Antisolvent Crystallisations

Gerry Steele
 Antisolvent Crystallisations

• First alternative to cooling crystallisation when

it does not work due to:
• Solubility
• Not enough temperature dependence
• Very high or very low solubility in most
solvents
• Can be used to increase yield
70

60 IPrOAc:Heptane (100:0)
IPrOAc:Heptane (80:20)
IPrOAc:Heptane (50:50)
50

Solubility (mg/mL)
40

30

20

10

0
0 5 10 15 20 25 30 35 40 45 50
Temperature (°C)

• Isopropyl acetate was the reaction solvent

and the free base was crystallised from that
solution.
• Changing from pure IPrOAc to a 50:50
mixture with heptane, increased the yield
from 75 % to about 92 %.
 Development of an Anti-Solvent Addition
Crystallisation

• With knowledge of the solubility data with respect to solvent

composition the following can be determined and evaluated
– an acceptable final solvent composition
– the impact on the yield of the process with respect to solvent volumes
– determination of the outline process with respect to the desired yield
and process volumes
– minimise the amount of anti-solvent that is added during crystallisation
– the amount of anti-solvent that can be added in one portion with no
effect on the control of the process
– the portion of anti-solvent that needs to be added in a controlled manner
Solvent Selection
Prat et al. Green Chem 16 4546-4551 (2014)
 Solvent Selection-ICH Guidelines

• International Committee on Harmonization has

issued guidelines (Q3c) on solvent usage
• Three classes defined
• Class 1: Solvents to be avoided, e.g. benzene,
carbon tetrachloride
• Class 2: Solvents to be limited, e.g.
cyclohexane, toluene, acetonitrile
• Class 3: solvents with low toxic potential,
acetone, ethanol, ethyl acetate
• Others for which no adequate toxicological data
on which a PDE has been found. Need to
supply a justification for its use, e.g. Di-iso-
propyl ether
 Typical Solvent/Anti-Solvent Systems
• Polar
– Methanol/water, ethanol/water, IPA/water, methanol/IPA
• Mixed
– methanol/toluene, ethanol/toluene, ethanol/ethyl acetate,
ethanol/iso-propyl acetate, acetone/acetonitrile,
acetone/ethyl acetate, IPA/ethyl acetate
• Non-polar
– IPA,MEK, ethyl acetate, iso-propyl acetate, toluene, butyl
acetate and
– heptane or iso-octane or cyclohexane
• Check miscibility !
 Solvent Miscibility

• Not all solvents are

miscible with each
other, but some solvents
will dissolve in each
other to a certain
degree
• For example, 3.3 mL of
water will dissolve in 90
mL of anhydrous ethyl
acetate (and cause a
change of volume)
• Indeed, anhydrous ethyl
acetate adsorbs water
readily from the
atmosphere
 Solvent-Water Mixes

• The presence of water in solvents

can have implications in a batch
process, i.e. there may be batch to
batch variation in the water content
of solvents
• The water activity of a solvent-
water mixture is defined as aw=
γw.xw where γw is the activity
coefficient and xw is the mole
fraction of the solvent
• The water activity can affect the
rate and selectivity of reaction of
the solid form that crystallises from
solution, i.e. whether it hydrated or
anhydrous

Bell et al. Enz Micr Tech 20: 471-477 (1997)

 Anti-solvent Crystallisation

• Determine the solubility of the

compound in a range of solvents at HCl salt
different temperatures(preferably ICH
Solubility of AZD1208 HCl salt
class II and (preferably class III) and
classify them as solvents or Solubility (mg/mL) measured at
Solvent (Class) 20 C 40 C 60 C
antisolvents Methanol (II) 350 385 ND
Ethanol (III) 60 63 70
• Test if they will crystallise the IPA (III) 10 12 15
n-Butanol (III) 12 13 16
compound by adding the antisolvent 2-MeTHF (IV) 0.0 0.0 0.0
to a solution of the compound MEK (III) 0.0 0.0 0.1
IPAc (III) 0.0 0.0 0.0
• Note, e.g. oiling out, gelling etc DMSO (III) >1000 >1000 >1000
MTBE (II) 0.0 0.0 0.0
• Select those that give the best yield, Water (III) >100 >100 >100
Anisole (III) 0.0 0.0 0.0
crystals etc
 Antisolvent Crystallisation -
Normal Addition Mode
Normal addition-
Antisolvent added to a
D solution containing
Solution Concentration of Product

