WORLDWIDE PHARMACEUTICAL FIELD

WESTERN COUNTRIES

DEVELOPING COUNTRIES

PERCENTAGE OF DRUGS OF BOTANICAL ORIGIN

20

80Data from WHO (2002)

PlantPapaver somniferum L.

Strychnos nux - vomica L.Cephaelis ipecacuanha (Brot.) TussacCinchona ledgeriana Bern. Moens ex Trimen

Coffea arabica L.Colchicum autumnale L.Filipendula ulmaria (L.) Maxim.Atropa belladonna L.Theobroma cacao L.Erythroxylum coca Lam.Physostigma venenosum Bal.Pilocarpus jaborandi HolmesDatura metel L.Hyoscyamus niger L.Ephedra sinica Stapf.Digitalis purpurea L.Rauwolfia serpentina L.

Chondrodendron tomentosum Ruitz et PavonCatharantus roseus (L.) G. Don

Ammi visnaga (L.) Lam.Silybum marianum (L.) Gartn.Coleus forskohlii Brig.Taxus baccata L.Camptotheca acuminata L.

AgentMorphineNoscapineCodeinePapaverineStrychnineEmetineQuinineQuinidineCaffeineColchicineSalicinAtropineTheobromineCocainePhysostigminePilocarpineScopolamineHyoscyamineL-EphedrineDigoxinAjmalineReserpineRescinnamineTubocurarineVinblastineVincristineVisnadineSilybinForskolinPaclitaxelCamptothecin

ActivityNarcotic analgesicAntitussiveAntitussive, narcotic analgesicSmooth muscle relaxantCNS stimulantAmebicideAntimalarialAntiarrhytmicCNS stimulantAntinflammatory (gout)AnalgesicAnticholinergic, mydriaticSmooth muscle relaxantTopical anestheticCholinergicAntiglaucoma, mioticAnticholinergicAnticholinergicSympathomimeticCardiotonicAntiarrhytmicAntihypertensiveAntihypertensiveSkeletal muscle relaxantAntitumorAntitumorCoronary vasodilatorAntitoxic, liver protectantAdenylate cyclase stimulatorAntitumorAntitumor

Year of Isolation1806181718321848181718171819183318191820182918311842186018641875188118811897193019311952195419351952195819611968197719911993

PLANT DERIVED ACTIVE AGENTS

PlantAgave sisalana (Engelm.) J.R. Drumm et Prain

Dioscorea sp.

Glycine max. (L.) Merr. (Soya bean)

Colchicum autumnale L.

Gloriosa superba L.

Podophyllum peltatum L.

Solanum sp.

Catharantus roseus (L.) G. Don (aerial parts)

Catharantus roseus (L.) G. Don (roots)

Synthone Agent

Ipomoea hederacea Jacq.

hecogenin

diosgenin

stigmasterol

colchicine

colchicine

podophyllotoxin

solasodine

anhydrovinblastine

serpentine

lysergol

corticosteroids

corticosteroids, sex hormones

thiocolchicoside

thiocolchicoside

etoposide, teniposide

Taxus baccata L. (leaves) 10-deacetylbaccatin III taxol, taxotère

navelbine

raubasine

nicergoline, pergolide

Voacanga africana Stapf. tabersonine vincamine, vinpocetine

corticosteroids, sex hormones

corticosteroids, sex hormones

Camptotheca acuminata L. camptothecin topotecan, irinotecan

ACTIVE AGENTS FROM PLANT DERIVED SYNTHONES

Taxus baccata L. (leaves) 10-deacetylbaccatin III taxol, taxotère

10-deacetylbaccatin III

O

OH

OOHO

HO

HO

OO

10

13

7

14

O

Paclitaxel R1= COC6H5 ; R2= CH3CODocetaxel R1= COOC(CH3)3 ; R2= H

O

OH

OOHO

OR2O

O

OO

O

OH

R1NH13

107

IDN 5109Licensed to Bayer and currently in Phase II

O

OH

OOO

OAcO

O

OO

O

OH

NH

O

O

OO

O

OH

OOHO

OHO

O

O

O

NH

OH

O

O O

Molecular Formula : C41H57NO14 M.W.: 787.38

NSC Number : 718373

[α]D = -51.0° (c= 1%, MeOH)

