2.4. Bağımsız Denetim Kalitesini Etkileyen Faktörler
2.4.1. Bağımsız Denetim Kuruluşu ile İlgili Faktörler
2.4.1.2. Denetim Kuruluşunun Büyüklüğü
O presente estudo demonstrou sucesso na previsão do fator de retenção de 7
corticosteroides utilizando o algoritmo S-RLM. Os altos coeficientes de correlação e
baixíssimos erros de previsão confirmaram o alto poder de previsão do modelo
desenvolvido. Na analise simultânea desses compostos o método ideal previsto
demonstrou um bom método isocrático para 4 compostos analisados durante somente 3
minutos de eluição. A aplicação do modelo para testar uma série de compostos
demonstrou que este novo modelo é capaz de prever com excelente precisão o fator de
retenção de corticosteroides mesmo em condições extrapoladas. Espera-se aprimorar
esse simulador no sentido de atender uma variedade de compostos esteroidais.
Conclusões
113Os resultados desse trabalho mostraram que foi possível aplicar a abordagem
aQbD para o desenvolvimento de métodos cromatográficos de ultra eficiência. Esse
estudo foi focado em análise simultânea de compostos corticosteroides, antifúngicos e
também a determinação de conservantes utilizados em formulações semissólidas.
No primeiro capítulo, a identificação e quantificação de dexametasona acetato e
clotrimazol em cremes, foi realizada através de software de transferência de método de
CLAE para CLUE, e foi obtido um método mais rápido, havendo pouco gasto de
solvente durante a otimização para CLUE, além do preparo de amostra ter sido simples
e rápido. O método ofereceu vantagens em análises de rotina para essas formulações,
quando comparado aos métodos oficiais e os já desenvolvidos para CLAE.
Foi visto também que a determinação simultânea de compostos é um desafio
quando se deseja trabalhar com métodos isocráticos. Para isso, no segundo capítulo, o
método foi desenvolvido através de simulação em software comercial (ChromSword
Auto Rapid
®) com preferência e ajuste para método isocrático, uma vez que esse modo
de eluição permite uma melhor reprodutibilidade e robustez. No entanto, o software
possuiu limitação, desenvolvendo método somente em modo CLAE. A solução
encontrada, após o desenvolvimento do método pelo software, foi transferir as
condições cromatográficas para CLUE e verificar a eficiência da separação em coluna
com partícula de núcleo sólido, que exige condições de ultra pressão. Três colunas
utilizadas em CLUE foram comparadas em relação à sua eficiência, e o método mostrou
ser adequado para análise simultânea de corticosteroides e conservantes utilizando uma
coluna Core-Shell 1.3µm quando se injetou 1 µL de amostra. O método apresentou
menor tempo de análise e menor consumo de solvente, quando comparado ao método
por CLAE.
No terceiro capítulo, o planejamento fatorial completo 3
3foi utilizado como
ferramenta aQbD para desenvolvimento de métodos. O planejamento mostrou zonas de
robustez e o método foi validado, apresentando seletividade e linearidade, atendendo
principalmente aos parâmetros de adequabilidade do sistema. O método foi aplicado na
análise de creme, gel, loção e pomada contendo betametasona valerato, utilizando
dexametasona acetato como padrão interno. O tempo total por corrida e o baixo
consumo de solvente, 0,2 mL por minuto, mostrou que o método além de ser robusto,
preciso e exato, ofereceu vantagens em relação aos métodos oficiais presentes nas
Farmacopéias Americana e Britânica, sendo mais rápido, econômico e confiável.
Conclusões
114Frente às dificuldades encontradas na simulação de retenção de compostos, em
condições de ultra pressão, tornou-se necessária a criação de um modelo matemático
que pudesse atender a essas condições cromatográficas diferenciadas. No quarto
capítulo, o conceito de correlação quantitativa entre estutura química e retenção foi
utilizado para criação de um modelo de previsão de fator de retenção para análise de
corticosteroides em cromatografia líquida de ultra eficiência. O modelo desenvolvido
apresentou excelente condição isocrática para análise simultânea de corticosteroides em
menos de três minutos de corrida. A previsão foi eficiente mesmo em condições
extrapoladas de fase móvel e temperatura do forno da coluna. Esse modelo foi aplicado,
portanto, na criação de um simulador para análise desses compostos em diferentes
condições cromatográficas de ultra eficiência.