the compound of
A interest
B
B’ Labile Zone • A-B Undersaturated zone
• B-B’ Metastable zone width
• D-E Metastable limit –no
Undersaturated zone

crystallisation , but will vary

depending stirring rate,
E antisolvent addition rate etc
• B’-C Desupersaturation to
Metastable C
solubility curve
zone

Quantity of Antisolvent Added

 Antisolvent-Reverse Addition
• The solubility is very low at point A
• Because of this low solubility, the Reverse addition – Solution
supersaturation ratio will increase rapidly containing the solvent
added to the antisolvent
(E-F) before sufficient seed surface area is

Solution concentration of Product

attained to achieve any growth
• If addition is sufficiently fast, growth will
F
significantly reduced resulting in Labile Zone
D
nucleation dominating over growth
• Small particles will often result B

• Small particles have the tendency to

agglomerate C
• May also cause oiling out, i.e. liquid-liquid A

de-mixing E Quantity of Product Solution Added

• Large drops of oil may coalescence,
won’t disperse and the harden into a
gummy/waxy solid
• Wax may occlude solvent and
impurities
 Problems Associated With Anti-Solvent
Crystallisations

 More difficult to control

 Increased risk for oil formation
 Gel formation
 Local high supersaturation at addition point
 fast crystallisation
 Incrustation at dip pipe

 small crystals gelators can be nanometer- or

micrometer-sized
 metastable forms can appear Particles, which forms 3D
network with solvents
 Varying solvent conditions during solvent addition
 metastable forms or mixture of forms

 solvate formation and transformation

 In general more difficult to scale-up than cooling crystallisation

Yin et al. Ind @Eng Chem Res 53 1286-1292 (2013)

 Limitations with Anti-Solvent Crystallisations

 Process aspects
 larger solvent volumes
 larger volume variation
 more difficult to recover-mixed solvents
 may require pump for control

Added volume of solvent

Time
 Possibilities With Anti-Solvent Crystallisations
• Flexibility
• Greater possibility to choose solvent system
• Improved purity by combination of polar and non-polar
systems
• Clear filtration steps are more easy to perform
• Not necessary to pre-heat pipes. etc
• Process economics
• Higher yields
• Less degradation or by-products
• Low temperature crystallisation possible
• Low temperature screening possible
 Considerations in Processes with Anti-Solvent
Addition
• The antisolvent addition may be applied alone or in combination
with other crystallization procedures. Some common examples are:
• Antisolvent addition at constant temperature, with or
without seeding
• In combination with cooling.
• At the start temperature
• Somewhere during cooling
• At the end temperature
 Optimum Antisolvent Addition Rate

• By design, antisolvent addition reduces the solubility of the solute in solution.

• The rate of antisolvent addition influences the supersaturation level and
hence the amount of growth and nucleation.
• Careful control of the addition rate can help control the size and number of
crystals produced.
• Slow initial rate to match slow rate of solid deposition.
• As surface area increases, mass deposition rate increases.
• Addition rate can be increased to match the crystallization rate.
 Optimum Antisolvent Addition Rate

• Typically antisolvent added a solution of the compound at a constant rate

• Slow addition tends to be better than fast addition
• Fast addition can result in poor mixing and high local supersaturations
• This can result in small particles due to uncontrolled crystallisation and the
generation of the wrong polymorphic form - especially if unseeded

