Comunicação oral da qual só está disponível a apresentação.Possible international directive for quality control of bee pollen.Fundação Ciência e Tecnologia de Portugal
POCTI (FEDER) & COMPETE, Ciência Viv

Al-Lawatti, T.H.

Almaraz-Abarca, N.

Almeida-Muradian, L.

Anjos, O.

Barreto, L.

Boppré, M.

Campos, M.G.

Lopes, J.

Marcucci, M.C.

Nevers, V.

Poirot, B.

Stanciu, O.G.

Almeida-Muradian, L.B.

Barreto, Lídia M.R.C.

Repositório do Instituto Politécnico de Castelo Branco

Campos, M. G.1, Almaraz-Abarca N.2, Lopes J.,1 Anjos O.3, Marcucci M.C.4, Nevers V. 5, B. Poirot5, Al-Lawati T. H.6 , Almeida-MuradianL.7 Barreto L.8, Stanciu O. G.9, Boppré M.101 Center of Pharmaceutical Studies, Laboratory of Pharmacognosy, Faculty of Pharmacy, University of Coimbra, Polo III, Azinhaga de Santa Comba, Coimbra, 3000-548 Portugal - mgcampos@ff.uc.pt, joanalopes82@hotmail.com2Centro Interdisciplinario de Investigacio´n para el Desarrollo Integral Regional, Instituto Politécnico Nacional, Unidad Durango, Sigma s/n 20 de Noviembre II, Durango, México - noralab@yahoo.com3Polytechnic Institute of Castelo Branco/ Agrarian School, Quinta da Senhora de Mércules, Castelo Branco, 9001-909 Portugal -ofelia@ipcb.pt4Laboratório de Produtos Naturais, Programa de Mestrado em Farmácia da UNIBAN Rua Maria Candida, 1813 V. Guilherme, São Paulo, Brazil. cris.marcucci@yahoo.com.br5 APINOV,Pôle Technologique, 40, Rue Chef de Baie,17 000 La Rochelle France - valerie.nevers@apinov.com6Bee Science Research Laboratory at Plant Protection Research Center (PPRC), Ministére of Agriculture. Sultanate of Oman hassan98_2000@hotmail.com7Pharmaceutical Science School, University of São Paulo, Brazil ligiabic@gmail.com8Center of Research of Taubaté, University ,SP-Brasil - barretolidia@yahoo.com.br9Department of Beekeeping and Sericulture, University of Agricultural Sciences andVeterinary Medicine, 3-5 Manastur Street, 400151, Cluj-Napoca, Romania - ococan@yahoo.com10Forstzoologisches Institut, Albert-Ludwigs-Universität,D-79085 Freiburg i.Br.,Germany. boppre@fzi.uni-freiburg.dePOSSIBLE INTERNATIONAL DIRECTIVE FOR QUALITY CONTROL OF BEE POLLEN 42º INTERNATIONAL APICULTURAL CONGRESS APIMONDIA 2011 BUENOS AIRES  ARGENTINA   Congress: From 21st to 25th, September 2011Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra - Portugal                    Correspondence to: e-mail: mgcampos@ff.uc.ptMaria Graça CamposHead of the Bee Pollen Working Group at International Honey Comission (IHC)Specialist at European Scientific Cooperation Working Group on Isoflavones, at the European Community Agency, European Food Safety Authority  EFSA, (2009- ...) Specialist at Thecnical Comission of  Autoridade de Segurança Alimentar e Económica (ASAE/PORTUGAL) - Dietetic Products, Nutrition and allergies (2008-  )DRUG DISCOVERYQuality ControlRisk assessmentSUMARY Urgent need to have a International Directive for quality Control of Bee pollen to avoid International Trade Conflicts Bee pollen as Medicinal Plant (add value)Bioactivity correlacted to the floral source Data from various countries around the WorldDenomination by region or by floral source Future Challenges ~ ISO norm draft for BEE POLLENCampos & FrigerioHigh QualityCampos & FrigerioDRUGMedicinal PlantFUNCTIONAL FOODDIETARY SUPPLEMENTDepending of the floral originReunião Informal do Comité dos Medicamentos à Base de Plantas (HMPC) da Agência Europeia de Medicamentos (EMEA) - 27/28 Setembro 2007http://www.emea.europa.eu/Human Medicines - Herbal Medicinal ProductsDirective 2004/24/EC1 introduced a new pathway for marketing traditional herbal medicinal products, the "simplified registration".The Committee on Herbal Medicinal Products (HMPC) has been established in the framework of Directive 2004/24/EC and Regulation (EC) No 726/20042.The main task of the Committee is to establish Community herbal monographs and entries to the 'list of herbal substances, preparations and combinations thereof for use in traditional medicinal products'. Community monographs and entries to the Community List established by the HMPChttp://www.emea.europa.eu/Patients/herbals.htmCampos & FrigerioCampos & FrigerioMany research as been done in a way to prove the importance to explore the potential of bee-pollen