Seja com o planejamento estatístico ou uso de ferramentas computacionais, os
métodos ainda necessitam de intervenções para ajuste, otimização e tomada de decisões.
O trabalho mostrou que apesar de terem sido planejados por diferentes estratégias, a
intervenção do analista/cromatografista ainda é necessária. Tendências, como o uso de
inteligência artificial e modelos mais sofisticados, são promessas para um futuro
promissor na cromatografia e na análise de fármacos e medicamentos.
ABRAHAM, M. H.; IBRAHIM, A.; ZISSIMOS, A. M. Determination of sets of solute
descriptors from chromatographic measurements. Journal of Chromatography A, v.
1037, n. 1-2, p. 29–47, 2004.
ALAN XU, Q. Ultra-High Performance Liquid Chromatography and Its
Applications. New Jersey: John Wiley & Sons, 2013.
AMBURE, P. et al. “NanoBRIDGES” software: Open access tools to perform QSAR
and nano-QSAR modeling. Chemometrics and Intelligent Laboratory Systems, v.
147, p. 1–13, 2015.
AUTHORS, U.; GUILLARME, D.; VEUTHEY, J. Guidelines for the use of UHPLC
Instruments. p. 1–11, [s.d.].
BACZEK, T. et al. Comparative characteristics of HPLC columns based on quantitative
structure-retention relationships (QSRR) and hydrophobic-subtraction model. Journal
of Chromatography A, v. 1075, n. 1-2, p. 109–115, 2005.
BAJAJ, M.; NANDA, S. Full Length Research Article ANALYTICAL QUALITY BY
DESIGN ( AQBD ): NEW PARADIGM FOR ANALYTICAL METHOD
DEVELOPMENT * Mohini Bajaj and Sanju Nanda. v. 5, p. 3589–3599, 2015.
BENNETT, T. et al. Quality by Design for Analytical Methods : Implications for
Method Validation and Transfer. 2012.
BEZERRA, M.A.; SANTELLI, R.E.; OLIVEIRA, E.P.; VILLAR, L.S.; ESCALEIRA,
L. A. Response surface methodology (RSM) as a tool for optimization in analytical
chemistry. Talanta, 2008.
BOUABIDI,A.; TALBI, M.; BOUKLOUZE, A.; EL KARBANE, M.; BOURICHI, H.;
EL GUEZZAR, M.; ZIEMONS, E.; HUBERT, P.; ROZET, E. Do placebo based
validation standards mimic real batch products behaviour? Case studies. Journal of
Pharmaceutical and Biomedical Analysis, 2011.
BOUSSÈS, C. et al. Journal of Pharmaceutical and Biomedical Analysis Using an
innovative combination of quality-by-design and green analytical chemistry approaches
for the development of a stability indicating UHPLC method in pharmaceutical
products. Journal of Pharmaceutical and Biomedical Analysis, v. 115, p. 114–122,
2015.
PharmacopoeiaLondonStationery Office Books, , 2011.
BUNDGAARD, H.; HANSEN, J. Studies on the stability of corticosteroids VI. Kinetics
of the rearrangement of betamethasone-17-valerate to the 21-valerate ester in aqueous
solution. International Journal of Pharmaceutics, v. 7, n. 3, p. 197–203, 1981.
BYRNE, J.; VELASCO-TORRIJOS, T.; REINHARDT, R. Development and validation
of a novel stability-indicating HPLC method for the simultaneous assay of
betamethasone-17-valerate, fusidic acid, potassium sorbate, methylparaben and
propylparaben in a topical cream preparation. Journal of pharmaceutical and
biomedical analysis, v. 96, p. 111–7, 2014.
CARLOS, G.; URIBE, P.; FERNÁNDEZ-PEÑAS, P. Rational use of topical
corticosteroids. v. 36, n. 5, p. 158–161, 2013.
CARLUCCI, G. et al. Investigation of retention behaviour of non-steroidal anti-
inflammatory drugs in high-performance liquid chromatography by using quantitative
structure-retention relationships. Analytica chimica acta, v. 601, n. 1, p. 68–76, 2007.
CHAMSEDDIN, C.; MOLNÁR, I.; JIRA, T. Intergroup cross-comparison for the
evaluation of data-interchangeability from various chromatographic tests. Journal of
Chromatography A, v. 1297, p. 146–156, 2013.