ANTITUMOR ACTIVITY OF IDN 5390ORALLY ADMINISTERED

IN MAMMARY CARCINOMA MX-1

Days after tumor implantation

Tum

or w

eigh

t (m

g)

0 10 20 30 40 50 60

1000

100

10

Control120 mg/kg qd5/w x 3 weeks 30 mg/kg 2qd5/w x 3 weeks

90 mg/kg 2qd5/w x 5 weeks 60 mg/kg 2qd5/w x 3 weeks

ANTINEOPLASTIC ACTIVITY OF IDN 5390ON 1A9/PTX22 HUMAN OVARIAN CARCINOMA

IDN 5390 120 mg/kg i.p. q1x5 for two weeks

0 10 15 20 25 30Days after tumor transplantation

Med

ian

tum

or w

eigh

t (g)

0.0

0.5

1.0

1.5

2.0

Treatment

Control

IDN 5390

Antitumor effect of oral IDN 5109 and IDN 5390,alone or in combination in Taconic SCID mice

implanted with human colon tumor xenograft, DLD-1

0 5 10 15 20 25 30 35 40 45 50 55 60 65

Med

ian

Tum

or V

olum

e (m

m3 )

0

200

400

600

800

1000

1200

1400

Days after DLD-1 tumor implant

Control

Vehicle Control IDN 5109 90 mg/kg (q7dx3)IDN 5390 60 mg/kg (qdx5/w x 4 weeks)

IDN 5109 270 mg/kg+ IDN 5390 1800 mg/kg

IDN 5390(NEW ANTICANCER DRUG)

Semisynthetic taxane derivative with modified baccatin skeleton•

Different mechanism of action•

Initially selected as inhibitor of angiogenesis it is endowed withan unknown mechanism

•

Active orally after chronical administration in several tumorxenograft and metastasis

•

Active on Paclitaxel-resistant cell lines•

Devoid of toxicity in repeated administration as body weight loss•

SELECTED ACTIVE PRINCIPLES AND HIGH CONTENT STANDARDIZED EXTRACTS FROM PLANTS

60% asiatic and madecassic acids,40% asiaticoside

70% flavonoids calculated as silybin

36% anthocyanosides

24% total ginkgoflavonglycosides,6% ginkgolides and bilobalide

14% ginsenosides as ginsenoside Rg1

25% hydroxyanthracene diglycosidescalculated as cascaroside A

95% polyphenols

45% sennosides calculated as sennoside B

13% total sterols calculated as ß-sitosterols

90% C8 to C20 linear chain fatty acids

0.3% hypericin

20% proanthocyanidins

3% luteolins glycosides

STANDARDIZATIONSelected triterpenes

Purified dry extract (silymarin)

Dry extract (Myrtocyan )

Dry extract

Dry extract

Selected hydroxyanthracene glycoside(Purselect ; Casanthranol)

Purified dry extract (LEUCOSELECTTM)

Purified dry extract

Purified soft extract

Purified extract

Methanolic extrac

Ethanolic extract

Ethanolic extract

PRODUCTCentella asiatica (L.) Urban

Silybum marianum (L.) Gaertn.

Vaccinium myrtillus L.

Ginkgo biloba L.

Panax ginseng C.A. Meyer

Rhamnus purshiana DC

Vitis vinifera L.

Senna alexandrina Miller

Prunus africana (Hook. f.) Kalkm

Serenoa repens (Bartr.) Small

Hypericum perforatum L.

Crataegus oxyacantha L.

Cynara scolymus L.

PLANTSerial parts

Fruits

Fruits

Leaves

Roots

Bark

Seeds

Leaves

Bark

Fruits

Leaves-flowers

Leaves-flowers

Leaves

PART

BIOLOGICALLY ACTIVE COMPOUNDSFROM PLANT KINGDOM

PURE PRINCIPLES MULTI COMPONENTS(raw drug, tinctures ...)

PHARMACEUTICAL MARKET

STANDARDIZED EXTRACTS,SELECTED FRACTIONS ...