H
N OH

.HCl

H
N
AZD9056 Crude (1)
MW 455.47
O Cl

1. MeOH/H2O, reflux
2. TBME, 50-55°C
3. 20-25°C

H
N OH

.HCl

H
N AZD9056 Pure (1)
MW 455.47
O Cl

Fast (immobile slurry) slow

 Optimum Antisolvent Addition Rate
• For better results, the antisolvent should be slowly added to the
solution at high rate of mixing.
• If seeds are used, it is advantageous to add the seeds to the
antisolvent first and then slowly introduce the suspension
(antisolvent plus the seeds) to the solution.
• This will prevent formation of regions of high supersaturations at
local and bulk levels, and therefore, excessive nucleation and
deterioration of crystal habit/size/polymorph will be avoided

See Nonoyama et al.

Org Proc Res Dev 10 727-732 (2006)
 Example - When Addition of Antisolvent?

 Compound is crystallised from MIBK/Heptane by cooling from

60°C and addition of antisolvent:
1. Heptane added at 60°C
2. Heptane added at 25°C
 The filtration time was decreased by applying procedure 2

Procedure 1 Procedure 2
 Seeding
• As with cooling crystallisations, seeding may help to avoid excessive nucleation
and help control the crystallisation
• The seed can be added as a slurry in the antisolvent (low solubility)
• Need to know when to add the seed
– Too early and it will dissolve (solution undersaturated) and too late nucleation will already
have taken place
• Therefore, need to know the seeding point!
• Can add near the saturation point whereby the antisolvent reduces the
solubility into the metastable zone and seeds will grow
• Or add some antisolvent to create some supersaturation and then add the
seeds in a slurry
• Typical to have a hold time of around 30 mins after seed addition to allow it
properly disperse and to take up the supersaturation
• The goal is to stay in the metastable zone so that growth the dominant process
 Antisolvent Crystallisation-Seeding
• System without seed or high
addition rate can develop
high supersaturation resulting
D in rapid precipitation or crash
out at B’’ (may lead to oiling
Solution Concentration of Product

out, small crystals,

A aggregation)
B
B’ B’’ • The closer the solution
Seeding
Zone concentration profile is to
F follow the equilibrium
solubility curve (B-C), the
Undersaturated zone

higher the possibility of

achieving an all growth
E process
Metastable C • Best route is B’-F-C with
Labile Zone
zone seeding at point B’
• Important that seed holds! So
Quantity of Antisolvent Added add some antisolvent to
create some supersaturation
• Add seed as a slurry
 AZD1208

HCl salt

System chosen was MeOH

(solvent), IPAc (antisolvent)

1. Dissolve AZD1208 API in 3 RV MeOH at 40C

2. Clear filter
3. Add 3 RV IPAc in 1.5h
4. Add seed (small/large seed with seed loading 0.1
to 1%w/w)
5. Hold for 0.5h
6. Add 9 RV IPAc in 4 to 12h
7. Hold for 10h
8. Filter
9. Wash with 10 RV IPAc
10. Suck dry for 10 min
11. Dry under vacuum (Pmin) at 65C for 24h
AZD1208HCl
AZD1208HCl
 Combined Cooling/Antisolvent Crystallisation
• As illustrated, antisolvent crystallisationis often
combined with cooling and it is common to add
the antisolvent before the cooling profile has
commenced or at it is added after the cooling
profile has finished
• However, this can have a profound effect on the
crystallistion of some compounds and in many
cases may not optimal from crystallisation
perspective
• Lindenberg et al. (2009) have optimised this

supersaturation
type of process and instead of adding the
antisolvent at the beginning and the end the
antisolvent is added during the cooling profile
with a view to maintaining constant
supersaturation.