DE CLERCQ, N.; VANDEN BUSSCHE, J.; CROUBELS, S.; DELAHAUT, P.;
VANHAECKE, L. Development and validation of a high-resolution mass-
spectrometry–based method to study the long-term stability of natural and synthetic
glucocorticoids in faeces. Journal of Chromatography A, v. 1336, 2014.
DEBRUS, B.; GUILLARME, D.; RUDAZ, S. Journal of Pharmaceutical and
Biomedical Analysis Improved quality-by-design compliant methodology for method
development in reversed-phase liquid chromatography. Journal of Pharmaceutical
and Biomedical Analysis, v. 84, p. 215–223, 2013.
DECEUNINCK, Y.; BICHON, E.; MONTEAU, F.; ANTIGNAC, J.P.; LE BIZEC, B.
Determination of MRL regulated corticosteroids in liver from various species using
ultra high performance liquid chromatography–tandem mass spectrometry (UHPLC).
No Title. Analytica Chimica Acta, v. 70, 2011.
DECEUNINCK, Y.; BICHON, E.; MONTEAU, F.; DERVILLY-PINEL, G.;
ANTIGNAC, J.P.; LE BIZEC, B. Fast and multiresidue determination of twenty
glucocorticoids in bovine milk using ultra high performance liquid chromatography–
tandem mass spectrometry. Journal of Chromatography A, v. 1294, 2013.
DECONICK, E.; COURSELLE, DE BEER, J. . Chromatography in the Detection and
Characterization of Illegal Pharmaceutical Preparations. Journal of chromatographic
science, v. 51, 2013.
DESMEDT, B.; VAN HOECK, E.; ROGIERS, V.; COURSELLE, P.; DE BEER, J.O.;
DE PAEPE, K.; DECONINCK, E. Characterization of suspected illegal skin whitening
cosmetics. Journal of Pharmaceutical and Biomedical Analysis, v. 90, 2014.
DOLAN, J. W. Gradient Elution, Part IV: Dwell-Volume Problems. LCGC North
America, v. 31, n. 6, p. 456–463, 2013.
DONG, P. P. et al. Quantitative structure-retention relationship studies for taxanes
including epimers and isomeric metabolites in ultra fast liquid chromatography.
Journal of Chromatography A, v. 1216, n. 42, p. 7055–7062, 2009.
ESMAEILPOOR, S. et al. QSRR models of veterinary drugs in milk in ultra-
performance liquid chromatography coupled to time of flight mass spectrometry. v. 98,
n. 841, p. 283–300, 2014.
FDA. Guidance For Industry. Disponível em: <www.fda.gov >.
FEDERAL, U.; CATARINA, D. E. S. CROMATOGRÁFICA ( QSRR )
EMPREGANDO DIFERENTES DESCRITORES. 2000.
FEKETE, S. et al. Trends in Analytical Chemistry Current and future trends in UHPLC.
v. 63, p. 2–13, 2014.
FU, Q. et al. Development and validation of a stability-indicating RP-HPLC method for
assay of betamethasone and estimation of its related compounds. Journal of
Pharmaceutical and Biomedical Analysis, v. 51, n. 3, p. 617–625, 2010a.
FU, Q. et al. Journal of Pharmaceutical and Biomedical Analysis Development and
validation of a stability-indicating RP-HPLC method for assay of betamethasone and
estimation of its related compounds. v. 51, p. 617–625, 2010b.
GANGADASU, B. R.; G, N. R.; DHANALAKSHMI, K. Comparison of UPLC with
UFLC : Liquid Chromatography. v. 31, n. 20, p. 97–101, 2015.
VCH, 2006.
GEDEON, C. W. The changing face of pharmaceutical analysis. v. 26, n. 1, 2007.
GIDDINGS, J. C. Unified Separation Sciences. New York: Wiley, 1991.
GOLUBOVI, J. B. et al. Quantitative structure retention relationship modeling in liquid
chromatography method for separation of candesartan cilexetil and its degradation
products. Chemometrics and Intelligent Laboratory Systems, v. 140, p. 92–101,
2015.
GONZÁLEZ-RUIZ, V.; OLIVES, A. I.; MARTÍN, M. A. Trends in Analytical
Chemistry Core-shell particles lead the way to renewing high-performance liquid
chromatography. Trends in Analytical Chemistry, v. 64, p. 17–28, 2015.
