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<script type="text/javascript" src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/jr.boots.min.js"> </script><title>Parameters, Methods and Considerations for the Physicochemical Characterization of Liposomal Products - National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols - NCBI Bookshelf</title>
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<meta name="citation_date" content="2022/06">
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<meta name="citation_author" content="Jeffrey D. Clogston">
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stroke-linecap="round" style="fill:#FFF" d="m320,350a153,153 0 1,0-2,2l170,170m-91-117 110,110-26,26-110-110"></path></svg></a><a id="jr-fip-done" class="wsprkl btn" title="Dismiss find">✘</a></nav><nav id="jr-fip-info-p"><a id="jr-fip-prev" class="wsprkl btn" title="Jump to previuos match">◀</a><button id="jr-fip-matches">no matches yet</button><a id="jr-fip-next" class="wsprkl btn" title="Jump to next match">▶</a></nav></nav></div><div id="jr-epub-interstitial" class="hidden"></div><div id="jr-content"><article data-type="main"><div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><div class="fm-sec"><h1 id="_NBK604275_"><span class="title" itemprop="name">Parameters, Methods and Considerations for the Physicochemical Characterization of Liposomal Products</span></h1><p class="contribs">Clogston JD.</p><p class="fm-aai"><a href="#_NBK604275_pubdet_">Publication Details</a></p></div></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="ncipr75.s1"><h2 id="_ncipr75_s1_">Size/Size Distribution</h2><ul><li class="half_rhythm"><div>Dynamic light scattering (DLS)</div></li><li class="half_rhythm"><div>Multi-angle light scattering (MALS)</div></li><li class="half_rhythm"><div>Laser diffraction</div></li><li class="half_rhythm"><div>Cryogenic-transmission electron microscopy (Cryo-TEM)</div></li><li class="half_rhythm"><div>Resistive pulse sensing</div></li><li class="half_rhythm"><div>Asymmetric-flow field-flow fractionation (AF4) – MALS/DLS</div></li></ul><div id="ncipr75.fig1" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f1&p=BOOKS&id=604275_ncipr75f1.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f1.jpg" alt="Example flow mode DLS spectra with inset of batch mode DLS spectra" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Batch-mode (inset) versus flow-mode DLS measurements of dual-drug loaded liposomes. Multiple size populations are observed by both techniques and indicate a polydispersed sample. However, flow-mode DLS can better resolve the size distribution of each population. Adapted from Anal Bioanal Chem, 2020, 412(2), 425-428.</p></div><div id="ncipr75.s2"><h2 id="_ncipr75_s2_">Composition</h2><ul><li class="half_rhythm"><div>Drug concentration: total, free & encapsulated</div></li><li class="half_rhythm"><div>Drug distribution as a function of size</div></li><li class="half_rhythm"><div>Targeting ligand concentration: total, bound & unbound</div></li><li class="half_rhythm"><div>Individual lipid concentrations</div></li><li class="half_rhythm"><div>Counterion concentrations: interior & exterior</div></li><li class="half_rhythm"><div>Excipient concentrations</div></li><li class="half_rhythm"><div>Particles per mL concentration</div></li><li class="half_rhythm"><div>Osmolality, viscosity measurements</div></li></ul><div id="ncipr75.fig2" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f2&p=BOOKS&id=604275_ncipr75f2.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f2.jpg" alt="Example RP-HPLC chromatogram showing peaks for excipients and ions of PEGylated liposomal doxorubicin" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Counterion and excipient concentrations for PEGylated liposomal doxorubicin measured by RP-HPLC with charged aerosol detection (CAD). Adapted from J Pharm Biomed Anal, 2019, 165, 41-46.</p></div><div id="ncipr75.s3"><h2 id="_ncipr75_s3_">Surface Characteristics</h2><ul><li class="half_rhythm"><div>Zeta potential</div></li><li class="half_rhythm"><div>Protein binding assessment by AF4-MALS/DLS</div></li><li class="half_rhythm"><div>Quartz crystal microbalance with dissipation monitoring (QCM-D)</div></li></ul><div id="ncipr75.fig3" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f3&p=BOOKS&id=604275_ncipr75f3.