Measuring Primary and Secondary Nucleation in Crystallization

Joop H. ter HorstEPSRC Centre for Innovative Manufacturing in Continuous Manufacturing and Crystallisation (CMAC)Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS)Technology and Innovation CentreUniversity of Strathclyde99 George Street, Glasgow G1 1RD, U.K.

Email: Joop.terHorst@strath.ac.uk

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Ter Horst Group

Technology & Innovation Centre

Prof. Joop H. ter HorstEPSRC Future Manufacturing Research Hub (CMAC)Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS)Technology and Innovation CentreUniversity of Strathclyde99 George Street, Glasgow G1 1RD, U.K.

Phone: +44 141 548 2858Email: Joop.terHorst@strath.ac.ukwww.strath.ac.ukwww.cmac.ac.ukwww.coreitn.eu

Strathclyde Institute of Pharmacy and Biomedical Sciences

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Ter Horst Group

Industrial Crystallization• Continuous Crystallization

– Andrew Dunn, Sebastion Davidson

• Crystallization Kinetics in Complex Multicomponent Systems– Olayinka Olalere, Corin Mack, Suse Bebiano, Carlos Moreno, Dr.

Stephanie Urwin

• Protein Crystallization Technology– Dr. Maria Briuglia, Dr. Charline Gerard

• Chiral Resolution– Johannes Hoffmann, Raghu Venkatramanan, Giulio Valenti, Jose Luis

Capdevila Echeverria, Weiwei Li (Delft University of Technology)

Crystal StructureSize

ShapePurity

Crystallization is a highly efficient Separation Technology

With the added advantage that it results in a Particulate Product

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Solution Primarynucleation crystal growth

Secondary nucleation

agglomeration

supersaturation

hydrodynamics

solid formcrystal size

crystal shapepurity

Product quality

Industrial Crystallization

Productivity

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Crystallization Design WorkflowC.J. Brown, J.H. ter Horst, …, Enabling Precision Manufacturing of Active Pharmaceutical Ingredients: Workflow for Seeded Cooling Continuous Crystallisations. Molecular Systems Design Engineering (2018)

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Crystallization Design WorkflowC.J. Brown, J.H. ter Horst, …, Enabling Precision Manufacturing of Active Pharmaceutical Ingredients: Workflow for Seeded Cooling Continuous Crystallisations. Molecular Systems Design Engineering (2018)

The first hurdle towards Crystallization Process Design:

Know the solubility

Measuring Primary and Secondary Nucleation in Crystallization• Solubility• Primary Nucleation• Secondary Nucleation

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Crystallization from SolutionEvaporative Crystallization Cooling Crystallization Antisolvent Crystallization

Precipitation

J.H. ter Horst, C. Schmidt, J. Ulrich, Fundamentals of Industrial Crystallization, In: Nishinaga T, Rudolph P, editors, Handbook of Crystal Growth, Vol. II., Elsevier, 2015, pp. 1317–49, DOI: 10.1016/B978-0-444-63303-3.00032-8.

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Crystallization from Solution

C = 100 mg/ml solventC* = 20 mg/ml solvent

Yield C-C* = 80 mg crystals/ml solvent

80% is crystallized,20% remains in solution

Concentration CSolubility C*

[mg/ml]

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Crystal Solubility Measurement

100% pureCrystalline phase

Crystal solubility C*:The solution concentration that is in equilibrium with the crystalline solid at a

specific temperature T and pressure P.

Solution withConcentration C*At temperature T,

Pressure T

EquilibriumSolubility behaviour determines crystallization method & Yield

How to measure solubility?