GORYŃSKI, K. et al. Quantitative structure–retention relationships models for
prediction of high performance liquid chromatography retention time of small
molecules: Endogenous metabolites and banned compounds. Analytica Chimica Acta,
v. 797, p. 13–19, 2013.
GOSETTI, F.; MAZZUCCO, E.; GENNARO, M.C.; MARENGO, E. Ultra high
performance liquid chromatography tandem mass spectrometry determination and
profiling of prohibited steroids in human biological matrices. Journal of
Chromatography B, v. 927, 2013.
GRITTI, F.; GUIOCHON, G. Repeatability of the efficiency of columns packed with
sub-3 m core – shell. Journal of Chromatography A, v. 1252, p. 45–55, 2012.
GRITTI, F.; GUIOCHON, G. core – shell particles. Journal of Chromatography A, v.
1280, p. 35–50, 2013.
GUILLARME, D.; VEUTHEY, J. L. UHPLC in Life Sciences. Cambridge: Royal
Society of Chemistry, 2012.
GUILLARME, D. et al. Recent developments in liquid chromatography — Impact on
qualitative and quantitative performance. v. 1149, p. 20–29, 2007.
GUILLARME, D. et al. Method transfer for fast liquid chromatography in
pharmaceutical analysis : Application to short columns packed with small particle . Part
II : Gradient experiments. v. 68, p. 430–440, 2008.
Biomedical Analysis Reliability of computer-assisted method transfer between several
column dimensions packed with 1 . 3 – 5 m core – shell particles and between
various instruments. v. 94, p. 188–195, 2014.
HE, C.; FAN, H.; TAN, J.; ZOU, J.; ZHU, Y.; YANG, K.; HU, Q. Pharmacokinetics of
betamethasone and betamethasone-17-monopropionate in Chinese healthy volunteers
after intramuscular injection of betamethasone phosphate/ betamethasone dipropionate.
Arznei-forschung, v. 61, 2011.
HEATON, J. C.; MCCALLEY, D. V. Comparison of the kinetic performance and
retentivity of sub-2 m core – shell , hybrid and conventional bare silica phases in
hydrophilic interaction chromatography. Journal of Chromatography A, v. 1371, p.
106–116, 2014.
HERRERO, P.; BORRULL, F.; MARCÉ, RM.; POCURULL, E. Pressurised liquid
extraction and ultra-high performance liquid chromatography-tandem mass
spectrometry to determine endogenous and synthetic glucocorticoids in sewage sludge.
Talanta, v. 103, 2013.
HERRERO, P.; BORRULL, F.; POCURULL, E.; MARCÉ, R. M. Determination of
glucocorticoids in sewage and river waters by ultra-high performance liquid
chromatography–tandem mass spectrometry. Journal of Chromatography A, v. 1224,
2012.
HEWITT, E. F.; LUKULAY, P.; GALUSHKO, S. Implementation of a rapid and
automated high performance liquid chromatography method development strategy for
pharmaceutical drug candidates. v. 1107, p. 79–87, 2006.
HIBBERT, D. B. Experimental design in chromatography: A tutorial review. Journal
of Chromatography B, v. 910, p. 2–13, 2012.
HOANG, T. H. et al. Short communication C omputer-assisted method development
and optimization in high-performance liquid chromatography. v. 991, p. 281–287, 2003.
ICH. ICH International Conference on Harmonisation of Technical Requirements
for Registration of Pharmaceuticals for Human Use. Q2(R1)Validation of
Analytical Procedures.Geneva, 1996.
IRWIN, J. J. et al. ZINC: a free tool to discover chemistry for biology. Journal of
JACKSON, P. Quality by Design in Pharmaceutical Analysis. n. October, 2015.
JINNO, K. Chromatographic Separations Based on Molecular Recognition. New
York: Wiley-VCH, 1997.
JOSEPH MOSES JURAN. A qualidade desde o projeto: novos passos para o
planejamento da qualidade em produtos e serviços. [s.l.] Pioneira, 1994.
KALISZAN, R. Quantitative Structure Chromatographic Retention Relationships.
New York: Wiley, 1987.
KALISZAN, R. Structure and Retention in Chromatography. A Chemometric
Approach. Amsterdam: Harwood Academic Publishers, 1997.
KALISZAN, R. Recent advances in quantitative structure-retention relationships (
QSRR ). v. 1, n. 11 1, p. 530–534, 2000.