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f3.jpg" alt="Top graphy shows AF4-MALS spectra and bottom graph shows SF4-DLS spectra" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Flow-mode AF4-MALS (top) and AF4-DLS (bottom) of PEGylated irinotecan liposomes before and after incubation in human plasma. The increase in the ratio (ρ) of the measured MALS (R<sub>g</sub>) and DLS (R<sub>h</sub>) sizes after human plasma incubation suggests protein binding to the surface of the liposomes. Adapted from Anal Bioanal Chem, 2020, 412(2), 425-428.</p></div><div id="ncipr75.s4"><h2 id="_ncipr75_s4_">Purity</h2><ul><li class="half_rhythm"><div>Drug impurities</div></li><li class="half_rhythm"><div>Lipid impurities</div></li><li class="half_rhythm"><div>Free drug/lipid/targeting ligand concentrations</div></li><li class="half_rhythm"><div>Residual solvents and reagents</div></li></ul><div id="ncipr75.fig4" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f4&p=BOOKS&id=604275_ncipr75f4.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f4.jpg" alt="RP-HPLC chromatogram of paclitaxel" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Purity assessment of PEGylated liposomal paclitaxel (PTX) as defined by the presence of drug impurities. The drug epimer concentration was measured by RP-HPLC and its identity confirmed by mass spectrometry.</p></div><div id="ncipr75.s5"><h2 id="_ncipr75_s5_">Morphology</h2><ul><li class="half_rhythm"><div>Cryo-TEM can be used to evaluate:
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<ul><li class="half_rhythm"><div>Size distribution</div></li><li class="half_rhythm"><div>Liposome morphology</div></li><li class="half_rhythm"><div>Internal liposome volume</div></li><li class="half_rhythm"><div>Bilayer thickness</div></li><li class="half_rhythm"><div>Number of lamellae</div></li><li class="half_rhythm"><div>Drug appearance/state</div></li></ul></div></li></ul><div id="ncipr75.fig5" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f5&p=BOOKS&id=604275_ncipr75f5.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f5.jpg" alt="Left shows an electron micrograph of PEGylated liposomal doxorubicin. Right shows two size histograms, top is particle diameter, bottom is bilayer thickness." class="tileshop" title="Click on image to zoom" /></a></div></div><p>Representative cryo-TEM image of PEGylated liposomal doxorubicin. Cryo-TEM was used to determine size and morphology of the liposomes.</p></div><div id="ncipr75.s6"><h2 id="_ncipr75_s6_">Stability</h2><ul><li class="half_rhythm"><div>Size/Size distribution; aggregation</div></li><li class="half_rhythm"><div>Drug leakage and degradation</div></li><li class="half_rhythm"><div>Hydrolysis of lipids</div></li><li class="half_rhythm"><div>Drug release in plasma</div></li><li class="half_rhythm"><div>Solvent, thermal, pH, photo, freeze-thaw, lyophilization, centrifugation, filtration</div></li><li class="half_rhythm"><div>Storage conditions/shelf-life</div></li></ul><div id="ncipr75.fig6" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f6&p=BOOKS&id=604275_ncipr75f6.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f6.jpg" alt="RP-HPLC chromatogram for PEGylated liposomal doxorubicin showing stability of the phopholipids" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Stability assessment of PEGylated liposomal doxorubicin as defined by the hydrolysis of phospholipids. The formation of free fatty acids and lysophospholipids of several batches with varying expiration dates were measured by RP-HPLC with charged aerosol detection (CAD).</p></div><div id="ncipr75.s7"><h2 id="_ncipr75_s7_">Starting Material Characterization</h2><ul><li class="half_rhythm"><div>Drug identity (structure)</div></li><li class="half_rhythm"><div>Drug purity (degradation products)</div></li><li class="half_rhythm"><div>Lipid composition (structure, fatty acid distribution)</div></li><li class="half_rhythm"><div>Lipid purity (free fatty acid, lysophospholipids)</div></li><li class="half_rhythm"><div>Storage conditions/shelf life</div></li></ul><div id="ncipr75.fig7" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f7&p=BOOKS&id=604275_ncipr75f7.