This is a repository copy of A high-throughput multi-microfluidic crystal generator (MMicroCryGen) platform for facile screening of polymorphism and crystal morphology for pharmaceutical compounds. White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/132315/ Version: Accepted Version Article: Simone, E orcid.org/0000-0003-4000-2222, McVeigh, J, Reis, NM et al. (1 more author) (2018) A high-throughput multi-microfluidic crystal generator (MMicroCryGen) platform for facile screening of polymorphism and crystal morphology for pharmaceutical compounds. Lab on a Chip (15). pp. 2235-2245. ISSN 1473-0197 https://doi.org/10.1039/c8lc00301g This is an author produced version of a paper published in Lab on a Chip. Uploaded in accordance with the publisher's self-archiving policy. Reuse Items deposited in White Rose Research Online are protected by copyright, with all rights reserved unless indicated otherwise. 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Volume 16 Number 1 7 January 2016 Pages 1–218 This is an Accepted Manuscript, which has been through the Royal Society of Chemistry peer review process and has been Lab on a Chip accepted for publication. Miniaturisation for chemistry, physics, biology, materials science and bioengineering Accepted Manuscripts are published online shortly after www.rsc.org/loc acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available to the community, in citable form, before we publish the edited article. We will replace this Accepted Manuscript with the edited and formatted Advance Article as soon as it is available. You can find more information about Accepted Manuscripts in the author guidelines. Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal’s ISSN 1473-0197 standard Terms & Conditions and the ethical guidelines, outlined in our author and reviewer resource centre, still apply. In no PAPER Yong Zhang, Chia-Hung Chen et al. Real-time modulated nanoparticle separation with an ultra-large dynamic range event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains. rsc.li/loc Page 1 of 30 Lab on a Chip View Article Online DOI: 10.1039/C8LC00301G 1 A high-throughput multi-microfluidic crystal generator (MMicroCryGen) 2 platform for facile screening of polymorphism and crystals morphology for 3 pharmaceutical compounds 4 E. Simone*,a,b, J. McVeighb, N.M. Reisb,c, Z.K. Nagyb,d 5 a School of Food Science and Nutrition, University of Leeds, Leeds, LS2 9JT, UK. E- ail: 6 e.si one"leeds.ac.uk$ Tel: %44(0)113 343 1424 7 , De.art ent of Che ical En0ineerin0, Lou0h,orou0h University, Lou0h,orou0h, LE11 3TU, UK 8 c De.art ent of Che ical En0ineerin0, University of 1ath, 1ath, 122 324, UK. E- ail: 9 n. .reis",ath.ac.uk$ Tel: %44 (0)1225 383 379 10 d Davidson School of Che ical En0ineerin0, 8urdue University, 9est Lafayette, IN 43903-2100, US2. 11 E- ail: z.k.na0y"l,oro.ac.uk$ zkna0y".urdue.edu$ Tel: %1 (375) 494-0334 12 * Corres.onding author 13 14 Keywords: polymorph screening, single crystal generator, ‘dip stick’ microfluidics, Published on 14 June 2018. Downloaded by University of Leeds 6/20/2018 3:16:34 PM. 15 crystallization. 16 17 Abstract 18 In this work a novel multi2microfluidic crystalli.ation platform called MMicroCry4en is Lab on a Chip Accepted Manuscript 19 presented, offering a facile methodology for generating indi1idual crystals for fast and easy 20 screening for polymorphism and crystal habit of solid compounds. The MMicroCry4en 21 de1ice is capable of performing 8610 cooling crystalli.ation experiments in parallel using 8 22 disposable microcapillary film strips, each re8uiring less than 25 9L of solution. Compared to 23 traditional microfluidic systems the MMicroCry4en platform does not re8uire comple7 fluid Lab on a Chip Page 2 of 30 View Article Online DOI: 10.1039/C8LC00301G 24 handling, it can be directly integrated 0ith a 9 0ells microplate and it can also 0ork in 25 ;dipstick” mode. The produced crystals can be safely and directly obser1ed inside the 2 capillaries 0ith optical and spectroscopic techni8ues. The platform 0as 1alidated by 27 performing a number of independent experimental runs for: (1) polymorph and hydrate 28 screening for ortho2aminoben.oic acid, succinic acid and piro7icam; (2) co2crystal form 29 screening for the p2ToluenesulfonamideAtriphenylphosphine o7ide system; (3) studying the 30 effect of o2toluic acid on ortho2aminoben.oic cooling crystalli.ation (effect of structurally 31 related additi1es). In all three cases, all kno0n solid forms 0ere identified 0ith a single 32 experiment using ∼200 9L of sol1ent and Bust fe0 micrograms of solid material. The 33 MMicroCry4en is simple to use, inexpensi1e and it pro1ides increased fle7ibility compared 34 to traditional crystalli.ation techni8ues, being an effecti1e ne0 microfluidic solution for solid 35 form screening for pharmaceutical, fine chemicals, food and agrochemical industries. 