KALISZAN, R. Quantitative structure –( chromatographic ) retention relationships.
Chemical Reviews, v. 107, p. 3212–3246, 2007.
KALISZAN, R.; HARTWICK, R. A. Quantitative Relationship Between Molecular
Structure and Chromatographic Retention. Implications in Physical, Analytical, and
Medicinal Chemistry. C R C Critical Reviews in Analytical Chemistry, v. 16, n. 4, p.
323–383, 1986.
KARBANE, M. EL et al. Development and validation of a reversed-phase HPLC
method for simultaneous analysis of butylhydroxyanisol , simvastatin and its. Annales
Pharmaceutiques Francaises, v. 72, n. 4, p. 244–255, 2014.
KAUTSKY, M. B. Steroid Analysis by HPLC. New York: Marcel Dekker, 1981.
LIN, M.; WU, N. Comparison between micellar electrokinetic chromatography and
HPLC for the determination of Betamethasone Dipropionate , Clotrimazole and their
related substances. v. 19, p. 945–954, 1999.
LUNN, G. HPLC Methods for Recently Approved Pharmaceuticals. New Jersey:
Willey & Sons, 2005.
MALDANER, L.; CRISTINA, I.; FONTES, S. UHPLC – Uma abordagem atual :
desenvolvimentos e desafios recentes. v. 4, n. 3, p. 197–207, 2012.
MARKUSZEWSKI, M.; KALISZAN, R. Quantitative structure-retention relationships
in affinity high-performance liquid chromatography. Journal of Chromatography B:
Analytical Technologies in the Biomedical and Life Sciences, v. 768, n. 1, p. 55–66,
2002.
MASON, R.; GUNST, R.F.; HESS, J. . Statistical Design and Analysis of
Experiments. 2nd. ed. New York: Wiley-Interscience, 2003.
MAURI, A et al. Dragon software: An easy approach to molecular descriptor
calculations. Match Communications In Mathematical And In Computer
Chemistry, v. 56, n. 2, p. 237–248, 2006.
MCCALLEY, D. V. Some practical comparisons of the efficiency and overloading
behaviour of sub-2 m porous and sub-3 m shell particles in reversed-phase liquid
chromatography. Journal of Chromatography A, v. 1218, n. 20, p. 2887–2897, 2011.
MYERS, R.H.; MONTGOMERY, D. . Response Surface Methodology: Process and
Product Optimization Using Designed Experiments. 2nd. ed. New York: Wiley,
2002.
NOORIZADEH, H.; FARMANY, A.; NOORIZADEH, M. Quantitative structure-
retention relationships analysis of retention index of essential oils. Quimica Nova, v.
34, n. 2, p. 242–249, 2011.
NOORIZADEH, H.; NOORIZADEH, M.; FARMANY, A. Advanced QSRR models of
toxicological screening of basic drugs in whole blood by UPLC-TOF–MS. Medicinal
Chemistry Research, v. 21, n. 12, p. 4357–4368, 2012.
NOORIZADEH, H.; NOORIZADEH, M.; MUMTAZ, A. S. {QSRR} analysis of
capacity factor of nanoparticle compounds. Journal of Saudi Chemical Society, v. 18,
n. 3, p. 183–189, 2014.
NOV, L.; MATYSOV, L.; SOLICH, P. Advantages of application of UPLC in
pharmaceutical analysis. v. 68, p. 908–918, 2006.
NOVÁKOVÁ, L.; MATYSOVÁ, L.; SOLICH, P. Advantages of application of UPLC
in pharmaceutical analysis. Talanta, v. 68, 2006.
ORLANDINI, S.; PINZAUTI, S.; FURLANETTO, S. Application of quality by design
to the development of analytical separation methods. p. 443–450, 2013.
PARKER, J. PARKER, P. Betamethasone – A medical dictionary, bibliography and
Publications, 2004.
PEÑA-BRIOLES, D.; GONZALO-LUMBRERAS, R.; IZQUIERDO-HORNILLOS,
R.; SANTOS-MONTES, A. Method development for betamethasone and
dexamethasone by micellar liquid chromatography using cetyl trimethyl ammonium
bromide and validation in tablets. Journal of Pharmaceutical and Biomedical
Analysis, v. 36, 2004.
PERAMAN, R.; BHADRAYA, K.; REDDY, Y. P. Analytical Quality by Design : A
Tool for Regulatory Flexibility and Robust Analytics. v. 2015, 2015.