Lindenberg et al.
Cryst Growth Des 9 1124-1136 (2009)
Time (s)
 Combined Cooling/Antisolvent Crystallisation
• Temperature and
antisolvent additions
are obviously non-
linear
• Might be difficult to
implement in an
industrial setting.
• Therefore, a constant
cooling and
antisolvent addition
regime was
examined and whilst
suboptimal, it was a
better strategy than
adding the
antisolvent at the
beginning or the end
of the cooling ramp.
 Combined Cooling/Anti-solvent Crystallisation
• Similar study has been conducted by Nagy et al. (2008) with lovastatin
• Similar non-linear cooling and antisolvent addition profiles were modelled
and implemented
• This resulted in significantly higher crystal quality and better yield than the
cooling only crystallisation (seeded was, of course, better than the unseeded
system)
Seed Seeded, Cooling only
Temperature (°C)

40

30

20

0 50 100 150 200

Time (min)
10
Flow rate (g/min)

0
0 50 100 150 200
Time (min)

Nagy et al. J Cryst Growth (2008) Antisolvent only Combined antisolvent/ cooling
 Combined Antisolvent/Cooling
Oiling Out
• A Bristol-Myers Squibb (BMS) drug
substance candidate oiled out during
an antisolvent process
• When cyclohexane was added rapidly
to a solution of the compound in ethyl
acetate it oiled out (they wanted small
crystals to obviate milling)
• They avoided oiling out by using an
initial solvent composition of 2:1
volume ratio by keeping the overall
polarity in the mid-region whilst
inducing nucleation from seeds by slow
cooling
• Once the nucleation started, cooling
was stopped to allow the crystallisation
to progress at low supersaturation.
• Could also be performed with ethanol- Kim et al. Org Proc Res Dev 7 997-1001 (2003)
water (preferred)
 Remacemide HCl Manufacture-Solvent Swap
• The recrystallisation remacemide HCl Final manufacturing process for remacemide hydrochloride

on scale was carried out as follows.

O
• A 3000 L glass-lined reactor was MgCl stage 1

+ THF OH
charged with the crude, IPA damp,
KCN, H2SO4
remacemide hydrochloride Stage 2
n-Bu2O
(equivalent to 275 kg dry weight) via
the manway. Stage 3
• Methanol (approximately 1100 L) was 1. Aqueous HCl
NH
NH2 2. NaOH/DCM H
charged to the reactor and the DPAP
O

mixture brought to reflux. The Me3COCl, BOC-Glycine

Stage 4
Et3N, DCM
solution obtained was pumped to a IPA, aqueous HCl HN
O
Stage 5
second 3000 L glass lined reactor via O NH2.HCl

2 m and 0.2 m filters. N

H N
H
O
crude remacemide HCl

• The vessel and filters were rinsed O Stage 6

recrystallisation IPA/methanol

through with more methanol (55 L). Pure remacemide HCl

 Remacemide HCl Manufacture
• The methanol solution was concentrated by distillation (approximately
275 L removed) and then IPA (approximately 1650 L) was charged to the
2nd reactor via a 0.2 μm filter.
• Distillation was continued to remove the required quantity of mixed
solvents (approximately 1705 L).
• IPA (approximately 550 L) was charged to the reactor via a 0.2 mm filter.
• Distillation was continued to remove the required quantity of mixed
solvents (approximately 550 L).
• The mixture was cooled to 0-5 oC and the product isolated, washed with
IPA (~275 L) and then dried in a stainless steel filter drier..
Anti-Solvent Addition: Scale-Up/Scale-Down
Issues

Problem with scale up…

Mixing!

Circulation times in the production

vessel are significantly longer than
those encountered in the
laboratory.
 Mixing
• The hydrodynamics of a stirred tank varies from loca-tion to
location,
• Mixing time scale analysis is essential for understanding and
controlling antisolvent (and reactive) crystallisation
processes
• Can be broken down into three distinct regions:
• Macromixing, mesomixing and micromixing
Macromixing Mesomixing Micromixing