for therapeutical purposesJournal of Health Science 52 (1) 43-49, 2006                ________________________________________________Anabolic effects of bee pollen Cistus ladaniferus extract on Bone components in the femoral dyaphiseal and metaphyseal tissues of rats in vitro and in vivo.Yamaguchi et al.,  JapanCampos & Frigerio Anaemia and the hemopoietic system  bee pollen - Iron Antioxidant activity  Eucalytus globulus  flavonoids + ?? Anti inflammatory activity - Pinus densiflora  flavonoids ?? Antitumoral activity  Brassica napus  polysaccharide LBPP Antiartherosclerotic activity - fat-soluble pollen extract Activity on osteoporosis  Cistus ladaniferus  PM < 1000 Activity in Disorders of the Prostate  Hidroxamic acid Antidiarrhoeal activity - Eucalytus globulus; Salix atrocinera Antimicrobial activity  Ranunculus sp  herbacetin (flavonoid)Floral origin  bioactivity compoundsTherapeutic window???Toxicity??Side effects???Toxic compounds can be used With therapeutic bioactivity Campos & FrigerioRISK ASSESSMENTData BaseAnalytical MethodsBee Pollen Standardization & QualityCampos & FrigerioIdentification of the worldsmain floral sourcesMicroscopic analysis (Barth et al, 2010 Anales de Academia Brasileira de Ciências, 82(4)HPLC/DAD phenolic profiles(Campos et al, 1996 Phytochemical Analysis)050100% Mobile Phase00.20.4AU0 20 40 60Minutesc:\amine\hplc\magui\estroge3.001\estroge3.gdt : 340nm, 0.01 sensitivity : amostra inteira: Inj. Number: 2c:\amine\hplc\magui\estroge3.001\estroge3.gdt : 340nm, 0.01 sensitivity : carga polinica: Inj. Number: 3Bee pollen pellet from Eucalyptus(100% taxon pure)Sample with Eucalyptus as major source100%60%% 0f the major floral origin in a mixtureFloral origin determinationCampos & FrigerioPORTUGALBRASILBULGARIAPhenolic and polyphenolic compunds= fingerprintM.G. CamposCampos M. G., Webby R. F., Markham K. R.,  2002. The Unique Occurrence of the Flavone Aglycone Tricetin in Myrtaceae Pollen.  Z. Naturforsch. 57c, 944-946  Markham K., Mitchel K., Campos M. G., 1997. An unusually lipophilic flavonol glycoside from Ranunculus sardous Crantz. pollen -Phytochemistry, 45 (1), 203-204.Markham K., Campos M.G., 1996. 7- and 8-O-methylherbacetin-3-O-sophoroside from bee pollens and some struture/activity observations. Phytochemistry, 43 (4) 762-767.Polyphenolic profilesIdentification of the taxonAnalytical methodsUlex europeaus Bee pollen loadsPortugal 1992 Portugal 1992 Pollen from plantNew Zealand 1995New Zealand 1994Campos & FrigerioFingerprint analysisUV-VisNew ZealandCampos & FrigerioFingerprint analysisOOOHOOHOHOH- glu - glu(6-1)rhaOOOHOOHOHKaempferol-3-O-sophorosideOglu -OOOOHOH - gluKaempferol-3,7-O-diglucoside    OOOH3COOHOHOH - glu(2-1)glu7-O-methylherbacetin-3-O-sophorosideM.G. CamposBAND II BANDIb    aCampos M. G., Markham K. R. Structure information from HPLC and on-line measure absorption spectra  Flavones, Flavonols and Phenolic Acids Ed. Imprensa da Universidade de Coimbra 2007BAND II BAND Iquercetin-3-O-glucosideHPLC/DAD/MS/NMR  PortugalNew Zealand Rare compoundsRomaniaLife Sciences InstituteUniversity of Agricultural Sciences and Veterinary MedicineCluj-NapocaRomaniaPolyphenolic fingerprint analysisSalix sp. bee pollenKaempferol-3-O-neohesperidosideCaffeic acid derivativeSlovakiaEva MolnarovaCampos & FrigerioCampos & FrigerioMEXICOBee pollen production in BrasilCENTER OF RESEARCH OF TAUBATÉ UNIVERSITY-SP-BRASILLEVANTAMENTO DE BARRETO &ORSI 2010 =19 ESTADOS E DISTRITO FEDERAL=20, BA+SE+SCGood Practice Bee Pollen Quality Standard BookletMULTIFLORAL(native flora)Production area (a),  Bee Pollen details (b),    Drying heater (c),        Packaging (d) Bee Pollen Grains (Optic Microscopy 400x) Mean ± DPMoisture % 5.05±1.54Ash% 3.35±0.77Proteins % 14.64±3.67Total sugars % 31.82±2.25Fiber % 4.21±0.35Free acidity mEq 246.50±41.24pH 4.46±0.28Lipids % 3.82±2.10Water activity 0.32±0.01Vitamin E  mcg/g 6.05±0.37Total Phenolics % 0.23±0.02Total Flavonoids mg/g 4.64±0.07Future ChallengesUniversity of Coimbra/Faculty of PharmacyHealth Sciences Campus - PortugalThe authors wish to thank Fundação Ciência e Tecnologia de Portugal Our projects are funded by POCTI (FEDER) & COMPETE, Ciência Vivamgcampos@ff.uc.ptThanks to All my Coleaguesthat work with Bee Pollen