PURE PRINCIPLES

DIETARYSUPPLEMENTS

F. Lang, H. Stumpf, PHARMEUROPA Vol. 11, No. 2, 268, June 1999 (adapted)

extract fillingheight

velocity offlow

staticalpressure

batchsize

method ofextraction

time ofextraction

extractiontemperature

extractionpressure

nature ofplant material

part ofplant material

origin ofplant material

degree ofprocessing

watercontent

nature ofsolvent

concentration ofsolvent

amount ofsolvent

STANDARDISATION

EMEA/HMPWG/9/99

Standardisation means adjusting the herbal drugpreparation to a defined content of a constituent or a groupof substances with known therapeutic activity respectivelyby adding excipients or by mixing herbal drugs or herbaldrug preparations (e.g. standardised extracts from theEuropean Pharmacopoeia).

*

* In some Member States the expression is used to describe all measures whichare taken during the manufacturing process and Quality Control leading to areproducible quality.

STANDARDISATIONPLANT MATERIAL QUALITY1.

MANUFACTURING PROCESS2.

IN PROCESS CONTROLS3.

GENERAL MANUFACTURING PROCESS SEQUENCE

QuarantineBIOMASSReleased / RejectedQUALITY CONTROL

GRINDINGEXTRACTIONPURIFICATION

QuarantineDRYINGReleased / RejectedQUALITY CONTROL

SHIPPING

In-process controls

GAPPRINCIPLES AND GUIDELINES FOR GOOD AGRICULTURAL PRACTICE

SEEDS AND PROPAGATION MATERIAL1.

Materials have to be identified botanically indicatingplant variety, cultivar, chemotype and origin.

CULTIVATION2.

Depending on the mode of cultivation e.g.conventional or organic, growers should be allowedto follow different SOP. In general, care should betaken to avoid environmental disturbances.The principles of good crop husbandry must befollowed including an appropriate rotation of crops.

HARVEST3.

Harvesting modalities and equipment have to bedescribed in details.

PRIMARY PROCESSING4.

Primary processing includes steps of processingsuch as washing, freezing, distilling, drying etc.All these processes whether for food or medicinaluse must conform to relevant European andNational Regulation.

PACKAGING5.

Modalities of packaging have to be described indetails.

STORAGE AND TRANSPORT6.

Modalities of storage, including dose for fresh andfrozen product have to be described in details.

EQUIPMENT7.

PERSONAL AND FACILITIES8.

Any starting material and processing steps have tobe documented, including the location of

cultivation.Each batch must have a Batch Packaging Record.Any mixing procedures should be documented byBatch Processing Records.

DOCUMENTATION9.

EDUCATION10.

Agreement between producers and buyers ofmedicinal and aromatic plants, with regard toquality issues, must be based on internationallyrecognized or national specifications and shouldbe documented in a written way.

QUALITY ASSURANCE11.

Self inspection should be conducted in order tomonitor the implementation and compliance withGAP. principles and to propose necessarycorrective measures.

SELF INSPECTION12.

Q7A

Q7A- GMP Concept• GMP controls apply to all steps in the

manufacturing process beginningwith the use of starting materials

• Only critical manufacturing steps aresubject to process validation

1. Introduction

2. Quality management

3. Personnel

4. Building and facilities

5. Process equipment

6. Documentation and records

7. Materials management

8. Production and in-process controls

9. Packaging and labelling

10. Storage and distribution

11. Laboratory controls

12. Validation

13. Change control

14. Rejection and reuse of materials

15. Complaints and recalls

16. Contract manufacturers and laboratories

17. API agents, brokers, distributors, repackers, and relabellers

18. Special biotech consideration

19. APIs for clinical trials

20. Glossary

13. Change control

Change control is a pivotal point.Every changement has to be evaluated inadvance also in relation to the industrial

impact.Normally in QA Departments a whole unit isdevoted to this particular problem.When the changement does not fit withacceptable criteria it is rejected.

GAP and GMP (ICH-Q7A)