Largest scale of mixing Describes the course-scale Since micromixing happens on

present in STRs and is a turbulent exchange between the molecular scale, it directly
function of agitation a fresh feed plume and its affects the chemical reaction
speed. surrounding environment. (and antisolvent mixing) and
It controls global mixing The antisolvent addition thus nucleation and growth,
times and mean rate, feed pipe diameter, which are molecular-based
concentration fields. location and agitation all processes
play a role in mesomixing.
 Relationship Between Mixing Scale and Mixing
Time.
• The mixing time can be loosely
defined as the time required to
reach 95% homogeneity of
multiple streams on mixing
• Since mixing of these streams
creates superaturation, the time
of mixing to reach composition
homogeneity will clearly affect
the subsequent nucleation and
crystal growth rates
• Mixing time is affected by
multiple factors, e.g. mixing
intensity, scale and mixer
geometry
Micro Scale - from μL to mL
Mesoscale – from mL to L
Macroscale – from L to 1000’s L
Tung Org Proc Res Dev 17 445-454 (2013)
 AntiSolvent Addition in the Lab

Antisolvent addition line

Small, well-mixed • Mixing is of the order of 1

vessel sec in lab reactors whereas
in large reactors it can be in
the order of ten seconds
Can lead differences in mass/heat
No or few transfer
concentration
gradients
 Anti-Solvent Addition in the Plant

Antisolvent addition line

• Concentration gradient, which
is not well distributed throughout
the vessel, it is present at the
surface, meaning:
Long circulation
time – Local high supersaturation
– Localised nucleation (which was
not seen in the well-mixed
Vessel not environment of the lab)
homogenous – The process does not scale well!
 Mixing

• Product size distribution becomes significantly influenced by the mixing

conditions
• The hydrodynamic situation in a turbulent STR is complex and can vary
significantly from location to location
• Mixing is due to the transport of the energy provided by the agitator
through a cascade of vortices to the smallest scales where the energy is
dissipated as heat
 Where to Add the Antisolvent?
Feed Location

At Above Above
Below
Impeller Impeller Surface
Impeller

Feed Rate Power Distribution

Low Fast Fast

Rate Multi-feed Single Single Large
Rate Impeller Impeller D/T Ratio
Rate
No Baffles With Baffles
 Where to Add the Antisolvent?
• A local high supersaturation may be created at the antisolvent addition
location if mixing is insufficient
• This may depend on where the antisolvent is added
• For example, near the wall or closer to the impeller
• Near the wall results in poorer mixing compared to addition to near the
impeller and the faster the addition rate the poorer the mixing
• Thus with incomplete mixing will get bulk primary nucleation + local
primary nucleation, whereas complete mixing will result in bulk primary
nucleation only
Mesomixing time scale (s)

Poorly mixed region

Well-mixed
region

FBRM Fine Counts/#s

Feedback Control

Zhou et al. Cryst Growth Des 6 892-898 (2006)

 Feedback Control
• Antisolvent rate addition rate
increases as the crystallisation Process is
seeded
progresses due to the o the increase
in the crystal surface area
• Unlike to T control (where it can
increase or decrease to maintain
constant supersatuation), once
added the antisolvent cannot be
removed.
• Therefore, to maintain constant
supersaturation solvent may need to
be added to the solution
• The obvious drawback of this is that
dilution and the number of process
volumes will increase
• Should be done at low
supersaturation to avoid excessive
secondary nucleation Zhou et al. Cryst Growth Des 6 892-898 (2006)
 Guanosine 5’ Monophosphate (GMP)
• Various concentrations of GMP in water were guanosine 5’ monophosphate (GMP

charged to a 0.5 L reactor to which methanol

was injected to precipitate the GMP.
• Initial precipitate was amorphous, which
subsequently transformed to a 7-hydrate 1.0

0.8
during the course of the addition.

of GMP hydrate
Mass fraction
0.6

• The phase transformation rate was found to 0.4

be promoted by increasing the agitation rate 0.2 200 rpm, 61 g/L feed concentration
600rpm 122 g/L feed concentration
(better mass transfer), 0.0
600 rpm 30.5 g/L feed concentration