PLENIS, A. Comparison of core – shell and totally porous ultra high performance liquid
chromatographic stationary phases based on their selectivity towards alfuzosin
compounds ଝ. v. 1346, p. 69–77, 2014.
PLENIS, A.; OLEDZKA, I.; BACZEK, T. Classification of LC columns based on the
QSRR method and selectivity toward moclobemide and its metabolites. Journal of
Pharmaceutical and Biomedical Analysis, v. 78-79, p. 161–169, 2013.
POOLE, C. . The Essence of Chromatography. Amsterdam: Elsevier Science, 2003.
R DEVELOPMENT CORE TEAM. R: A Language and Environment for Statistical
Computing. R Foundation for Statistical ComputingVienaR Foundation for
Statistical Computing, , 2011.
RAMAMOORTHY, S.; CIDLOWSKI, J. A. Corticosteroids. Rheumatic Disease
Clinics of North America, v. 42, n. 1, p. 15–31, 2016.
RAMAN, N. V. V. S. S.; MALLU, U. R.; BAPATU, H. R. Analytical Quality by
Design Approach to Test Method Development and Validation in Drug Substance
Manufacturing. v. 2015, 2015.
Random. Disponível em: <random.org>. Acesso em: 1 jan. 2015.
ROZET, E. et al. Quality by Design Compliant Analytical Method Validation. p. 106–
112, 2012.
RUTA, J.;CABOOTER, D.; RUDAZ, S.; GERT DESMET, VEUTHEY, J.L.;
GUILLARME, D. Method development for pharmaceutics: Some solutions for tuning
selectivity in reversed phase and hydrophilic interaction liquid chromatography.
RUTA, J. et al. Evaluation of columns packed with shell particles with compounds of
pharmaceutical interest. Journal of Chromatography A, v. 1228, p. 221–231, 2012.
SCHÄCKE, H.; DÖCKE, W. D.; ASADULLAH, K. Mechanisms involved in the side
effects of glucocorticoids. Pharmacology & therapeutics, v. 96, n. 1, p. 23–43, 2002.
SCHMIDT, A. H.; MOLNÁR, I. Journal of Pharmaceutical and Biomedical Analysis
Using an innovative Quality-by-Design approach for development of a stability
indicating UHPLC method for ebastine in the API and pharmaceutical formulations.
Journal of Pharmaceutical and Biomedical Analysis, v. 78-79, p. 65–74, 2013.
SEIFERT, E.; ABRAMO, L.; THELIN, B. Experimental design and optimization. 1998.
SMITH, E. W.; HAIGH, J. M.; KANFER, I. A stability-indicating HPLC assay with on-
line clean-up for betamethasone 17-valerate in topical dosage forms. International
Journal of Pharmaceutics, v. 27, n. 2-3, p. 185–192, 1985.
SNYDER, L.R.; KIRKLAND, J.J.; DOLAN, J. W. Introduction to Modern Liquid
Chromatography. 3rd. ed. New Jersey: Wiley, 2010.
TAKEMURA, C. et al. Effect of Temperature on Retention of Diazines in.pdf. v. 34, n.
2, 2013.
TAVARES, L. C. QSAR: A abordagem de Hansch. Quimica Nova, v. 27, n. 4, p. 631–
639, 2004.
The International Conference on Harmonisation of Technical Requirements for
Registration of Pharmaceuticals for Human Use (ICH). Disponível em:
<www.ich.org>.
THYE, B.; SOARES, M.; KUMAR, A. Méritos comparativos da Cromatografia em
Fase Líquida de Alta Eficiência em escala convencional e minituarizada. v. 92, n. 2, p.
44–50, 2011.
TICHÝ, M.; RUCKI, M. Validation of QSAR models for legislative purposes.
Interdisciplinary toxicology, v. 2, n. 3, p. 184–6, 2009.
USP. United States Pharmacopoeia USP 34 NF 29USARockville, , 2011.
VOGT, F. G.; KORD, A. S. Development of Quality-By-Design Analytical Methods. v.
100, n. 3, p. 797–812, 2011.
and HPLC Method Upgrade : Importance of Selectivity and Efficiency. [s.d.].
WATERS. Ultra Performance LC by designUSA, 2004.
WREN, S. A. C. Peak capacity in gradient ultra performance liquid chromatography (
UPLC ). v. 38, p. 337–343, 2005.
