processes

Article
Systematic Design and Evaluation of an
Extraction Process for Traditionally Used
Herbal Medicine on the Example of Hawthorn
(Crataegus monogyna JACQ.)
Maximilian Sixt and Jochen Strube *
Institute for Separation and Process Technology, Clausthal University of Technology,
38678 Clausthal-Zellerfeld, Germany; sixt@itv.tu-clausthal.de
* Correspondence: strube@itv.tu-clausthal.de; Tel.: +49-05-323722200




Received: 29 May 2018; Accepted: 19 June 2018; Published: 21 June 2018


Abstract: Traditionally used herbal medicines are deep in the consciousness of patients for the
treatment of only minor diseases by self-medication. However, manufacturers of herbal medicinal
products suffer from major problems such as increasing market pressure by e.g., the food supplement
sector, increasing regulations, and costs of production. Moreover, due to more stringent regulation
and approval processes, innovation is hardly observed, and the methods used in process development
are outdated. Therefore, this study aims to provide an approach based on modern process engineering
concepts and including predictive process modelling and simulation for the extraction of traditional
herbal medicines as complex extracts. The commonly used solvent-based percolation is critically
assessed and compared to the so-called pressurized hot water extraction (PHWE) as a new possible
alternative to replace organic solvents. In the study a systematic process design for the extraction
of hawthorn (Crataegus monogyna JACQ.) is shown. While for traditional percolation the solvent
is optimized to a mixture of ethanol and water (70/30 v/v), the PHWE is run at a temperature of
90 ◦ C. The extracts of various harvest batches are compared to a commercially available product
based on a chromatographic fingerprint. For the first time, natural batch variability was successfully
incorporated into the physico-chemical process modelling concept. An economic feasibility study
reveals that the PHWE is the best choice not only from a technical point of view but also from
economic aspects.

Keywords: pressurized hot water extraction; modelling; simulation; harvest; variety

1. Introduction
Herbal medicine is widely used to cure a myriad of diseases such as gastrointestinal complaints,
high blood pressure, sore throat, colds, skin disorders and other sicknesses. They are considered to be
natural, compatible and healthy. On account of its long tradition, the use and production of herbal
medicine must not be overtaken by modern engineering technologies for production and quality
management. Assessing small-molecule pharmaceuticals approved between 1981 and 2002 shows
that approximately half are either natural products or natural product derivatives [1,2]. In the future,
natural products will continue to play a major role as active substances, model molecules for the
discovery and validation of drug targets [1].
Therefore, this paper highlights the systematic process development, simulation and cost
assessment for a commonly used herbal drug represented by hawthorn (Crataegus monogyna JACQ.).
The conventional solvent extraction is compared with the so-called pressurized hot water extraction
(PHWE) [3–6] which shows significant potential for the eco-friendly and solvent-free extraction of

Processes 2018, 6, 73; doi:10.3390/pr6070073 www.mdpi.com/journal/processes

Processes 2018, 6, 73 2 of 19

plant matrices [3,4,6]. Hawthorn leaves are a representative system for a medical extract. It is used to
treat cardiac insufficiency [7]. The flavonoid hyperoside is chosen as a marker substance. The extracts
will be characterized by their content of hyperoside and the drug-extract ratio (DER). The DER is
defined as the ratio between the plant material used for extraction and the amount of dry matter
remaining in the fluid extract after the extraction process has taken place, according to Equation (1).
mPlant
DER = : 1 (1)
mDry matter

Corresponding monographs in the European Pharmacopoeia define no specific DER for hawthorn
extracts [8]. An overview of the drug market shows that common products are extracted within a
range of DER values between 4–7:1 [9–13]. This range is therefore taken as a benchmark.

2. Material and Methods

2.1. Raw Material

Raw hawthorn leaves (Crataegus monogyna JACQ. folium cum flore) were purchased from CfM
Oskar Tropitzsch (Markredwitz, Germany). The herb was ground to approximately 1 mm size prior to
extraction in a knife mill Retsch® Grindomix® 200 (Haan, Germany). The plant material was stored
in the refrigerator at −20 ◦ C to minimize decomposition. Pure hyperoside was purchased from CfM
Oskar Tropitzsch (Markredwitz, Germany), as well. The purity was >98%.