Possible international directive for quality control of bee pollen

Repositório Científico do Instituto Politécnico de Castelo Branco

Campos, M. G.1, Almaraz-Abarca N.2, Lopes J.,1 Anjos O.3, Marcucci M.C.4, Nevers V. 5, B. Poirot5, Al-Lawati T. H.6 , Almeida-Muradian
L.7 Barreto L.8, Stanciu O. G.9, Boppré M.10
1 Center of Pharmaceutical Studies, Laboratory of Pharmacognosy, Faculty of Pharmacy, University of Coimbra, Polo III, Azinhaga de 
Santa Comba, Coimbra, 3000-548 Portugal - mgcampos@ff.uc.pt, joanalopes82@hotmail.com
2Centro Interdisciplinario de Investigacio´n para el Desarrollo Integral Regional, Instituto Politécnico Nacional, Unidad Durango, Sigma s/n 
20 de Noviembre II, Durango, México - noralab@yahoo.com
3Polytechnic Institute of Castelo Branco/ Agrarian School, Quinta da Senhora de Mércules, Castelo Branco, 9001-909 Portugal -
ofelia@ipcb.pt
4Laboratório de Produtos Naturais, Programa de Mestrado em Farmácia da UNIBAN Rua Maria Candida, 1813 V. Guilherme, São Paulo, 
Brazil. cris.marcucci@yahoo.com.br
5 APINOV,Pôle Technologique, 40, Rue Chef de Baie,17 000 La Rochelle France - valerie.nevers@apinov.com
6Bee Science Research Laboratory at Plant Protection Research Center (PPRC), Ministére of Agriculture. Sultanate of Oman 
hassan98_2000@hotmail.com
7Pharmaceutical Science School, University of São Paulo, Brazil ligiabic@gmail.com
8Center of Research of Taubaté, University ,SP-Brasil - barretolidia@yahoo.com.br
9Department of Beekeeping and Sericulture, University of Agricultural Sciences and
Veterinary Medicine, 3-5 Manastur Street, 400151, Cluj-Napoca, Romania - ococan@yahoo.com
10Forstzoologisches Institut, Albert-Ludwigs-Universität,D-79085 Freiburg i.Br.,Germany. boppre@fzi.uni-freiburg.de
POSSIBLE INTERNATIONAL DIRECTIVE FOR QUALITY 
CONTROL OF BEE POLLEN 
42º INTERNATIONAL APICULTURAL CONGRESS APIMONDIA 2011 
BUENOS AIRES  ARGENTINA   Congress: From 21st to 25th, September 2011
Center of Pharmaceutical Studies, 
Faculty of Pharmacy, University of Coimbra - Portugal                    
Correspondence to: e-mail: mgcampos@ff.uc.pt
Maria Graça Campos
Head of the Bee Pollen Working Group at International Honey Comission (IHC)
Specialist at European Scientific 
Cooperation Working Group on Isoflavones, at the 
European Community Agency, European Food 
Safety Authority  EFSA, (2009- ...) 
Specialist at Thecnical Comission of  Autoridade 
de Segurança Alimentar e Económica 
(ASAE/PORTUGAL) - Dietetic Products, 
Nutrition and allergies (2008-  )
DRUG DISCOVERY
Quality Control
Risk assessment
SUMARY
 Urgent need to have a International Directive for 
quality Control of Bee pollen to avoid International 
Trade Conflicts
 Bee pollen as Medicinal Plant (add value)
Bioactivity correlacted to the floral source
 Data from various countries around the World
Denomination by region or by floral source
 Future Challenges ~ ISO norm draft for BEE POLLEN
Campos & Frigerio
High Quality
Campos & Frigerio
DRUG
Medicinal Plant
FUNCTIONAL FOOD
DIETARY SUPPLEMENT
Depending of the floral origin
Reunião Informal do Comité dos Medicamentos à Base de Plantas (HMPC) da Agência 
Europeia de Medicamentos (EMEA) - 27/28 Setembro 2007
http://www.emea.europa.eu/
Human Medicines - Herbal Medicinal Products
Directive 2004/24/EC1 introduced a new pathway for 
marketing traditional herbal medicinal products, the 
"simplified registration".
The Committee on Herbal Medicinal Products (HMPC) has been established in the 
framework of Directive 2004/24/EC and Regulation (EC) No 726/20042.
The main task of the Committee is to establish Community herbal monographs and 
entries to the 'list of herbal substances, preparations and combinations thereof for 
use in traditional medicinal products'. Community monographs and entries to the 
Community List established by the HMPC
http://www.emea.europa.eu/Patients/herbals.htm
Campos & Frigerio