allows the

correct standardisation of

pharmaceutical botanical derivatives

Echinacea angustifolia

E. paradoxa var. par.

E. angustifolia var. ang.

E. sanguinea

E. pallida

GENUS ECHINACEA

E. simulata

E. purpurea

E. atrorubens

CONFUSION

1. Leaves broadly to narrowly ovate, rounded at base, often serrate:

Echinacea purpurea

1. Leaves lanceolate to linear lanceolate. Attenuate to base, never serrate:

2. Ligules 4-9 cm long, white pollen: Echinacea pallida

2. Ligules 2-4 cm long, bright yellow pollen, compact growth habit,

max 0,5 m height: Echinacea angustifolia

CHEMICAL DIFFERENCES

Minutes5,00 10,00 15,00 20,00 25,00 30,00 35,00 40,00 45,00 50,00 55,00 60,00

Minutes10,00 20,00 30,00 40,00 50,00 60,00

EchinacosideAlkamide 8

E. angustifolia

E. pallidaEchinacoside

Cichoric acidAlkamide 8E. purpurea

Pentadeca-8Z-en-2-one

E. ANGUSTIFOLIATHE SELECTION: FIRST STEP

ANALYSIS

Polysaccharides %

Echinacoside %

Isobutylamides %

Resistance to parasites

IN VITROMICROPROPAGATION

SELECTED

PLANTS

E. ANGUSTIFOLIATHE SELECTION: SECOND STEP

CLONES

CULTIVATION AND PRODUCTION OF SEEDS

E. ANGUSTIFOLIATHE SELECTION: THIRD STEP

LARGE SCALE CULTIVATION

E. ANGUSTIFOLIA

BIOMASSFOREXTRACTION

FURTHER SELECTIONS AND MICROPROPAGATIONS

LARGE SCALE CULTIVATION

E. ANGUSTIFOLIA

Echinacea angustifolia rootsFirst hydroalcoholic extraction (for echinacoside)

Enriched extract(echinacoside)

Hydroalcoholic extract

Enriched extract(IDN 5405)

Second hydroalcoholicextraction

(for IDN 5405)

™

Wet plant material

Enriched fraction(isobutylamides)

n-hexanecounter-extraction

(for isobutylamides)

™

™

HO

CH CH COOHO OCH2

OCH2O

H

O

HO

HOOH

OHO O CH2 CH2

OH

OH

HO

O

HO

H3C

OH

O

OH

HO

OH

OH O

O

OH

OH

OH

CH2

O

O

OH

OH

OH

CH2

n

HO

LOW MOLECULAR WEIGHT POLYSACCHARIDE

Structure of inulin (fructose polymer)

hairy regionshairy regions

smooth regions

O

O

O

O

O

O CO2Me

HO

OH

HO

CO2H

OHCO2Ac

HO

OHO

smooth regions

The high molecular weight

polysaccharide is a methoxy pectin

with a backbone of

α-(1-4) polygalacturonic acid

partially carboxymethylated (about 60%)

and partially acetylated (about 9%)

(smooth region) and with hairy region of

substituted rhamnogalacturonan.

IDN 5405HIGH MOLECULAR WEIGHT POLYSACCHARIDE

POLYSACCHARIDES PROFILE

HIGH MOLECULAR

WEIGHT

LOW MOLECULAR

WEIGHT

IDN 5405

Inulin

ISOBUTYLAMIDES INECHINACEA ANGUSTIFOLIA ROOTS LIPOPHILIC EXTRACT

1 3 6

2 54

7 8

9

1210,11

1314

0 10 20 min

1) Undeca-2E,4Z-dien-8,10-diynoic acid isobutylamide2) Undeca-2E-en-8,10-diynoic acid isobutylamide3) Undeca-2Z,4E-dien-8,10-diynoic acid isobutylamide4) Undeca-2Z-en-8,10-diynoic acid isobutylamide5) Dodeca-2E-en-8,10-diynoic acid isobutylamide6) Trideca-2E,7Z-dien-10,12-diynoic acid isobutylamide7) Dodeca-2E,4Z,10Z-trien-8-ynoic acid isobutylamide

8) Undeca-2Z-en-8,10-diynoic acid 2-methylbutylamide9) Dodeca-2E-en-8,10-diynoic acid 2-methylbutylamide

10) Dodeca-2E,4E-8Z,10E-tetraenoic acid isobutylamide11) Dodeca-2E,4E-8Z,10Z-tetraenoic acid isobutylamide12) Pentadeca-2E,9Z-dien-12,14-diynoic acid isobutylamide13) Hexadeca-2E,9Z-dien-12,14-diynoic acid isobutylamide11) Dodeca-2E,4E-dienoic acid isobutylamide

