Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles
U susret pravilnom određivanju veličine nanostruktura: analiza uticaja uslova merenja i pripreme uzorka na razultate procene veličine nanokapi i nanočestica
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Physicochemical properties of many active ingredients jeopardize their
pharmacological activity. To overcome identified obstacles, nanosystems as carriers for
delivery of actives have been recognized as promising tools, with highly posted expectations.
The nanotechnology-based approach in drug formulation is much more than just another
step in size miniaturization. Given the complexity of nanosystems, challenges encountered in
their characterization, and evident lack of testing protocols, relevant European
research/regulatory bodies have issued guidelines, summarizing important parameters for
nanosystem characterization. Size/size distribution (per se and in biorelevant environment)
are essentially important, representing critical quality attributes (1). The aim was to show
how significantly different results could be obtained under different measurement
conditions/sample preparation methods, offering optimal protocol for size estimation
applying dynamic light scattering (DL...S), with complementary analysis through atomic force
microscopy. Hydrophilic nanoemulsion and aqueous dispersion of polymeric nanoparticles
were used as test samples. Ultrapurified water, phosphate buffer saline (PBS, pH 7.4) and
biorelevant medium with serum proteins were used for dilution. Measurements were
performed applying batch-mode DLS (ZetasizerNano), and AutoProbe CP-Research
microscope. Significant differences in obtained nanodroplet/nanoparticle size were
observed depending on the type of medium and dilution level. Protein corona formation
could not be confirmed with certainty. Preference was given to PBS as dispersant. The
optimal level of dilution for nanoparticles was 1:10 (Z‐ave=59.16±0.46nm), and for
nanoemulsion 1:100 (Z‐ave=73.5±0.75nm). For proper interpretation, it is necessary that the
DLS measurement report, in addition to the Z‐ave and the polydispersity index, contains at
least the data on the attenuation and correlation function intercept.
Fizičkohemijske osobine mnogih aktivnih supstanci otežavaju ostvarivanje
farmakoloških efekata. Kako bi se identifikovane prepreke prevazišle, visoka očekivanja
postavljena su pred nanosisteme - nosače za isporuku aktivnih supstanci, uz značajna
ulaganja u njihov razvoj. Nanotehnološki pristup formulaciji leka mnogo je više od pukog
smanjenja veličine. Imajući u vidu kompleksnost nanosistema, izazove koji se sreću u
njihovoj karakterizaciji, te evidentan nedostatak protokola ispitivanja, relevanta evropska
istraživačka/regulatorna tela izdala su vodiče, sumirajući značajne parametre karakterizacije
nanosistema. Veličina/distribucija veličina (per se i u biorelevantnim uslovima) od suštinske
su važnosti za postizanje očekivanih performansi, predstavljajući kritične atribute kvaliteta
(1). Cilj rada bio je pokazati kako se pri različitim uslovima merenja/načinima pripreme
istog uzorka dobijaju značajno različiti rezultati procene veličine, te ponuditi optimalan
protokol merenj...a dinamičkim rasipanjem svetlosti (DLS), uz komplementarno ispitivanje
mikroskopijom atomskih sila. Kao test uzorci korišćeni su hidrofilna nanoemulzija i vodena
disperzija polimernih nanočestica, a kao medijumi za razblaživanje visokoprečišćena voda,
izotonični fosfatni pufer (PBS, pH 7,4) i biorelevantni medijum obogaćen proteinima seruma.
Merenja su sprovedena primenom batch‐mode DLS, (ZetasizerNano), i na mikroskopu
AutoProbe CP-Research. Primećene su značajne razlike u vrednostima veličine
nanokapi/nanočestica zavisno od tipa medijuma i nivoa razblaženja. Nije se moglo sa
sigurnošću potvrditi formiranje protein korone nakon inkubacije u biorelevantnom
medijumu. Za procenu veličine per se, prednost je data PBS-u kao disperzantu. Optimalan
nivo razblaženja za nanočestice bio je 1:10 (Z‐ave=59,16±0,46nm), a za nanoemulziju 1:100
(Z‐ave=73,5±0,75nm). Radi adekvatnog tumačenja, potrebno je da izveštaj DLS merenja,
pored Z-ave i indeksa polidisperznosti, sadrži barem još podatke o atenuaciji i odsečku
korelacione funkcije.
