Dr.
Hartwig
& Co. KG
Name
derSievers,
Firma /PhytoLab
Titel derGmbH
Präsentation
ANVISA Workshop
Equivalência de Extratos Vegetais
Requisitos regulatórios de documentação em caso
de mudança do fabricante do extrato vegetal
29 September 2015, Brasilia
Hartwig Sievers, PhytoLab
Topics
1. Regulatory requirements
a.
b.
c.
d.
2.
RESOLUTION - RDC No. 26, DATED MAY 13, 2014
INSTRUÇÃO NORMATIVA Nº 4, 18 JUNHO 2014
European Commission (EC) No 1234/2008
European Commission (2013/C 223/01)
Examples and documentation
a.
b.
Sourcing and production
Quality of the herbal drug/drug preparation
Residues and Contaminants
Markers
Constituents with relevance for safety
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
2
Variations – EU Regulation
COMMISSION REGULATION (EC) No 1234/2008
concerning the examination of variations to the terms of marketing
authorisations for medicinal products
(3) Variations to medicinal products can be classified in different
categories, depending on the level of risk to public or animal health
and the impact on the quality, safety and efficacy of the medicinal
product concerned.
Definitions for each of those categories should therefore be laid
down.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Variations – EU Regulation
COMMISSION REGULATION (EC) No 1234/2008
concerning the examination of variations to the terms of marketing
authorisations for medicinal products
European Commission (2013/C 223/01)
Guidelines on the details of the various categories of variations, …
concerning the examination of variations to the terms of marketing
authorisations for medicinal products … and on the documentation to
be submitted pursuant to those procedures
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Variations: Classification Guideline (2013/C 223/01)
B. QUALITY CHANGES
B.I ACTIVE SUBSTANCE
B.I.a) Manufacture
B.I.a.1 Change in the manufacturer of a starting
Conditions to
material/reagent/intermediate used in the
be fulfilled
manufacturing process of the active substance or
change in the manufacturer (including where relevant
quality control testing sites) of the active substance,
where no Ph. Eur. Certificate of Suitability is part of
the approved dossier.
Documentation
to be supplied
g. Introduction of a new manufacturer of the active
substance that is not supported by an ASMF and
requires significant update to the relevant active
substance section of the dossier
Procedure
type
II
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Variations: Classification Guideline (2013/C 223/01)
Documentation
1. Amendment of the relevant section(s) of the dossier (presented in the EU-CTD
format or NTA volume 6B format for veterinary products, as appropriate), if
applicable.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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CTD Module 3.2.S Drug Substance
3.2.S.1
General Information
3.2.S.2
Manufacture
3.2.S.2.1
Manufacturer(s)
3.2.S.2.2
Description of manufacturing process and process controls
3.2.S.2.3
Control of materials
3.2.S.2.4
Controls of critical steps and intermediates
3.2.S.2.5
Process validation and/or evaluation
3.2.S.2.6
Manufacturing process development
3.2.S.3
Characterisation
3.2.S.4
Control of drug substance
3.2.S.4.1
Specification
3.2.S.4.2
Analytical Procedures
3.2.S.4.3
Validation of analytical procedures
3.2.S.4.4
Batch analyses
3.2.S.4.5
Justification of Specification
3.2.S.5
Reference Standards or Materials
3.2.S.6
Container Closure System
3.2.S.7
Stability
3.2.S.7.1
Stability summary and conclusions
3.2.S.7.2
Post-approval stability protocol and stability commitment
3.2.S.7.3
Stability data
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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INSTRUÇÃO NORMATIVA Nº 4, 18 JUNHO 2014
2.1 DETALHES DA COLETA/COLHEITA E CONDIÇÕES DE CULTIVO
detalhes da coleta/colheita no laudo de análise da droga vegetal, como
por exemplo:
• data da coleta (XX/XX/XXXX)
• período do dia coletado (manhã, tarde ou noite)
• local de coleta (Cidade-Estado e coordenadas de GPS)
• condições do tempo no momento da coleta (nublado, ensolarado,
garoa)
• fase de desenvolvimento da planta (vegetativo, floração, frutificação,
maturação)
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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INSTRUÇÃO NORMATIVA Nº 4, 18 JUNHO 2014
2.2 ESTABILIZAÇÃO, SECAGEM E CONSERVAÇÃO
Na etapa seguinte, é importante descrever o modo de secagem: se foi