Campos & Frigerio
Many research as been done in a way to prove 
the importance to explore the potential of 
bee-pollen for therapeutical purposes
Journal of Health Science 52 (1) 43-49, 2006                
________________________________________________
Anabolic effects of bee pollen Cistus ladaniferus extract on Bone 
components in the femoral dyaphiseal and metaphyseal tissues of 
rats in vitro and in vivo.
Yamaguchi et al.,  Japan
Campos & Frigerio
 Anaemia and the hemopoietic system  bee pollen - Iron
 Antioxidant activity  Eucalytus globulus  flavonoids + ??
 Anti inflammatory activity - Pinus densiflora  flavonoids ??
 Antitumoral activity  Brassica napus  polysaccharide LBPP
 Antiartherosclerotic activity - fat-soluble pollen extract
 Activity on osteoporosis  Cistus ladaniferus  PM < 1000
 Activity in Disorders of the Prostate  Hidroxamic acid
 Antidiarrhoeal activity - Eucalytus globulus; Salix atrocinera
 Antimicrobial activity  Ranunculus sp  herbacetin (flavonoid)
Floral origin  bioactivity compounds
Therapeutic window???
Toxicity??
Side effects???

Toxic compounds can be used 
With therapeutic bioactivity 
Campos & Frigerio
RISK ASSESSMENT
Data Base
Analytical Methods
Bee Pollen Standardization & Quality
Campos & Frigerio
Identification of the worlds
main floral sources
Microscopic analysis 
(Barth et al, 2010 Anales de Academia Brasileira de Ciências, 82(4)
HPLC/DAD phenolic profiles
(Campos et al, 1996 Phytochemical Analysis)