WU, C.F.J.; HAMADA, M. Experiments: Planning, Analysis, and Parameter
Design Optimization. [s.l.] Wiley, 2000.
XIAO, K. P. et al. Efficient method development strategy for challenging separation of
pharmaceutical molecules using advanced chromatographic technologies. v. 1163, p.
145–156, 2007.
XIONG, Y.; XIAO, K. P.; RUSTUM, A. M. Development and validation of a stability-
indicating RP-HPLC method to separate low levels of dexamethasone and other related
compounds from betamethasone. Journal of Pharmaceutical and Biomedical
Analysis, v. 49, n. 3, p. 646–654, 2009.
YAO, H. et al. Journal of Pharmaceutical and Biomedical Analysis An analytical
quality by design ( aQbD ) approach for a l -asparaginase activity method. Journal of
Pharmaceutical and Biomedical Analysis, v. 117, p. 232–239, 2016.
YOGO, K. et al. Prediction of Chromatographic Retention of Pyrazine and
Alkylpyrazines in RP-LC. Chromatographia, v. 70, n. 5, p. 677–684, 2009.
YOSHIE, P.; DE, F. C. METODOLOGIA ANALÍTICA DE CITRATO DE
SILDENAFIL. [s.d.].
YU JIN, XINGYA XUE, HUI SHI, YUANSHENG XIAO, FEIFANG ZHANG, X. L.
HPLC and UPLC Switch for TCM analysis. v. 10, n. 1, p. 80–84, 2008.
YU, L. X. et al. Review Article Understanding Pharmaceutical Quality by Design. v.
16, n. 4, 2014.
ZALEWSKI, P.; TALACZYN, A. An Approach to Transfer Methods from HPLC to
UHPLC Techniques in Some Carbapenems. p. 1483–1487, 2014.
Anexo I
Estrutura das moléculas utilizadas no
estudo
Testesterone propionate B-estradiol Prednisone Betamethasone valerate Dexamethasone acetate Desonide Hydrocortisone acetate Clobetasol propionate Fluocinolone acetonide
Anexo II
Development and validation of an UHPLC method for the determination
of betamethasone valerate in cream, gel, ointment and lotion
Lílian Grace da Silva Solona, Igor Prado de Barros Limaa, Fernando Henrique Andrade Nogueiraa,
Jailton Paulo de Araújob, Carla Almeida Vivacquab, Cícero Flávio Soares Aragãoa,⇑
aDepartamento de Farmácia, Universidade Federal do Rio Grande do Norte, Laboratório de Controle de Qualidade de Medicamentos, Rua General Gustavo Cordeiro de Faria, S/N, 59012-570 Natal, RN, Brazil
bDepartamento de Estatística, Laboratório de Estatística Aplicada, Universidade Federal do Rio Grande do Norte, Caixa Postal 1524, Campus Universitário Lagoa Nova, 59078-970 Natal, RN, Brazil
a r t i c l e i n f o
Article history:
Received 24 September 2015
Received in revised form 19 November 2015 Accepted 16 December 2015
Available online 18 December 2015
Keywords:
Betamethasone valerate Dermatologic formulations DoE
Three-level factorial design UHPLC
a b s t r a c t
An ultra high performance liquid chromatographic method has been developed and validated for the determination of betamethasone valerate (BMV) in topical dermatologic formulations. For the develop- ment of the method, response surface methodology based on a three-level full factorial design was used. The eluent composition, the column dimension and the flow rate were chosen as relevant experimental parameters to investigate. The response surface plots revealed an optimum separation by using a RP col-
umn (30 mm 2 mm i.d., 2.2lm particle size), at 30 °C; isocratic mobile phase consisting of acetonitrile:
water (60:40) at a flow rate of 0.2 mL min 1and a wavelength set at 254 nm. The proposed method was
validated for four types of matrices according to ICH guidelines requirements. Dexamethasone acetate
(DMA) was used as internal standard. Linearity was studied in the range of 5–200lg mL 1for BMV in
spiked matrix samples. Recoveries were in the range of 95–105% and precision was better than 5% for both analytes, either in cream, gel, ointment, or lotion formulations, when using simple sample prepara- tion. Retention times were 0.95 min for DMA and 1.40 min for BMV, demonstrating a short method run time. The method was successfully applied for routine analysis of dermatological formulations containing