• Too high a agitation led to attrition with a 0 50 100 150 200

Crystallisation time (min)
250 300

concomitant decrease in particle size. 200

• Similarly, the concentration of the GMP in 150

Mean crystal size (μm)

solution influenced the transformation and
100
growth of the crystals.
50
• Up to certain concentration growth was 200 rpm, 61 g/L feed concentration
0 600rpm 122 g/L feed concentration
enhanced, however, past this critical point a 600 rpm 30.5 g/L feed concentration
0 100 200 300 400
change in morphology and secondary Crystallisation time (min)

nucleation took place. Kang et al. Appl Biochem Biotechnol 160 561-573 (2010)
 Naftazone
• In precipitation processes, it is the nucleation step that determines the
polymorphic form that is produced, i.e. if JII > JI then II will form will form
rather than I.
• If the nucleation rates are similar then mixes can occur leading to so-called
concomitant polymorphs
• Addition of the antisolvent (water) to a solution of 1,2-Naphthoquinone-2-
semicarbazone (naftazone) at high concentration (30 g L-1) in DMF
resulted in the production of the stable form of the compound even at
the highest addition rate of the antisolvent.
• At a lower concentration (15 g L-1) at slow addition rate Form II
(metastable) was obtained. At intermediate concentrations and water
additions concomitant crystallisation of the polymorphs.

Dureisseix et al.
Cryst Growth Des 9 3438-3443 (2009)
. Crystallisation of an Intermediate

• Slow addition of water gave bright yellow needles of 4 in >93% isolated

yield.
• Unfortunately, these crystals filtered exceptionally poorly, typically requiring
24 h filtration time, and leading to difficulties in removing both reaction by-
products and residual solvents.

H2O

Org Proc Res Dev, 10, 751 – 756 (2006)

 Crystallisation of an Intermediate

• Crystallisation from alternative solvents did not give improvement

• However, inverting order of addition gave soft agglomerates of fine
needles
• Hoever, agitation can reduce particle size
• Scaled up using pitched blade agitator
• Reduced filtration time from 24h to 90 min

Org Proc Res Dev, 10, 751 – 756 (2006)

 Agglomeration / Spherical Crystallisation
• The crystals agglomerate and form spherical particles during or after
the crystallisation, e.g. aspirin
• Choice of the three solvents; good solvent – ethanol; anti-solvent –
water; bridging liquid – toluene (small amount, immiscible with water)
• Heated aspirin in ethanol to 60 ºC, added toluene into the drug
solution.
• Poured the drug solution with toluene into the reactor containing
water. Stirring speed 450 rpm.

Precipitated crystals cluster around droplets of

the bridging liquid
 Agglomeration / Spherical Crystallisation

Ethanol

Water Toluene

Some problems with residual

solvents
 Impinging Jets

• Microfluidics Reaction Technology

(MRT) machines are based on high
shear and high pressure impinging jet
technology using solvent/anti-solvent
streams through fixed geometry
capillary chambers (residence times 1
ms).
• MRT has appears to have the greatest
nanoparticle stability, narrowest
distribution, and up to 60% solids
concentration

configured with coaxial feed for longer residence times to

accommodate a slower crystallization process
 Impinging Jets
• Microfluidics Pure Nano
Continuous Crystalliser is being
used to create stable inhalable
formulations, improve the
bioavailability of antibiotics and
target delivery of novel cancer
treatments
• This type of equipment could be
scaled up, e.g. inside the
microfluidiser production units,
identical microchannels are
aligned in parallel.
• Isolation of solid without filtration
is possible, by e.g. continuous
spray drying, prilling, granulation,
scraped heat exchanger etc.
 Summary
• Next most popular choice to cooling crystallisation
• More difficult to control and can many limitations , e.g. oiling out gelling,
small crystals, wrong polymorph
• Has some advantages, e.g. low temperature crystallisation
• Like cooling crystallisation can improve control by seeding
• Feedback control possible, but more result in too many volumes!
• Spherical agglomeration of crystals possible