050
100
%
 M
ob
ile
 P
ha
se
0
0.2
0.4
AU
0 20 40 60
Minutes
c:\amine\hplc\magui\estroge3.001\estroge3.gdt : 340nm, 0.01 sensitivity : amostra inteira: Inj. Number: 2
c:\amine\hplc\magui\estroge3.001\estroge3.gdt : 340nm, 0.01 sensitivity : carga polinica: Inj. Number: 3
Bee pollen pellet from Eucalyptus
(100% taxon pure)
Sample with Eucalyptus as major source
100%
60%
% 0f the major floral origin in a mixture
Floral origin determination
Campos & Frigerio
PORTUGAL
BRASIL
BULGARIA
Phenolic and polyphenolic compunds
= fingerprint
M.G. Campos
Campos M. G., Webby R. F., Markham K. R.,  2002. The Unique Occurrence of the Flavone Aglycone Tricetin in Myrtaceae Pollen.  
Z. Naturforsch. 57c, 944-946  
Markham K., Mitchel K., Campos M. G., 1997. An unusually lipophilic flavonol glycoside from Ranunculus sardous Crantz. pollen -
Phytochemistry, 45 (1), 203-204.
Markham K., Campos M.G., 1996. 7- and 8-O-methylherbacetin-3-O-sophoroside from bee pollens and some struture/activity 
observations. Phytochemistry, 43 (4) 762-767.
Polyphenolic profiles
Identification of the taxon
Analytical methods
Ulex europeaus 
Bee pollen loads
Portugal 
1992 
Portugal 
1992 
Pollen from plant
New Zealand 
1995
New Zealand 
1994
Campos & Frigerio

Fingerprint analysis
UV-Vis
New Zealand
Campos & Frigerio
Fingerprint analysis

OO
O
HO
OH
OH
OH
- glu
 - glu(6-1)rha
O
O
O
HO
OH
OH
Kaempferol-3-O-sophoroside
Oglu -
O
O
O
OH
OH
 - glu
Kaempferol-3,7-O-diglucoside    
O
O
O
H3CO
OH
OH
OH
 - glu(2-1)glu
7-O-methylherbacetin-3-O-sophoroside
M.G. Campos
BAND II BANDI
b    a
Campos M. G., Markham K. R. Structure information from HPLC and on-line measure absorption spectra  Flavones, 
Flavonols and Phenolic Acids Ed. Imprensa da Universidade de Coimbra 2007
BAND II BAND I
quercetin-3-O-glucoside
HPLC/DAD/MS/NMR  
Portugal
New Zealand 
Rare compounds
Romania
Life Sciences Institute
University of Agricultural Sciences 
and Veterinary Medicine
Cluj-Napoca
Romania
Polyphenolic fingerprint analysis
Salix sp. bee pollen
Kaempferol-3-O-neohesperidoside
Caffeic acid derivative
SlovakiaEva Molnarova
Campos & Frigerio
Campos & Frigerio
MEXICO
Bee pollen production in Brasil
CENTER OF RESEARCH 
OF TAUBATÉ 
UNIVERSITY-SP-BRASIL
LEVANTAMENTO DE BARRETO &ORSI 2010 =
19 ESTADOS E DISTRITO FEDERAL=20, BA+SE+SC
Good Practice 
Bee Pollen Quality Standard Booklet
MULTIFLORAL
(native flora)

Production area (a),  Bee Pollen details (b),    Drying heater (c),        Packaging (d) 
Bee Pollen Grains (Optic Microscopy 400x) 



Mean ± DP
Moisture % 5.05±1.54
Ash% 3.35±0.77
Proteins % 14.64±3.67
Total sugars % 31.82±2.25
Fiber % 4.21±0.35
Free acidity mEq 246.50±41.24
pH 4.46±0.28
Lipids % 3.82±2.10
Water activity 0.32±0.01
Vitamin E  mcg/g 6.05±0.37
Total Phenolics % 0.23±0.02
Total Flavonoids mg/g 4.64±0.07
Future Challenges
University of Coimbra/Faculty of Pharmacy
Health Sciences Campus - Portugal
The authors wish to thank Fundação Ciência e Tecnologia de Portugal 
Our projects are funded by POCTI (FEDER) & COMPETE, Ciência Viva
mgcampos@ff.uc.pt
Thanks to All my Coleagues
that work with Bee Pollen


English

https://repositorio.ipcb.pt/bitstream/10400.11/1960/1/possible_international_directive_for_quality_control_of_bee_pollen-mara_g_campos.pdf

Possible international directive for quality control of bee pollen

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Repositório do Instituto Politécnico de Castelo Branco

Repositório Científico do Instituto Politécnico de Castelo Branco