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Temperature Variation Method

• Increase solubility until suspension turns into a clear solution• Reproducing results fairly quick• Also metastable zone width

Temperature [°C]

Change solubility

Concentration[mg/mL]

Clear point temperature

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Clear & Cloud Point Measurements

T

Transmission

Clear point, 100% transmission

Ts=42.2 CTs=42.3 C

1440 min = 1 dayHeating rate = 0.3°C/min

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Clear Point & Solubility

ThermodynamicSaturation temperature

Concentration = Constant

If:• Crystal detection limit is low• Dissolution is fast• No fouling• No crowning• …

check the vialsDuring the

measurement!Often, a heating rate of

0.3°C/min gives sufficiently accurate data

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Solubility DiagramOf isonicotinamide (INA) in Ethanol

J.H. ter Horst, M.A. Deij, P.W. Cains, Discovering new co-crystals,

Crystal Growth Design 9(3) (2009) 1531-1537.

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Supersaturation RatioOf isonicotinamide (INA) in Ethanol

12.5°C

Solubility x*

Concentration x

S = xx*

Measuring Primary and Secondary Nucleation in Crystallization• Solubility• Primary Nucleation• Secondary Nucleation

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Primary Nucleation Characteristics

Develop a method for measuring Primary Nucleation Rate

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Primary Nucleation Characteristics

Clear point - Upon heating there is a temperature that a suspension turns into a clear solutionCloud point - Upon cooling a solution there is a temperature that crystals will be detectedMetastable Zone Width - The difference between the saturation temperature (Clear point) and cloud point

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Metastable zone width

0

25

50

75

100

0

10

20

30

40

50

60

6:00 7:00 8:00 9:00 10:00 11:00 12:00

Time [hrs]

Temperature[ C]

Transmissionof light

[%]MSZW

Clear PointTemperature

Cloud PointTemperature

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Measuring Induction TimesThe time until detection of crystals at constant supersaturation

• Accurate temperature control = accurate supersaturation control• A certain minimum volume fraction of crystals in the stirred suspension is

needed for detection

0

25

50

75

100

20

30

40

50

60

7 8 9 10 11 12

Transmission [%]

Temperature [oC]

Time [hr]

t

1 mlStirred solutions

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Measuring Induction TimesS. Jiang, J.H. ter Horst, Crystal Growth Design 11 (2011) 256-261

Constant S • 1ml stirred solutions

• Create constant supersaturation

• Measure induction time

• Do this a large number of times

Crystal nucleation rate is very low

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Single Nucleus Mechanism

Observed in 3 ml stirred solutions for:• Isonicotinamide in Ethanol, butanol, nitrobenzene, nitromethane• Paracetamol, succinic acid in water• Hydroxyacetophenone in acetyl acetate

S.S. Kadam, J.H. ter Horst et al., Cryst. Growth Des. 11(4) (2011) 1271–1277

Parent crystal Crystal suspension

Clear solution

Secondarynucleation

PrimaryHeterogeneous

nucleationGrowth

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Measuring Induction TimesS. Jiang, J.H. ter Horst, Crystal Growth Design 11 (2011) 256-261

P(t)=M +(ti)/MConstant S

P(t)= 1 - exp(-JV(t - tg))

J = 630±20 m-3s-1

tg = 1170±20 s

Model:

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Induction Time Distributions

m-ABA in 50 wt % water/ethanol L-His in water Isonicotinamide in ethanol

S. Jiang, J.H. ter Horst, Crystal Growth Design 11 (2011) 256-261S. Kulkarni, J.H. ter Horst, Crystal Growth Design 13 (2013) 2435-2440

S=1.83

S

S=2.15

S=1.55

S

S=1.74

S=1.264

S

S=1.48

Supersaturation S = Concentration / Solubility

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Induction Time Distributions

Nucleation of Metal Organic Framework

0

0.25

0.5

0.75

1

0 40000 80000 120000

P(t)

t [s]

With Silica ParticlesWithout Silica

Nucleation of Proteins

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Nucleation Rates

In IPA In DMF

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Crystal Nucleation Rates Analysis

System A [m-3s-1] BDPL form RII in IPA 576 0.68DPL form RI in DMF 499 4.57

• Nucleation work B/ln2S in DMF much higher

• Pre-exponential factors A very low => heterogeneous nucleation

2explnBJ ASS

æ ö= -ç ÷è ø

LowSHighS

J.H. ter Horst, C. Brandel, Measuring Induction Times and Crystal Nucleation Rates, Faraday Discuss. 179 (2015) 199

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!

