Fasted-State Simulated Intestinal Fluid "FaSSIF-C", a Cholesterol Containing Intestinal Model Medium for In Vitro Drug Delivery Development Pooneh Khoshakhlagh,

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Fasted-State Simulated Intestinal Fluid "FaSSIF-C", a Cholesterol Containing Intestinal Model Medium for In Vitro Drug Delivery Development Pooneh Khoshakhlagh, Raphael Johnson, Peter Langguth, Thomas Nawroth, Lars Schmueser, Nadja Hellmann, Heinz Decker, Noemi Kinga Szekely Journal of Pharmaceutical Sciences Volume 104, Issue 7, Pages 2213-2224 (July 2015) DOI: 10.1002/jps.24470 Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 1 Solubility of (a) Fenofibrate, (b) Carbamazepine, (c) Danazol, and (d) Griseofulvin in gastrointestinal model fluids FaSSIF-C containing various amounts of cholesterol. The physiologically related fluids contain 7% (female), 10% (male), and 13% (disease) cholesterol (mol % of amphiphiles; sodium TC + lecithin+ cholesterol); the range for healthy persons is indicated by the box. The difficult samples of high cholesterol content (13%) are qualitatively shown for comparison only. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 2 Average nanoparticle size of gastrointestinal model fluids FaSSIF-C containing various amounts of cholesterol without drug before (▲) and after centrifugation (■). At high cholesterol content, additional large particles appear as minor components, which are indicated for the samples after centrifugation as symbols off size scale (+, *, o). The difficult samples of high cholesterol content (13%) are qualitatively shown for comparison only. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 3 Detailed structure analysis (one individual sample experiment) of nanoparticles in gastrointestinal model fluid FaSSIFmod6.5 by SANS: (a) the SANS scattering profile appears as a sum of several particle contributions; the size analysis by Guinier plots of (b) 8m data (3-h-old sample), and (c) 2m data (47-min-old sample) indicate particles of different size in parallel. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 4 Particle size profiles of Fenofibrate in FaSSIF-10C (10% cholesterol) obtained by DLS in (a) linear and (b) logarithmic representation of the volume contribution (one individual sample experiment). The minor contributions are visible only in the unusual logarithmic plot. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 5 Average nanoparticle size (three sample average) of gastrointestinal model fluids FaSSIF-C containing various amounts of cholesterol and (a) Fenofibrate, (b) Carbamazepine, (c) Danazol, and (d) Griseofulvin. At high cholesterol content, additional large particles appear as minor components, which are indicated as symbols off size scale (+, *, o). Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 6 The biorelevant model fluid FaSSIF-7C and conventional FaSSIFmod6.5 intestinal model fluids do not damage gut cells (Caco-2) upon incubation for 1h, whereas the bare transfer medium TM reduces the viability by 20%. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions

Figure 7 The nanoparticles in the simulated intestinal segment develop from micelles to liposomes, and some large particles in the presence of cholesterol. The drug solubility changes in partly parallel, which is an evidence for nanointermediates. Journal of Pharmaceutical Sciences 2015 104, 2213-2224DOI: (10.1002/jps.24470) Copyright © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association Terms and Conditions