um processo natural (à sombra, ao sol ou mista - sol e sombra) ou
artificial (p. ex. circulação de ar, aquecimento, aquecimento com
circulação de ar, vácuo, esfriamento), a temperatura, o tempo de
secagem e o volume que foi seco. A última etapa, a conservação,
engloba a estocagem, embalagem e a manutenção das drogas após a
embalagem, sendo necessário informar condições de luminosidade,
umidade e temperatura.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Drying conditions and caffeoyl quinic
acids content in Artichoke leaves
120
100
97
100
92
relative content %
85
80
60
60
40
40
20
10
0
25
30
35
acc. to Hannig, Eich, 2001
40
45
50
55
60
Drying temperature ( °C )
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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RESOLUTION - RDC No. 26/14, Subsection I
Vegetable drug
Art. 13. An analysis report of the vegetable drug shall be presented, indicating
the method used, the specification and the results obtained for a batch of the
tests described below:
V - purity and integrity tests, including:
…
e) determination of heavy metals;
f) determination of residues from agrotoxic agents and the like;
i) determination of mycotoxin, to be performed when the need of such
evaluation or reports of contamination of species by mycotoxins are
mentioned in technical-scientific documentation
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Ph.Eur.: Herbal Drugs - TESTS
Pesticides (2.8.13). Herbal drugs comply with the requirements for pesticide residues.
The requirements take into account the nature of the plant, where necessary the
preparation in which the plant might be used, and where available the knowledge of the
complete record of treatment of the batch of the plant.
Heavy metals (2.4.27). Unless otherwise stated in an individual monograph or unless
otherwise justified and authorised:
– cadmium: maximum 1.0 ppm;
– lead: maximum 5.0 ppm;
– mercury: maximum 0.1 ppm.
Where necessary, limits for other heavy metals may be required.
Where necessary herbal drugs comply with other tests, such as the following, for
example.
Aflatoxin B1 (2.8.18). Where necessary, limits for aflatoxins may be required.
Ochratoxin A (2.8.22). Where necessary, a limit for ochratoxin A may be required.
Radioactive contamination. In some specific circumstances, the risk of radioactive
contamination is to be considered.
Microbial contamination. Where a herbal drug is used whole, cut or powdered as an
ingredient in a medicinal product, the microbial contamination is controlled (5.1.8.
Microbiological quality of herbal medicinal products for oral use and extracts used in their
preparation or 5.1.4. Microbiological quality of non-sterile pharmaceutical preparations
and substances for pharmaceutical use.
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29 Sep 2015
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Ph.Eur.: Herbal drug extracts - TESTS
Where applicable, as a result of analysis of the herbal drug used for
production and in view of the production process, tests for
microbiological quality (5.1.4 or 5.1.8), heavy metals (2.4.27),
aflatoxins (2.8.18), ochratoxin A (2.8.22) and pesticide residues
(2.8.13) in the extracts may be necessary. Where a test for heavy metals is
carried out, the same limits for heavy metals as those given in the
monograph Herbal drugs (1433) are applicable to extracts unless
otherwise stated in an individual extract monograph or unless otherwise
justified and authorized.
Herbal drug extracts - PRODUCTION
Where justified, herbal drugs used for the production of extracts may exceed the limits
for heavy metals specified in the monograph Herbal drugs (1433) provided that the
resulting extract satisfies the requirements for heavy metals (see Tests).
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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INSTRUÇÃO NORMATIVA Nº 4, 18 JUNHO 2014
2.4.8 Micotoxinas
... . As micotoxinas são normalmente compostos oriundos do
metabolismo secundário de fungos, sendo os mais comumente
relatados os dos gêneros Aspergillus, Fusarium e Penicillium, (OMS,
2007), compreendendo quatro principais grupos: aflatoxinas,
ocratoxinas, fumonisinas e tricotecenos, todos com efeitos tóxicos
(Silveira et al., 2010; Santos et al., 2013)
H. Sievers, ANVISA Workshop Brasilia
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Contaminants
Examples for relevance of heavy metals, mycotoxins
Hypericum perforatum
Salix spec.