□ - Untreated sample○ - Solution in silanized vials∆ - Filtered solution

Filtration removes (part of) the heterogeneous particles

Nucleation occurs in bulk of solutiononto heterogeneous (dust) particle

Heterogeneous Crystal Nucleation

Crystal Nucleation Rate Mechanism

P(t)

t [s]

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Heterogeneous Nucleationof CO2 bubbles on a Mentos in Diet Coke

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Crystal Nucleation

nucleation

primary secondary

homogeneous heterogeneous

Measuring Primary and Secondary Nucleation in Crystallization• Solubility• Primary Nucleation• Secondary Nucleation

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Secondary Nucleation is the formation of particles due to the presence of other particles

Attrition Fluid Shear

Develop a systematic method for secondary nucleation assessment

Secondary Nucleation

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Metastable Zone WidthOf isonicotinamide (INA) in Ethanol

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Induction time measurementsOf isonicotinamide (INA) in Ethanol

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Induction time measurementsOf isonicotinamide (INA) in Ethanol

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Single Crystal Seed Experiment

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Calibrating Suspension Density

0

50

100

150

200

0 50 100 150

N[#]

Nρx 103 [#/ml]

Number N of particles determined by Crystalline software versus suspension density N!

50 μm monodisperse polymer particles

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Seeded experiment Unseeded experiment

Single Crystal Seed Experiment

N!x1000[#/ml]

Suspension density in time after seeding single crystal

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Seeded experiment Unseeded experiment

Single Crystal Seed Experiment

N!x1000[#/ml]

Suspension density in time after seeding single crystal

Secondary Nucleation Rate B=

Slope of suspension density in time

B = 12.8 . 103 #/ml.min

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Seeds size: 1.7 – 2.4 mm

Secondary nucleation rate

Bx1000

[#/ml.min]

Secondary Nucleation threshold

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Crystallization Design Workflow

C.J. Brown, …, J.H. ter Horst, …, Enabling Precision Manufacturing of Active Pharmaceutical Ingredients: Workflow for Seeded Cooling Continuous Crystallisations. Molecular Systems Design Engineering (2018) https://doi.org/10.1039/C7ME00096K.

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Secondary Nucleation Understanding

Metastable Zone Width

Choose Supersaturation S

Measure Induction Times at Selected S

Single Crystal Seeds Preparation

Measure Secondary Nucleation Rates at Selected S

Secondary Nucleation Threshold

Measuring Primary and Secondary Nucleation in Crystallization• Solubility• Primary Nucleation• Secondary Nucleation

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Acknowledgements

Solubility• Marloes Reus

Primary Nucleation• Shanfeng Jiang, Samir

Kulkarni, Maria Briuglia, Clement Brandel

Secondary Nucleation• Maria Briuglia, Jan Sefcik

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LiteratureSolubility• J.H. ter Horst, M.A. Deij, P.W. Cains, Discovering new co-crystals, Crystal Growth Design 9(3) (2009) 1531-1537. • S. Srisanga, J.H. ter Horst, Racemic Compound, Conglomerate, or Solid Solution: Phase Diagram Screening of Chiral Compounds, Crystal Growth

Design 10(4) (2010) 1808-1812.• J. Vellema, N.G.M. Hunfeld, H.E.A. Van den Akker, J.H. ter Horst, Avoiding Crystallization of Lorazepam during Infusion, Eur. J. Pharm. Sci. 44 (2011)

621–626.• António O.L. Évora, Ricardo A.E. Castro, Teresa M.R. Maria, M. Ramos Silva, J.H. ter Horst, João Canotilho, M. Ermelinda S. Eusébio, Thermodynamic

based approach on the investigation of a diflunisal pharmaceutical co-crystal with improved intrinsic dissolution rate, International Journal of Pharmaceutics 466 (2014) 68–75.

