Programa de Pós-Graduação em Ciências Farmacêuticas

Mestrado Profissional em Ciência e Tecnologia Farmacêutica

Caracterização de Sistemas Particulados
Prof Dr. Luiz Cláudio R. Pereira da Silva

Introdução à Caracterização de Sistemas

Particulados

06/04/2015
Conceituação de Sistemas Particulados em Ciências
Farmacêuticas

Micro

Nano
 Conceituação de Sistemas Particulados em Ciências
Farmacêuticas
O que é uma partícula?

Discreta subporção de uma substância.

Para estreitar a definição ao propósito da disciplina dizemos que são partículas

sólidas ou gotículas líquidas com dimensões físicas entre a faixa subnanométrica
e alguns milímetros de tamanho.

Os tipos de materiais mais comuns que compõem partículas são:

Pós;
Grânulos;
Suspensões;
Emulsões;
Classificação de Sistemas Particulados
Relação de características estudadas
Relação de características estudadas
Relação de características estudadas
Relação de características estudadas
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
 Motivações para Caracterização de Sistemas Particulados

Transposição de barreiras  epitélios intestinal, pulmonar,

cutâneo; endotélio vascular; barreira mucosa;

Interações com sistemas biológicos  trombina; EGFR; VEGFR;

Direcionamento  BHE; células tumorais; inflamação;

coagulação

Agregação  meio biológico; agente dispersante; pH

Toxicidade  SNC; ROS; Inalação

Degradação  PLGA; PCL; Quitosana; albumina; lipídios; ouro,

prata; etc...
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
Desafios no Processo de Caracterização de Sistemas
Particulados

Ausência de
harmonização
Validação de técnicas
especializada

Histórico Regulação
recente das ausente ou
técnicas primária

Complexidade
físico-química
 Desafios no Processo de Caracterização de Sistemas
Particulados

Physicochemical properties:

Traditional small molecule drugs are characterized by their molecular weight, chemical
composition, purity, solubility, and stability. These data form the basis of the chemistry,
manufacturing, and controls. For small molecules, the instrumentation to ascertain these
properties have been well established and the techniques are standardized. Techniques like
nuclear magnetic resonance (NMR), mass spectrometry (MS), ultraviolet-visible (UV-Vis)
spectroscopy, infrared spectroscopy (IR), and gas chromatography (GC) can be run in a high-
throughput fashion to analyze such molecules.

For nanomaterials, alternate instrumentation is required to obtain information on the same

properties (composition, purity, stability, etc.). These properties influence biological activity,
and may depend on parameters such as particle size, size distribution, surface area, surface
charge, surface functionality, shape, and aggregation state.

Additionally, since many nanoparticle concepts are multifunctional (with targeting, imaging,
and therapeutic components), the stability and distribution of these components can have
dramatic effects on nanoparticle biological activity as well.
 Desafios no Processo de Caracterização de Sistemas
Particulados

It is now widely acknowledged that physicochemical properties such as size and surface
chemistry can dramatically affect nanoparticle behavior in biological systems and influence
biodistribution, safety and efficacy. For instance, a decrease in particle size leads to an
exponential increase in surface area per unit mass, and a concomitant increase in the
availability of reactive surface groups.

Nanoparticles with cationic surfaces have a notably increased tendency to permeate (and
perforate) cellular membranes compared to neutral or anionic nanoparticles.
Physicochemical characterization of properties such as size, surface area, surface chemistry,
and aggregation state can provide the basis for better understanding of structure–activity
relationships.

Methods are presented for determining nanoparticle size in solution by dynamic light
scattering (DLS), molecular weight via mass spectrometry, surface charge through zeta
potential measurement and topology by atomic force microscopy (AFM). Methods are also
presented for transmission electron microscopy (TEM) and scanning electron microscopy
(SEM) examination of nanoparticle samples, and elemental identification using energy
dispersive X-ray spectroscopy (EDX).
 Desafios no Processo de Caracterização de Sistemas
Particulados

Another important and challenging area of nanoparticle characterization is

measurement under physiological conditions that resemble or mimic the physical state
in vivo.