Viscum album
Glycyrrhiza spec.
Cassia spec.
Cd
Cd
Cd
Ochratoxin A
Aflatoxin A
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Monitoring results (Ochratoxin A)
based on data from ca. 2000 samples of herbal drugs
occasional and frequent positive results (> 20 µg/kg):
Angelica root, Nettle root, Ginkgo leaf, Ononis spinosa root, Ginger root,
Cocoa, Taraxacum root, Orange flowers, Pepper, Licorice root
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Effect of GACP on Ochratoxin A content
in Licorice root (µg/kg)
year
n
Max. value
90th Percentile
1999
50
425
234
2000
159
904
145
2001
192
337
41
2002
152
423
34
2003
60
72
20
2010
257
117
26
2011
82
65
19
Klier
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Ochratoxin A, maximum levels
product
maximum µg/kg
source
Cereals (commodity)
Raisins
Instant Coffee
Roasted Coffee Beans
5,0
10,0
10,0
5,0
(EC) No 1881/2006
Wine, Grape Juice
2,0
(EC) No 1881/2006
15 (since 1.7.2012)
(EC) No 1881/2006
0,50
20
80
(EC) No 1881/2006
Various spices
Infant food
Licorice root (herbal tea)
Licorice extract (4:1)
Klier
(EC) No 1881/2006
(EC) No 1881/2006
(EC) No 1881/2006
(EC) No 1881/2006 bzw. Ph.Eur.
(EC) No 1881/2006 bzw. Ph.Eur.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Ochratoxin A in Licorice root, HPLC-UV
Ochratoxin A
Column: RP 18, Water - Acetonitrile, Detector: Fluorescence (Ex 330 nm, Em 460 nm) sample prep
SPE (immunoaffinity) , LFGB Coffee
0
10
Time (min)
20
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Mass spectrum Ochratoxin A
Ochratoxin A - MS/MS, Optimierung 030400
030400ochratoxin007 21 (0.368) Cm (2:48)
Daughters of 402ES1.54e6
358
100
COOH
O
N
H
OH
O
O
%
Cl
402
211
253
166
0
50
100
150
200
250
313 322
300
350
400
450
m/z
500
H. Sievers, ANVISA Workshop Brasilia
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Ochratoxin A in Licorice extract selective analysis by HPLC / MS / MS
Column: RP 18, Eluent: Wasser Acetonitrile, Detection: ESI(-), MRM m/z 402 - m/z 358
2 4 0 2 o c h ra to x in 0 1 6 S m (S G , 2 x 2 )
relative Intensity (%)
M R M of 1 C hannel ES402 > 358
4 .4 4 e 4
7 .9 2
100
Ochratoxin A
178 ppb
%
0
1 .0 0
2 .0 0
3 .0 0
4 .0 0
5 .0 0
6 .0 0
7 .0 0
8 .0 0
9 .0 0
1 0 .0 0
1 1 .0 0
1 2 .0 0
1 3 .0 0
T im e
1 4 .0 0
Time (min)
H. Sievers, ANVISA Workshop Brasilia
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Ochratoxin A, risk management
targeted analyses (risk assessment based)
Ochratoxin A is soluble in alcoholic solvents – for herbal drug at risk of
contamination the transition into the extract should be investigated
only few herbs/drugs are at risk of contamination, for other herbs
routine testing is not required
if no limits from Pharmacopoeia or Food regulation are
applicable/feasible, limits have to be established considering
toxicology, dosage and the ALARA principle (As Low As Reasonably
Achievable)
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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RESOLUTION - RDC No. 26, DATED MAY 13, 2014
Section II, Scope
Art. 2
This Resolution applies to industrialized products, which fit the categories
of phytotherapeutic drugs and traditional phytotherapeutic products.
§3
The traditional phytotherapeutic products cannot refer to diseases,
disorders, conditions or actions considered severe, cannot contain raw
materials at concentration with known toxic risk
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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RESOLUTION - RDC No. 26, DATED MAY 13, 2014
Subsection II Traditional phytotherapeutic products
Art. 23.