• M.A. Reus, A.E.D.M. van der Heijden, J.H. ter Horst, Solubility Determination from Clear Points upon Solvent Addition, Org. Process Res. Dev. 19 (8) (2015) 1004–1011, DOI: 10.1021/acs.oprd.5b00156

• A.O.L. Evora, R. Castro, T. Maria, M.R. Silva, J.H. ter Horst, J. Canotilho, M.E.S. Eusébio, Co-crystals of diflunisal and isomeric pyridinecarboxamides–a thermodynamics and crystal engineering contribution, CrystEngComm, 18 (2016) 4749–4759, DOI: 10.1039/c6ce00380j (May 3, 2016).

Primary Nucleation• S. Jiang, J.H. ter Horst, Crystal Nucleation Rates from Probability Distributions of Induction Times, Crystal Growth Design 11 (2011) 256-261.• S.S. Kadam, S.A. Kulkarni, R. Coloma Ribera, A.I. Stankiewicz, J.H. ter Horst, Herman J.M. Kramer, A new view on the metastable zone width during

cooling crystallization, Chem. Eng. Sci. 72 (2012) 10–19.• S.A. Kulkarni, S.S. Kadam, H. Meekes, A.I. Stankiewicz, J.H. ter Horst, Crystal Nucleation Kinetics from Induction Times and Metastable Zone Widths,

Crystal Growth Design 13(6) (2013) 2435-2440.• R.J. Davey, S.L.M. Schroeder, J.H. ter Horst, Nucleation of Organic Crystals – A Molecular Perspective, Angewandte Chemie International Edition 52

(2013) 2166-2179, DOI: 10.1002/anie.201204824.• R. A. Sullivan, R. J. Davey, G. Sadiq, G. Dent, K. R. Back, J. H. ter Horst, D. Toroz, R. B. Hammond, Revealing the Roles of Desolvation and Molecular

Self-Assembly in Crystal Nucleation from Solution: Benzoic and p-Aminobenzoic Acid, Cryst. Growth Des. 14 (2014) 2689−2696.• Antonella Caridi, Samir A. Kulkarni, Gianluca Di Profio, Efrem Curcio, Joop H. ter Horst, Template-Induced Nucleation of Isonicotinamide

Polymorphs, Cryst. Growth Des. 14 (2014) 1135−1141.• J.H. ter Horst, C. Brandel, Measuring Induction Times and Crystal Nucleation Rates, Faraday Discuss. 179 (2015) 199• C. Brandel, Y. Cartigny, S. Petit, J.H. ter Horst, G. Coquerel, Pre-Nucleation Self-Assembly and Chiral Discrimination Mechanisms during Solution

Crystallization of Racemic Diprophylline, Chemistry-A European Journal 22 (2016) 16103-16112, DOI: 10.1002/chem.201602707 (September 26, 2016).

• H. Yang, J.H. ter Horst, Crystal Nucleation of Small Organic Molecules, pp 317-337, In: New Perspectives on Mineral Nucleation and Growth, Editors: A.E.S. Van Driessche, M. Kellermeier, L.G. Benning, Denis Gebauer (2017), Springer International Publishing, DOI: 10.1007/978-3-319-45669-0_16.

Secondary Nucleation• C.J. Brown, …, J.H. ter Horst, …, Enabling Precision Manufacturing of Active Pharmaceutical Ingredients: Workflow for Seeded Cooling Continuous

Crystallisations. Molecular Systems Design Engineering (2018) https://doi.org/10.1039/C7ME00096K.• M. Briuglia, J. Sefcik, J.H. ter Horst, submitted

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