Many properties of nanoparticles are environment and condition dependent; for

example:
Particle’s hydrodynamic size at physiological pH and ionic strength may differ from the
size in water or the dry state;
Surface charge may also depend on the pH and ionic strength of the suspending
solution;
Plasma proteins are known to bind nanoparticles in the blood, and the protein-bound
size is expected to be a more relevant determinant of disposition and clearance than the
free-particle size;
The release profile of an encapsulated therapeutic may even be environment
dependent. That is, there may be faster or slower release of the therapeutic as the
temperature, pH, and/or ionic strength of the solution surrounding the nanoparticle
formulation is varied.
 Desafios no Processo de Caracterização de Sistemas
Particulados
Because the results of in vitro biological assays often In vitro studies may also be used
don’t correlate with in vivo endpoints, in vitro to identify areas requiring
characterization is performed to elucidate mechanisms, attention for in vivo animal
not necessarily to screen for biocompatibility. studies.
Unfortunately, nanoparticle-
based therapeutics frequently
interfere with conventional in
vitro pharmacologic assays. For
instance, many nanoparticles
aggregate or adsorb proteins.
Other nanoparticles scatter light
or have optical properties, which
may invalidate colorimetric
assays that rely on absorbance
measurements. Some
nanoparticles have catalytic
properties that may interfere
with enzymatic tests, such as
those that evaluate endotoxin
contamination.
http://ncl.cancer.gov/working_assay-cascade.asp
http://ncl.cancer.gov/working_assay-cascade.asp
http://ncl.cancer.gov/working_assay-cascade.asp
Numerous reviews have been published focusing on the state of knowledge of
behavior, interaction with biological systems (both for advantageous and toxicity
outcomes), and potential environmental disposition of nanoscale materials.
Government bodies have published some of these, as have private entities
focusing on the risk associated with nanoscale materials, and still others can be
found in the peer-reviewed public literature. Often these reports stress the need
for research on: characterization of materials in the nanoscale range; methods for
identifying hazards; understanding biological response to nanoscale materials;
and characterizing nanoscale material exposure and transport (in humans and the
environment).

In light of the current state of the science, an understanding of the interactions of

nanoscale materials with biological systems is probably best gained through case-
by-case analysis of specific types of nanoscale materials and of variations in their
characteristics.
Generalizable principles are being derived from studies of specific types of
nanoscale materials. For instance, studies in some laboratories have determined
that surface area per unit of volume or mass can be a better measure than mass
for assessing relative toxicity across particle size and material variations. This tells
us, for example, that the suitability of mass-based dose measurements should be
evaluated before drawing conclusions about potency of a drug effect or about toxic
response. However, it is important to recognize that in some cases surface area
has been shown to be a much less important determinant of biological interaction
than surface modification (for example, binding different chemical groups to the
surface of a particle) for those particular types of particles in which this has been
examined systematically. This indicates that one should pay particular attention to
the composition and surface characteristics of nanoscale materials that may come
in contact with biological systems.
Comments were directed to the inadequacy or absence of currently available and
standardized product characterization methodologies for nanoscale materials. A
comment highlighted that FDA has limited ability to detect nanoscale material
components in some products. Another comment emphasized that FDA's ability to
inspect products is also presently significantly limited with regard to products that
may contain nanoscale materials. One comment recommended that nanoscale
material be characterized with respect to size (surface area and size distribution),
chemical composition (such as purity and crystallinity), surface structure (surface
reactivity, surface groups, inorganic/organic coatings, etc.), solubility, shape and
aggregation. Other comments noted that detection of nanoscale materials requires
expensive and sophisticated equipment, and it is often unclear which parameters are
relevant to toxicity.
Comments also encouraged FDA to work with other government agencies to
develop rapid screening tools for these types of products. Several comments noted
that the Nanotechnology Characterization Laboratory, run by the National Cancer
Institute, will be very useful in helping to characterize nanoscale materials and to
develop standards and standardized methods for measuring nanoscale materials.
Metrology - Characterization of Particles in the Nanoscale Range
Currently, ability to detect nanoscale materials in the body or in products
regulated by FDA is limited, and development of appropriate analytical methods
for classes of products and of nanoscale materials may require substantial effort.
Further, new analytical methods, and methods that FDA reviewers are generally
less familiar with, are often used to characterize nanoscale materials. The
strengths and limitations of these methods may vary in ways relevant to
evaluating characteristics such as particle size, size distribution, surface charge,
surface properties, and particle interactions (such as aggregation) that may be
relevant to dose, stability, or other characteristics significant to biological
interaction or product quality.
Standardization of Tests and Data Reporting
In many cases, methods currently used to characterize nanoscale materials
have not been standardized through recognized standard development
organizations. There is a need to develop standard particle characterization
methods. Furthermore, given the range of methods being used and being
developed for nanoscale materials, there is a need to develop consistent
nomenclature and measurement types and formats for use across studies and
data submissions similar to the "MIAME" approach developed for microarray
data. Consistent reporting approaches will make data from one laboratory or for
one material type more readily comparable to data from other laboratories and
more useful to subsequent consideration of properties of the same type of
material or other material types.
A rational characterization
In vitro
strategy for biomedical
nanoparticles contains three
elements: physicochemical
characterization, in vitro assays,
and in vivo studies.

Fisico-
químic
o

In vivo
luizclaudio@pharma.ufrj.br