The traditional use shall be proven by means of technical-scientific
documentations, which shall be evaluated according to the following
criteria:
…
V - lack of AVPI with known toxic risk or toxic chemical groups or
substances at concentration higher that the proven safe limits;
H. Sievers, ANVISA Workshop Brasilia
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INSTRUÇÃO NORMATIVA Nº 4
2.3.2 Identificação química
B) Interpretação do perfil cromatográfico
A extensão permitida de variação no perfil cromatográfico deverá ser
determinada caso a caso. Isso acontece porque pequenas variações
podem ser importantes, especialmente se a variação estiver associada
com a presença de uma ou mais substâncias tóxicas. Há, porém, casos
em que uma variação maior pode não ser significativa.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Compounds with safety relevance
Examples for AVPI with compounds relevant for safety:
Artemisia absinthium
Salvia officinalis
Petasites hybridus
Symphytum officinale
Angelica officinalis
Illicium verum
Hypericum perforatum
Acorus calamus
Thujone
Thujone
Pyrrolizidine alcaloids
Pyrrolizidine alcaloids
Furanocumarins
Anisatin (adulteration)
Hyperforin (interactions)
Asarone
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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RDC No. 26/14, Art. 13. X – Marker analysis
Subsection I Vegetable drug
Art. 13. An analysis report of the vegetable drug shall be presented, indicating the
method used, the specification and the results obtained for a batch of the tests
described below:
VI - details about the harvesting/crop and the cultivation conditions, when
cultivated;
…
IX - chromatographic profile, accompanied by the respective image in an
electronic file recognized by Anvisa, with comparison that can guarantee the
identity of the vegetable drug; and
X - quantitative analysis of the marker(s) or biological control.
H. Sievers, ANVISA Workshop Brasilia
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RDC No. 26/14, Art. 13. X – Marker analysis
Example:
Humulus lupulus
conservative marker: xanthohumol
but: aqueous hop extract does not contain xanthohumol
alternative marker: hulupinic acid
but: constant trait?
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Art. 13. X – Marker analysis
0 2 0 3 0 0 h o p fe n -6 S m (M n , 2 x 1 ); S b (1 ,3 0 .0 0 )
3 .0 1
100
1: S can ES 353+339
1 .6 1 e 5
2 4 .4 1
4 .4 3
Xanthohumol
relative intensity [%]
Isoxanthohumol
2 2 .7 5
1 4 .4 7
%
3 .4 0
0 .9 6
4 .7 8
1 .9 2
1 4 .0 9
8 .0 0
1 0 .2 3 1 1 .0 1
5 .3 5 7 .5 8
5 .7 0
1 5 .8 5
1 7 .0 2
1 9 .1 4
2 6 .0 8
1 1 .8 6
2 1 .3 0
2 8 .4 8 2 9 .7 9
2 7 .6 3
3 4 .4 6
3 1 .7 0 3 4 .1 1
3 5 .1 3
3 8 .0 7
0
0 2 0 3 0 0 h o p fe n -6
3 9 .7 6
1: S can ES T IC
1 .8 8 e 7
1 7 .3 8
100
3 8 .6 7
Marker = hulupinic acid
2 .8 7
3 .0 5
%
4 .3 9
2 .2 0
5 .4 2
2 9 .6 8
6 .3 7
7 .6 1
0 .8 2
2 1 .8 7
1 2 .7 8
9 .7 3 1 0 .5 8
1 1 .9 6
2 6 .1 1
3 0 .9 6
0
2 .5 0
5 .0 0
7 .5 0
time [min]
1 0 .0 0
1 2 .5 0
1 5 .0 0
1 7 .5 0
2 0 .0 0
2 2 .5 0
2 5 .0 0
2 7 .5 0
3 0 .0 0
3 2 .5 0
3 5 .0 0
3 7 .5 0
T im e
4 0 .0 0
H. Sievers, ANVISA Workshop Brasilia
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Art. 13. X – Marker analysis
trace1:
hulupinic acid
trace 2 bis 6:
hop raw materials
hulupinic acid
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29 Sep 2015
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Art. 13. X – Marker analysis
Specification of different Hypericum products
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Art. 13. X – Marker analysis
Hyperforin (%) content in the extract
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Processing technology/galenic preparation
Water or ethanol – is it the same?
no interaction potential
Water
Hyperforin
interaction potential
Ethanol
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Passiflora incarnata
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29 Sep 2015
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Flavonol content in Passiflora spec.