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f7.jpg" alt="RP-HPLC chromatogram of common commercial lipid mix for liposome samples" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Lipid composition (identity and individual lipid concentrations) and purity (presence of free fatty acid and lysophospholipids) were determined by RP-HPLC with charged aerosol detection (CAD) for a commercially available lipid mix. The theoretical mass ratio was confirmed.</p></div><div id="ncipr75.s8"><h2 id="_ncipr75_s8_">Batch-to-Batch Consistency</h2><ul><li class="half_rhythm"><div>Assessed by choosing relevant parameters (i.e., lot release criteria) that relate to a desired in vivo outcome</div></li></ul><div id="ncipr75.fig8" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f8&p=BOOKS&id=604275_ncipr75f8.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK604275/bin/ncipr75f8.jpg" alt="RP-HPLC chromatogram of a lipid nanoparticle" class="tileshop" title="Click on image to zoom" /></a></div></div><p>Batch-to-batch consistency for lipid nanoparticles with siRNA was assessed by quantitation of the lipid composition. Six individual lipid concentrations for three batches were determined by RP-HPLC with charged aerosol detection (CAD).</p></div><div id="ncipr75.rl.r1"><h2 id="_ncipr75_rl_r1_">Relevant NCL Publications</h2><ol><li><div class="bk_ref" id="ncipr75.ref1"><em>Anal Bioanal Chem</em>, 2020, 412(2), 425–428. PMID: 31776639
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[<a href="https://pubmed.ncbi.nlm.nih.gov/31776639" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 31776639</span></a>]</div></li><li><div class="bk_ref" id="ncipr75.ref2"><em>J Control Release</em>, 2019, 299, 31–43. PMID: 30797868
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[<a href="https://pubmed.ncbi.nlm.nih.gov/30797868" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 30797868</span></a>]</div></li><li><div class="bk_ref" id="ncipr75.ref3"><em>J Pharm Biomed Anal</em>, 2019, 165, 41–46. PMID: 30502551
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[<a href="/pmc/articles/PMC6331219/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC6331219</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/30502551" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 30502551</span></a>]</div></li><li><div class="bk_ref" id="ncipr75.ref4"><em>Pharmaceutical Research</em>, 2019, 37, 6. PMID: 31828540
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[<a href="/pmc/articles/PMC7274461/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC7274461</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/31828540" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 31828540</span></a>]</div></li><li><div class="bk_ref" id="ncipr75.ref5"><em>Methods in Molecular Biology</em>, Vol. 1628, 2018, p. 49–55. PMID: 29039092
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[<a href="https://pubmed.ncbi.nlm.nih.gov/29039092" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 29039092</span></a>]</div></li><li><div class="bk_ref" id="ncipr75.ref6"><em>Anal Bioanal Chem</em>, 2017, 409(24), 5779–5787. PMID: 28762066
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[<a href="https://pubmed.ncbi.nlm.nih.gov/28762066" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 28762066</span></a>]</div></li></ol></div><div><dl class="temp-labeled-list small"><dl class="bkr_refwrap"><dt></dt><dd><div><p class="no_top_margin">
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<b>About NCL</b>
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</p><p>The Nanotechnology Characterization Laboratory (NCL) is a resource for nanotech researchers and organizations developing nano-based therapies and diagnostics. The NCL provides preclinical characterization services through various collaboration mechanisms. Learn more by visiting our website: <a href="https://ncl.cancer.gov" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">https://ncl<wbr style="display:inline-block"></wbr>​.cancer.gov</a></p><p><b>Email us at</b>