Source:
Arhiv za farmaciju, 2022, 72, 4 suplement, S192-S193Publisher:
- Savez farmaceutskih udruženja Srbije (SFUS)
Funding / projects:
Note:
- VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beograd
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Institution/Community
PharmacyTY - CONF AU - Nikolić, Ines AU - Ranđelović, Danijela AU - Savić, Snežana PY - 2022 UR - https://farfar.pharmacy.bg.ac.rs/handle/123456789/4493 AB - Physicochemical properties of many active ingredients jeopardize their pharmacological activity. To overcome identified obstacles, nanosystems as carriers for delivery of actives have been recognized as promising tools, with highly posted expectations. The nanotechnology-based approach in drug formulation is much more than just another step in size miniaturization. Given the complexity of nanosystems, challenges encountered in their characterization, and evident lack of testing protocols, relevant European research/regulatory bodies have issued guidelines, summarizing important parameters for nanosystem characterization. Size/size distribution (per se and in biorelevant environment) are essentially important, representing critical quality attributes (1). The aim was to show how significantly different results could be obtained under different measurement conditions/sample preparation methods, offering optimal protocol for size estimation applying dynamic light scattering (DLS), with complementary analysis through atomic force microscopy. Hydrophilic nanoemulsion and aqueous dispersion of polymeric nanoparticles were used as test samples. Ultrapurified water, phosphate buffer saline (PBS, pH 7.4) and biorelevant medium with serum proteins were used for dilution. Measurements were performed applying batch-mode DLS (ZetasizerNano), and AutoProbe CP-Research microscope. Significant differences in obtained nanodroplet/nanoparticle size were observed depending on the type of medium and dilution level. Protein corona formation could not be confirmed with certainty. Preference was given to PBS as dispersant. The optimal level of dilution for nanoparticles was 1:10 (Z‐ave=59.16±0.46nm), and for nanoemulsion 1:100 (Z‐ave=73.5±0.75nm). For proper interpretation, it is necessary that the DLS measurement report, in addition to the Z‐ave and the polydispersity index, contains at least the data on the attenuation and correlation function intercept. AB - Fizičkohemijske osobine mnogih aktivnih supstanci otežavaju ostvarivanje farmakoloških efekata. Kako bi se identifikovane prepreke prevazišle, visoka očekivanja postavljena su pred nanosisteme - nosače za isporuku aktivnih supstanci, uz značajna ulaganja u njihov razvoj. Nanotehnološki pristup formulaciji leka mnogo je više od pukog smanjenja veličine. Imajući u vidu kompleksnost nanosistema, izazove koji se sreću u njihovoj karakterizaciji, te evidentan nedostatak protokola ispitivanja, relevanta evropska istraživačka/regulatorna tela izdala su vodiče, sumirajući značajne parametre karakterizacije nanosistema. Veličina/distribucija veličina (per se i u biorelevantnim uslovima) od suštinske su važnosti za postizanje očekivanih performansi, predstavljajući kritične atribute kvaliteta (1). Cilj rada bio je pokazati kako se pri različitim uslovima merenja/načinima pripreme istog uzorka dobijaju značajno različiti rezultati procene veličine, te ponuditi optimalan protokol merenja dinamičkim rasipanjem svetlosti (DLS), uz komplementarno ispitivanje mikroskopijom atomskih sila. Kao test uzorci korišćeni su hidrofilna nanoemulzija i vodena disperzija polimernih nanočestica, a kao medijumi za razblaživanje visokoprečišćena voda, izotonični fosfatni pufer (PBS, pH 7,4) i biorelevantni medijum obogaćen proteinima seruma. Merenja su sprovedena primenom batch‐mode DLS, (ZetasizerNano), i na mikroskopu AutoProbe CP-Research. Primećene su značajne razlike u vrednostima veličine nanokapi/nanočestica zavisno od tipa medijuma i nivoa razblaženja. Nije se moglo sa sigurnošću potvrditi formiranje protein korone nakon inkubacije u biorelevantnom medijumu. Za procenu veličine per se, prednost je data PBS-u kao disperzantu. Optimalan nivo razblaženja za nanočestice bio je 1:10 (Z‐ave=59,16±0,46nm), a za nanoemulziju 1:100 (Z‐ave=73,5±0,75nm). Radi adekvatnog tumačenja, potrebno je da izveštaj DLS merenja, pored Z-ave i indeksa polidisperznosti, sadrži barem još podatke o atenuaciji i odsečku korelacione funkcije. PB - Savez farmaceutskih udruženja Srbije (SFUS) C3 - Arhiv za farmaciju T1 - Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles T1 - U susret pravilnom određivanju veličine nanostruktura: analiza uticaja uslova merenja i pripreme uzorka na razultate procene veličine nanokapi i nanočestica VL - 72 IS - 4 suplement SP - S192 EP - S193 UR - https://hdl.handle.net/21.15107/rcub_farfar_4493 ER -
