield %

Aro

ma

tic

pla

nts

Comparison of yield values between subcritical CO2 and hydrodistillation

• CO2 subcritical

• Classic method

• Antimicrobial activity (in vitro) (in collaboration with food chemistry group)

• Antimicrobial effects of some essential oils from:

Assessment of biological/pharmacological activity

Bacteria Bacillus gram+, S. aureus Escherichia coli, Proteus vulgaris Yeasts Saccharomyces spp. Rhodotorula spp. Moulds Aspergillus niger Penicilium sp. Aspergilus flavus

1. Thymus vulgaris L. (thyme) 2. Thymus longicaulis C Preisl. (creeping thyme) 3. Rosmarinus officinalis L. (rosemary) 4. Saturea montana L. (savory)

Evaluation of antimicrobial activity

Media : MA, Capek DOX agar, Sabouraud, PDA, PCA

Determination of

the optimal media most sensitive microorganism effectivity of essential oils minimal inhibitory concentration –MIC

Metodology : Kirby Bauer Disk Diffusion Test

Evaluation of antimicrobial activity Application of these antibacterial activity has been evaluated and finds

applications in some fields like :

- Cereals (antimycotic - aspergillus spp.)

- Vegetables stored by freezing processes,

-Poultry meats & skin (chickens for consume, turkeys)

- Internal animal organs for consume (spleen, liver)

- Red muscle meat (replacement of antibiotics)

Inhibition zone M1

Inhibition zone M2

Inhibition zone M3

Inhibition zone M4

Inhibition zone M5

10 ug sample

24.8 25.5 25.2 26.1 27.5

Evaluation of antimicrobial activity Inhibition z.M1

Inhibition z.M2

Inhibition z.M3

Inhibition z.M4

Inhibition z.M5

Inh. z.1 Inh.z.2 Inh.z.3 Inh. z.4 Inh.z.5

30 32 29 29 29

23 30 29 27 28

28 28 28 30 31

0 20 40 60 80 100

10 µg

22 24 26 28

10ug sample

Satureja montana essential oils inhibition zone / S.Aureus

Inhibition z.1

Inhibition z.2

Inhibition z.3

Inhibition z.4

Inhibition z.5

Biodiversity of wild and cultivated populations of

aromatic-medicinal species via molecular markers (Biotechnology group)

• Application of molecular tools to study genetic polymorphisms at MAP, evaluation of suitable genotypes and effective use of

genetic resources.

Populations of Origanum vulgare sub. hirtum

1. Llogora, 2. Dajt, 3. Gjirokaster, analysed

by RAPD-randomly amplified polymorphic

DNA ( from right to left).

Cultivated and natural populations.

Three populations of Salvia officinalis (Llogora-1,

Gjirokaster-2, Dajt-3, analysed by markers SSRs-simple

sequence repeats).

AFLP (Amplified Fragment Length Polymorphism)

Analysis of the aromatic medicinal species S. officinalis and O. vulgare sub. hirtum

Publications: • Bacu-Grazhdani A., Kongjika E., Mullaj A.; Use of RAPD and SSR for

the identification of polimorphism among ecotypes of Salvia officinalis and Salvia triloba grown in Albania; Acta Physiologiae Plantarum, Vol.26, No 3 and poster presentation at the FESPB Congress held in Cracow, Poland,2004.

• Grazhdani-Bacu A., Makris.A, Owen. C, “ Expression patterns of two unique MTP gene fragments isolated from Salvia officinalis and Salvia fruticosa”, Biological Studies 8/2004:3-8

• Ariola Bacu, Alfred Mullaj, Fatbardha Babani, Efigjeni Kongjika. Sherbela (Salvia officinalis) of Albania: Evaluation of interpopulations variability via molecular methods.Second Conference of Institute Alb-Shkenca, Prishtina-2007. Poster.

• Kongjika E., Bacu A., Babani F., Zekaj Zh. “Biodiversity of Albanian populations of some aromatic medicinal plants in molecular, biochemical, histological level and some alternatives of ex situ conservation”. Italus Hortus, 2008. Proceedings of the VIII National Congress “ La Biodiversita-una risorsa per sistemi multifunzionali”, Salerno, Italy.

• A. Bacu, V. Mata, E. Kongjika, F. Babani, L. Dinga, F. Bakiri, 2008, “Micropropagation and “in vitro” conservation of the germplasm of some aromatic-medicinal species”, Proceedings of “International Conference on Biological and Environmental Sciences”, Tirana, Albania.

• A.Bacu, A. Loeser, C. Appenroth; “Amplified Fragment Length Polymorphisms (AFLPs) group populations of Salvia officinalis of Albania in accordance to their geographical origin”; International conference of Alb-Shkenca, Macedonia, 2009.

“IN VITRO” CULTIVATION AND THE ESTABLISHMENT OF THE GENETIC BANK FOR THE AROMATIC MEDICINAL SPECIES

Establishment of the genetic bank for some MAPs species

.

Genetic Bank for the two populations of Origanum vulgare L.

“In vitro” cultivation of S.officinalis . S. montana L.

The quality control and standardization of herbal drug raw material

Determination of heavy metals on MAPs and medicinal plants used as herbal tea

Publications 2005-2012

• Papajani-Toska V., Lazo P., Prifti I. (2008); "Determination of heavy metals in some endemic medicinal plants" Albanian Journal of Pharmaceutical Sciences, no. 1, June, p. 11-16.

• Papajani-Toska V., Sima Z., Vito S. (2005) "Considerations on the use and good- marketing of herbal medicines in Albania", Scientific Conference of Albania Pharmacists Association.

• Papajani V., Lazo P., Prifti I. “Determination of heavy metals in some Albanian medicinal plants used as herbal tea”; World Congress of pharmacy and Pharmaceutical Sciences, 69th international congress of FIP, 3-8 September 2009, Istanbul, Turkey, Book of Abstract.

• Papajani V., Vito S., Troja E., Miraci M, (2009) "Some aspects of quality control and safety of medicinal plants and their preparations," - Quality infrastructure ISBN: 987-99956-772-0-6, p. 167-172.

• Papajani V., Lazo P., Lubonja J., Shengjergji D., (2012) “Determination of heavy metals in some Albanian aromatic medicinal plants”; The Natural Sciences Bulletin, Tirana, ISSN: 224-1779, p.431-438.

Recent trends and future prospective • Extraction and characterization of sulfur/selen compounds in

aromatic and officinal plants with antitumoral potential action (mainly Allium L. species).

International project (2012-2014) cooperation between Albania and Italy (Pharmacognosy group, University of Medicine, Tirana and Biotechnology group, University Tor-Vergata, Rome).

• During the first year (2012-2013): Soluble OSC-conjugates were obtained by a new extraction protocol of OSCs from garlic in the presence of GSH at low temperatures.

• In process: The evaluation of water soluble OSCs-conjugates to inhibit the TST activity. This study opens the way to the identification of novel OSCs-conjugates obtained by a new

extraction protocol and highlights on their antitumor properties for therapeutic applications.

GC/MS LC-MS

• Chemical characterization and assesment of antioxidant activity in MAPs with polyphenols:

S. raesery, V. agnus castus (Fructus et Folium),

Origanum spp., R. officinalis, etc.

• Chemical and pharmacological evaluation of extracts from MAPs using CO2 extraction method.

• Ethnomedicinal and ethnobotanical studies in Albania etc.

Recent trends and future prospective

Thank you