OD

550

/650

0.5

0.4

0.3

0.2

0.1

0.0

0.6

0.8

0.7

Vehicle 11.1 33.3 100

Medium

Con A

µg/mL

CYTOTOXIC EFFECT ON MURINE SPLENOCYTES INDUCEDBY INCREASING CONCENTRATIONS OF ECHINACEA

ANGUSTIFOLIA ROOTS LIPOPHILIC EXTRACT

OD

550

/650

0.5

0.4

0.3

0.2

0.1

0.0

0.6

0.8

0.7

Vehicle 11.1 33.3 100

Medium

LPS

µg/mL

CYTOTOXIC EFFECT ON MURINE SPLENOCYTES INDUCEDBY INCREASING CONCENTRATIONS OF ECHINACEA

ANGUSTIFOLIA ROOTS LIPOPHILIC EXTRACT

EFFECT OF ECHINACEA ANGUSTIFOLIA ROOTS LIPOPHILIC EXTRACT

ON INTESTINAL MOTILITY IN MICE

Male Swiss mice (25-30 g) were fed on a charcoal meal (5% in 5% arabic gum).Animals were fasted for 3 hours before intraperitoneal administration of the compound.

Upp

er g

astr

oint

estin

al tr

ansi

t (%

) 60

50

40

30

20

10

0Control

p <0.01

**

E. ANGUSTIFOLIA ROOTS LIPOPHILIC EXTRACT5 mg/kg i.p.

ADDITIONAL INFORMATION ONECHINACEA ANGUSTIFOLIA ROOTS

LIPOPHILIC EXTRACT

Acute oral toxicity in mice: 236 mg/kg (LD50)

Electrophysiological experiments in rat sciaticnerve-EDL muscle: effects on nerve impulseand synaptic transmission

√

√

EFFECT OF Polinacea™ (ORAL ADMINISTRATION)ON IMMUNOCOMPETENT AND IMMUNOSUPPRESSED

CANDIDA ALBICANS INFECTED MICE

Treatment

Candida albicans (350,000 / i.v. / mouse)

Candida albicans + Polinacea™ (20 mg/mouse)

Candida albicans + Cyclosporin A (1 mg/mouse)

Candida albicans + Cyclosporin A + Polinacea™

Survivorsafter 10 days

20

20

10

10

0

6

0

4

n

Treatment Survivorsafter 13 days

Candida albicans 10 0

Candida albicans + Echinacin® 10 0

Candida albicans + LPS (40 ng/mouse) 10 1

Candida albicans + Polinacea™ (2 mg/mouse) 10 4

n

EFFECT OF Polinacea™ (I.P. ADMINISTRATION)ON IMMUNOCOMPETENT

CANDIDA ALBICANS INFECTED MICE

Control

100 µg/mL

10 µg/mLPolinacea™

LPS [ 1 µg/mL ]

5

3

3

3

Interferon-γ

56

45

79

15

Co-treatment ----- [ 1000 U/mL ]

1 µg/mL 3 21

0.01 µg/mL 3 11

(LPS : 200 ppm)

NO (µM) PRODUCTION BY J774 AFTER 48 HOURS OF TREATMENT

Control

Polinacea™

LPS [ 1 µg/mL ] 5.4

0

Interferon-γ

100 µg/mL 0 24.6

10 µg/mL 0 20.9

49.3

21.8

Co-treatment ----- [ 500 U/mL ]

1 µg/mL 0 21.4

0.1 µg/mL 0 16.1

0.01 µg/mL 0 25.9

NO (µM) PRODUCTION BY J774 AFTER 48 HOURS OF TREATMENT( ALL SAMPLES ARE LPS FREE)

Control

IDN 5405

LPS [ 1 µg/mL ] 6.4

0

Interferon-γ

100 µg/mL toxic toxic

10 µg/mL 0 18.0

50.0

16.8

Co-treatment ----- [ 500 U/mL ]

1 µg/mL 0 16.0

0.1 µg/mL 0 17.8

0.01 µg/mL 0 17.7

(high molecular weight polysaccharides)

NO (µM) PRODUCTION BY J774 AFTER 48 HOURS OF TREATMENT( ALL SAMPLES ARE LPS FREE)

Control

Inulin

LPS [ 1 µg/mL ] 14.6

0

Interferon-γ

100 µg/mL 0 10.0 (toxic)

10 µg/mL 0 13.9 (toxic)

51.8

24.6

Co-treatment ----- [ 500 U/mL ]

1 µg/mL 0 20.4

0.1 µg/mL 0 20.2

0.01 µg/mL 0 19.3

(low molecular weight polysaccharide)

NO (µM) PRODUCTION BY J774 AFTER 48 HOURS OF TREATMENT( ALL SAMPLES ARE LPS FREE)