@conference{ author = "Nikolić, Ines and Ranđelović, Danijela and Savić, Snežana", year = "2022", abstract = "Physicochemical properties of many active ingredients jeopardize their pharmacological activity. To overcome identified obstacles, nanosystems as carriers for delivery of actives have been recognized as promising tools, with highly posted expectations. The nanotechnology-based approach in drug formulation is much more than just another step in size miniaturization. Given the complexity of nanosystems, challenges encountered in their characterization, and evident lack of testing protocols, relevant European research/regulatory bodies have issued guidelines, summarizing important parameters for nanosystem characterization. Size/size distribution (per se and in biorelevant environment) are essentially important, representing critical quality attributes (1). The aim was to show how significantly different results could be obtained under different measurement conditions/sample preparation methods, offering optimal protocol for size estimation applying dynamic light scattering (DLS), with complementary analysis through atomic force microscopy. Hydrophilic nanoemulsion and aqueous dispersion of polymeric nanoparticles were used as test samples. Ultrapurified water, phosphate buffer saline (PBS, pH 7.4) and biorelevant medium with serum proteins were used for dilution. Measurements were performed applying batch-mode DLS (ZetasizerNano), and AutoProbe CP-Research microscope. Significant differences in obtained nanodroplet/nanoparticle size were observed depending on the type of medium and dilution level. Protein corona formation could not be confirmed with certainty. Preference was given to PBS as dispersant. The optimal level of dilution for nanoparticles was 1:10 (Z‐ave=59.16±0.46nm), and for nanoemulsion 1:100 (Z‐ave=73.5±0.75nm). For proper interpretation, it is necessary that the DLS measurement report, in addition to the Z‐ave and the polydispersity index, contains at least the data on the attenuation and correlation function intercept., Fizičkohemijske osobine mnogih aktivnih supstanci otežavaju ostvarivanje farmakoloških efekata. Kako bi se identifikovane prepreke prevazišle, visoka očekivanja postavljena su pred nanosisteme - nosače za isporuku aktivnih supstanci, uz značajna ulaganja u njihov razvoj. Nanotehnološki pristup formulaciji leka mnogo je više od pukog smanjenja veličine. Imajući u vidu kompleksnost nanosistema, izazove koji se sreću u njihovoj karakterizaciji, te evidentan nedostatak protokola ispitivanja, relevanta evropska istraživačka/regulatorna tela izdala su vodiče, sumirajući značajne parametre karakterizacije nanosistema. Veličina/distribucija veličina (per se i u biorelevantnim uslovima) od suštinske su važnosti za postizanje očekivanih performansi, predstavljajući kritične atribute kvaliteta (1). Cilj rada bio je pokazati kako se pri različitim uslovima merenja/načinima pripreme istog uzorka dobijaju značajno različiti rezultati procene veličine, te ponuditi optimalan protokol merenja dinamičkim rasipanjem svetlosti (DLS), uz komplementarno ispitivanje mikroskopijom atomskih sila. Kao test uzorci korišćeni su hidrofilna nanoemulzija i vodena disperzija polimernih nanočestica, a kao medijumi za razblaživanje visokoprečišćena voda, izotonični fosfatni pufer (PBS, pH 7,4) i biorelevantni medijum obogaćen proteinima seruma. Merenja su sprovedena primenom batch‐mode DLS, (ZetasizerNano), i na mikroskopu AutoProbe CP-Research. Primećene su značajne razlike u vrednostima veličine nanokapi/nanočestica zavisno od tipa medijuma i nivoa razblaženja. Nije se moglo sa sigurnošću potvrditi formiranje protein korone nakon inkubacije u biorelevantnom medijumu. Za procenu veličine per se, prednost je data PBS-u kao disperzantu. Optimalan nivo razblaženja za nanočestice bio je 1:10 (Z‐ave=59,16±0,46nm), a za nanoemulziju 1:100 (Z‐ave=73,5±0,75nm). Radi adekvatnog tumačenja, potrebno je da izveštaj DLS merenja, pored Z-ave i indeksa polidisperznosti, sadrži barem još podatke o atenuaciji i odsečku korelacione funkcije.", publisher = "Savez farmaceutskih udruženja Srbije (SFUS)", journal = "Arhiv za farmaciju", title = "Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles, U susret pravilnom određivanju veličine nanostruktura: analiza uticaja uslova merenja i pripreme uzorka na razultate procene veličine nanokapi i nanočestica", volume = "72", number = "4 suplement", pages = "S192-S193", url = "https://hdl.handle.net/21.15107/rcub_farfar_4493" }
Nikolić, I., Ranđelović, D.,& Savić, S.. (2022). Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles. in Arhiv za farmaciju Savez farmaceutskih udruženja Srbije (SFUS)., 72(4 suplement), S192-S193. https://hdl.handle.net/21.15107/rcub_farfar_4493
Nikolić I, Ranđelović D, Savić S. Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles. in Arhiv za farmaciju. 2022;72(4 suplement):S192-S193. https://hdl.handle.net/21.15107/rcub_farfar_4493 .
Nikolić, Ines, Ranđelović, Danijela, Savić, Snežana, "Towards Proper Sizing Experiments: Analysis of the Impact of Measurement Conditions and Sample Preparation on the Size Estimation of the Nanodroplets and Nanoparticles" in Arhiv za farmaciju, 72, no. 4 suplement (2022):S192-S193, https://hdl.handle.net/21.15107/rcub_farfar_4493 .