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<a href="mailto:dev@null" data-email="vog.hin.liam@lcn" class="oemail">vog.hin.liam@lcn</a></p></div></dd></dl></dl></div><div id="bk_toc_contnr"></div></div></div><div class="fm-sec"><h2 id="_NBK604275_pubdet_">Publication Details</h2><h3>Author Information and Affiliations</h3><p class="contrib-group"><h4>Authors</h4><span itemprop="author">Jeffrey D. Clogston</span>.</p><h3>Publication History</h3><p class="small">Published: <span itemprop="datePublished">June 2022</span>.</p><h3>Copyright</h3><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div></div><h3>Publisher</h3><p><a href="https://www.cancer.gov/nano/research/ncl" ref="pagearea=page-banner&targetsite=external&targetcat=link&targettype=publisher">National Cancer Institute (US)</a>, Bethesda (MD)</p><h3>NLM Citation</h3><p>Clogston JD. Parameters, Methods and Considerations for the Physicochemical Characterization of Liposomal Products. 2022 Jun. In: National Cancer Institute’s Nanotechnology Characterization Laboratory Assay Cascade Protocols [Internet]. Bethesda (MD): National Cancer Institute (US); 2005 May 1-. <span class="bk_cite_avail"></span> doi: 10.17917/C7201Q</p></div><div class="small-screen-prev"><a href="/books/n/nciprotocols/ncipr76/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M75,30 c-80,60 -80,0 0,60 c-30,-60 -30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Prev</text></svg></a></div><div class="small-screen-next"><a href="/books/n/nciprotocols/ncipr74/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M25,30c80,60 80,0 0,60 c30,-60 30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Next</text></svg></a></div></article><article data-type="fig" id="figobncipr75fig1"><div id="ncipr75.fig1" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f1&p=BOOKS&id=604275_ncipr75f1.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f1.jpg" alt="Example flow mode DLS spectra with inset of batch mode DLS spectra" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig2"><div id="ncipr75.fig2" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f2&p=BOOKS&id=604275_ncipr75f2.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f2.jpg" alt="Example RP-HPLC chromatogram showing peaks for excipients and ions of PEGylated liposomal doxorubicin" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig3"><div id="ncipr75.fig3" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f3&p=BOOKS&id=604275_ncipr75f3.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f3.jpg" alt="Top graphy shows AF4-MALS spectra and bottom graph shows SF4-DLS spectra" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig4"><div id="ncipr75.fig4" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f4&p=BOOKS&id=604275_ncipr75f4.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f4.jpg" alt="RP-HPLC chromatogram of paclitaxel" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig5"><div id="ncipr75.fig5" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f5&p=BOOKS&id=604275_ncipr75f5.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f5.jpg" alt="Left shows an electron micrograph of PEGylated liposomal doxorubicin. Right shows two size histograms, top is particle diameter, bottom is bilayer thickness." class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig6"><div id="ncipr75.fig6" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f6&p=BOOKS&id=604275_ncipr75f6.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f6.jpg" alt="RP-HPLC chromatogram for PEGylated liposomal doxorubicin showing stability of the phopholipids" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig7"><div id="ncipr75.fig7" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f7&p=BOOKS&id=604275_ncipr75f7.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f7.jpg" alt="RP-HPLC chromatogram of common commercial lipid mix for liposome samples" class="tileshop" title="Click on image to zoom" /></a></div></div></article><article data-type="fig" id="figobncipr75fig8"><div id="ncipr75.fig8" class="figure"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Image%20ncipr75f8&p=BOOKS&id=604275_ncipr75f8.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK604275/bin/ncipr75f8.jpg" alt="RP-HPLC chromatogram of a lipid nanoparticle" class="tileshop" title="Click on image to zoom" /></a></div></div></article></div><div id="jr-scripts"><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/libs.min.js"> </script><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/jr.min.js"> </script></div></div>
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