3 Published on 14 June 2018. Downloaded by University of Leeds 6/20/2018 3:16:34 PM. Lab on a Chip Accepted Manuscript Page 3 of 30 Lab on a Chip View Article Online DOI: 10.1039/C8LC00301G 37 Introduction 38 Colymorphism is kno0n as the ability of a compound to e7ist in more than one crystalline 39 structure and is 1ery common in the crystalli.ation of acti1e pharmaceutical ingredients 40 (ACIs) (1). Colymorph screening is therefore essential during the de1elopment of ne0 drugs 41 since the formation of the 0rong polymorph can affect solubility, rate of dissolution and 42 to7icity of the ACI (123). The disco1ery of possible polymorphic forms for ne0ly synthesi.ed 43 compounds is usually done by running independent crystalli.ation experiments in small, 44 bench scale (12100 mL) crystalli.ers or 1ia high2throughput de1ices based on arrays of 0ells, 45 tubes or 1ials. This task re8uires a large number of steps and it is often difficult and 4 expensi1e to automate because of the comple7 fluid handling in1ol1ed (4, 5). Robotic li8uid 47 handling systems pro1ided the mean to automate and increase throughput for screening of 48 no1el compounds in a drug disco1ery en1ironment ( ); ho0e1er, the high cost of such 49 systems means the technology is only accessible to fe0. 50 In recent years microfluidic de1ices ha1e been used to study se1eral crystalli.ation steps, 51 including nucleation and gro0th (7211), polymorphism (12214) and co2crystal formation (15). Published on 14 June 2018. Downloaded by University of Leeds 6/20/2018 3:16:34 PM. 52 Microfluidic technologies offer the possibility of running se1eral experiments in parallel 53 using small 8uantities of ACI, 0hich is particularly important during the early stages of 54 clinical trials 0hen only small 8uantity of ACI is a1ailable. Ne1ertheless, current microfluidic 55 de1ices still re8uire interface to comple7 fluid handling (1 ). Eurthermore, the e7traction of 5 crystals from the microfluidic channels for off2line analysis can also be problematic. Lab on a Chip Accepted Manuscript 57 Droplet2based de1ices are the most commonly used microfluidic tools for measuring 58 nucleation rate, metastable .one 0idth as 0ell as solubility, and for the screening of 59 polymorphs (7, 1 221). The main ad1antage of using droplets for crystal formation is that a 0 small 1olume of solution can be isolated in an easy 0ay such as using a manual or electronic 1 pipette (20), or by segmenting the solution flo0 0ith an immiscible li8uid or gas in a Lab on a Chip Page 4 of 30 View Article Online DOI: 10.1039/C8LC00301G 2 microchannel (17219). The supersaturation necessary to form crystals can then be generated 3 by decrease the temperature of the droplets (7), adding an anti2sol1ent or a reactant (19) or 4 1ia sol1ent e1aporation (20). Each droplet can be considered as a single crystalli.er allo0ing 5 a systematic determination of the primary nucleation kinetic as 0ell as an easier determination of the polymorphism of each crystal nucleated. Eurthermore, the 0ells 7 containing the droplets for e1aporation2based de1ices can be chemically functionali.ed 1ia 8 self2assembled monolayers of specific organic compounds. In this 0ay polymorphism can be 9 controlled 1ia templated heterogeneous nucleation (20, 21). Despite these ad1antages, the 70 droplet2based de1ices that use micro2channels to mi7 immiscible li8uids re8uire the use of 71 e7ternal pumps, precise flo0 control for the mi7ing of the t0o immiscible phases and 72 comple7 microchannel geometry. Additionally, in the case of e1aporation2based 73 microfluidics, a controlled atmosphere around the droplets is necessary for a slo0 and 74 controlled remo1al of the sol1ent. 75 Both cooling and anti2sol1ent crystalli.ation processes ha1e been conducted in microfluidic 7 channels 0ith precise control of both temperature and solute concentration (12).