Chromatographic conditions
HPLC-system
UHPLC H-class (Waters)
Column
ACQUITY UPLC® BEH C18 100 x 2.1mm, 1.7 µm
Column
temperature [°C]:
44
Injector temperature
[°C]:
Detection
wavelength:
25
220 – 500 nm,
Evaluation at 338 nm (apigenin derivatives)
Evaluation at 348 nm (luteolin derivatives)
Injection volume
[µl]:
Eluent A (v/v):
1.0
Eluent D (v/v):
Methanol : acetonitrile : formic acid (750 : 250.: 1,
v/v/v)
water : formic acid (1000 : 1, v/v)
Gradient:
Gradient Table
T ime
(min)
Fl ow Rate
(mL/min)
%A
%B
%C
%D
1
Initial
0.270
90.0
0.0
0.0
10.0
2
15.00
0.270
77.5
0.0
0.0
22.5
3
20.00
0.270
10.0
0.0
0.0
90.0
4
25.00
0.270
10.0
0.0
0.0
90.0
5
25.50
0.270
90.0
0.0
0.0
10.0
6
30.00
0.270
90.0
0.0
0.0
10.0
H. Sievers, ANVISA Workshop Brasilia
H. Sievers, ANVISA Workshop Brasilia
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Fingerprint P. incarnata (USA)
USA: isovitexin-glucoside is the dominant compound
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Fingerprint P. incarnata (Eur 1)
Europe: isovitexin is the most dominant compound
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
37
Fingerprint P. incarnata (Eur 2)
Europe: isovitexin is the most dominant compound
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Discriminant analysis
Source
Italy
France
North America
European
type2
Austria
Spain
Hungary
European
type1
North
American
type
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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TLC Fingerprint Passiflora incarnata
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Markers – Influence of Production process
Herbal raw material
drying conditions (e.g., Cynara)
Herbal preparation
extraction conditions, e.g., temperature (e.g., Cynara)
excipients
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29 Sep 2015
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Artichoke extract
Influence of extraction temperature on Caffeoylquinic acids
Wat
er
Brand, N.: Deutsche Apotheker Zeitung · 137. Jahrgang · Nr. 41 · 9 10 1997
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29 Sep 2015
42
INSTRUÇÃO NORMATIVA Nº 4, 18 JUNHO 2014
2.9 VALIDAÇÃO DE MÉTODOS ANALÍTICOS
Revalidações
As metodologias analíticas devem ser revalidadas em caso de
mudanças significativas na obtenção ou composição da matériaprima, mudanças na composição do produto acabado ou mudanças
no procedimento analítico. Dependendo do grau de alteração
realizada, apenas uma validação parcial (incluindo seletividade,
exatidão e precisão) será suficiente. A empresa deve apresentar
argumentos técnicos que justifiquem essa medida.
H. Sievers, ANVISA Workshop Brasilia
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Summary and Conclusion
The specification does not describe/define an herbal API in its totality
because herbal API are strongly defined by their production process
starting with the cultivation/collection of plant material.
Thus, a change of the extract manufacturer is a major variation with
potential influence on quality, safety and efficacy and thus requires
thorough assessment and documentation of
quality of plant material (identity, contaminants, composition)
manufacturing process
final composition of the preparation (incl. excipients)
content of markers (and toxic compounds where applicable)
analytical methods for herbal drug and API
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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Summary and Conclusion
Exchange of the extract manufacturer may potentially affect most
parts of module 3.2.S of the dossier or the relevant chapters and
information according to INSTRUÇÃO NORMATIVA Nº 4, 18
JUNHO 2014, respectively.
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
45
Thanks to
Eliana Bufaino
Bernhard Klier
Ines Lederer
Klaus Reif
Thank you!
Your contact:
Dr. Hartwig Sievers
PhytoLab GmbH & Co. KG
Dutendorfer Straße 5-7
91487 Vestenbergsgreuth
Germany
Tel.: +49 9163 88-216.
Fax.: +49 9163 88-349
[email protected]
H. Sievers, ANVISA Workshop Brasilia
29 Sep 2015
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H. Sievers, ANVISA Workshop Brasilia
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