PURIFIED SPLEEN T LYMPHOCYTES:IFN-γ PRODUCTION

3-wells mean values

Treatment 10 µg/mL 1 µg/mL 0.1 µg/mL

Medium ( 4.5 ± 0.5 )

αCD3 + IDN 5405

αCD3 + Polinacea™

αCD3 ( 149.5 ± 25.0 )

( ALL SAMPLES ARE LPS FREE)

690.5 ± 95.5

442.0 ± 70.5

466.3 ± 71.8

355.8 ± 61.4

410.0 ± 45.7

280.0 ± 35.8

MILESTONESSELECTED CULTIVATED ECHINACEA ANGUSTIFOLIA PLANTS

™

UNIQUE TRIPLE STANDARDIZATION

LOW LEVEL OF ISOBUTYLAMIDES

INNOVATIVE EXTRACTION PROCEDURE

DIRECT IMMUNOLOGICAL EFFECT ON T CELLS

WELL TOLERATED IN ACUTE AND SUBACUTE TOXICITY

IDEAL CANDIDATE FOR CLINICAL TESTING

Hypericum perforatum L.

0 20 40 60min

HPLC chromatogram of ST. JOHNSELECT™ (LI 160)

1

23

4 5

6

7

8 9

10

11

12

15

13

1-3 Chlorogenic acid analogues4-6,8,9 Flavonol derivatives7 Pentahydroxyflavanone10 I3,II8 Biapigenin11,12 Hypericin analogues13-16 Hyperforin analogues

14

16

ST. JOHNSELECT™ (LI 160)PERCENTAGE AMOUNT OF MAIN COMPONENTS

3.393.424.882.290.560.290.360.340.204.500.61

Percent (HPLC analysis)

Chlorogenic acidRutin

HyperosideIsoquercitrin

QuercitrinQuercetin

I3,II8 BiapigeninPseudohypericin

HypericinHyperforin

Adhyperforin

Component

From: BMJ 313, 253-258 (1996)

From: Linde K., Ramirèz G., Mulrow C.D., Pauls A., Weidenhammer W., Melchart D., BMJ 313, 253-258 (1996)

continue

From: Linde K., Ramirèz G., Mulrow C.D., Pauls A., Weidenhammer W., Melchart D., BMJ 313, 253-258 (1996)

VARIATION OF ANALYTICAL PARAMETERS OF ST. JOHN’S WORT PLANT MATERIAL AND PREPARATIONS

3.9 : 14.3 : 13.7 : 13.4 : 13.4 : 14.7 : 14.7 : 15.4 : 13.1 : 1

drug-extract-ratio

123456789

Lot

0.240.270.230.460.430.360.220.170.35

contentof

hypericin

0.581.551.843.523.857.778.309.59

13.68

contentof

hyperforin

preparations

content given as % values

0.0600.0620.0630.1360.1270.0760.0470.0320.122

0.1500.3610.4971.0351.1321.6531.7671.7765.025

contentof

hyperforin

plant materialcontent

ofhypericin

COMPARATIVE 13C-NMR OF TWOHYPERICUM PERFORATUM EXTRACTS

200 180 160 140 120 100 80 60 40 20 ppm

INDENA HYPERICUM PERFORATUM EXTRACTCOMMERCIAL HYPERICUM PERFORATUM EXTRACT

COMPARATIVE 1H-NMR OF TWOHYPERICUM PERFORATUM EXTRACTS

INDENA HYPERICUM PERFORATUM EXTRACTCOMMERCIAL HYPERICUM PERFORATUM EXTRACT

Characterization of Hypericum perforatum extracts byNIR (Near infrared spectroscopy)

H. PERFORATUM EXTRACT FROM DIFFERENT PRODUCERS

INDENA H. PERFORATUM STANDARDIZED EXTRACT

CONCLUSIONS

Standardisation of botanical derivatives according to GAP

and GMP (ICH Q7A) is the only procedure which

guarantees:

- the reproducibility of pharmacological and clinical effects;

- the complete assurance of reproducibility of safety.

Phytoequivalence can be evaluated by

combining classical analytical determination

(spectrophotometric, GC, HPLC) of known

products (markers, active principles) with an

innovative spectroscopic approach (13C-NMR,1H-NMR and NIR) which allows the qualitative

evaluation of the globality of the extracts.