2.2. Solvent Screening

Solvent screening is performed in 50 mL Falcons® (VWR® , Darmstadt, Germany) using 3 g of
ground plant material and 40 mL of solvent. The mixtures are agitated on a tilting table for 24 h before
a sample is taken and analyzed. All solvents are in analytical grade.

2.3. Solid-Liquid Extraction

The conventional extraction is performed as maceration for the equilibrium measurements
and as percolation for the determination of the extraction kinetics as well as the overall amount.
The maceration device is a stirred tank of approximately 1.5 L. It is equipped with a three-way top
were the joint in the middle is used to insert a stirrer, driven by a lab stirrer motor from IKA® (Staufen,
Germany). A PTFE seal prevents solvent from leaking through the gap between the top and the vessel.
The device can be tempered. Percolation is performed in a stainless steel tube equipped with frits
to ensure proper solid retention during extraction. A P110 preparative HPLC pump from VWR®
as well as a Foxy Jr. fraction collector are used. Both are controlled by the EZChrom Elite® HPLC
Software (Agilent® , Santa Clara, CA, USA). The experimental setups have already been described by
the authors [14,15].

2.4. Pressurized Hot Water Extraction (PHWE)

The flow diagram of the PHWE equipment for batch extraction experiments is shown in Figure 1.
The water is pumped from a storage vessel by a HPLC pump P130 from VWR® (Darmstadt, Germany)
to the extraction column filled with ground plant material. The extraction column is made from
stainless steel with 10 mm i.d. × 100 mm (approx. 7.85 mL). The column itself and a coil made of a steel
capillary (length = 3 m, i.d. = 1 mm) are placed in a GC oven HP 5890 Series II Plus (Palo Alto, CA, USA)
serving as heating device. The water is preheated in the coil before reaching the extraction column.
The extract leaves the extraction column at the back-end where a filter frit serves for solid-liquid
separation. The extract is transferred into a second coil for effective cooling (length = 3 m, i.d. = 1 mm).
The pressure drop is adjusted by a valve to keep the water in liquid state.
Processes 2018, 6, 73
Processes 3 of 19
Processes2018,
2018,6,6,xxFOR
FORPEER
PEERREVIEW
REVIEW 33 of
of 19
19

Data
DataLogger
Logger
TT TT
PP
PP
Valve
Valve

Pump
Pump Heater
Heater Extraction
ExtractionVessel
Vessel Cooler
Cooler
GC-Oven
GC-Oven

Storage
StorageVessel
Vessel Extract
Extract

Figure 1. Flow
Figure diagram ofofthe milli-scale PHWE equipment for
for batchextraction
extraction experiments.
Figure1.
1.Flow
Flowdiagram
diagram ofthe
themilli-scale
milli-scalePHWE
PHWEequipment
equipment forbatch
batch extractionexperiments.
experiments.

ToTo
To determine
determine
determinethe the thermal
thermal degradation
thethermal degradation of
degradation ofofthethe
the investigated
investigated
investigated components
components
components during extraction,
during
during the
extraction,
extraction, the
themilli-scale
milli-scaledevice
milli-scale device
device is modified
is is modified
modified to
to operate
to operate
operate in
in recycling
in recycling
recycling mode.
mode. This
This setup
mode. This is
is shown
setupsetup in
in Figure
is shown
shown 2.
2. The
in Figure
Figure The 2.
Theextract
extract
extractis
is transferred
transferred
is transferred into the
the storage
intointo storage
the storagevessel
vessel (max. 100
100 mL)
(max.(max.
vessel mL) and
and
100 is
mL)is recycled
recycled during
duringthe
and is recycled the whole
wholethe
during process.
process.
whole
Thus,
process. the
Thus, Thus, target
the target component
component
the target accumulates
accumulates
component in the system
in the system
accumulates and occurring
and occurring
in the system degradation
degradation
and occurring effects
effects can
degradation can be
be
effects
studied.
canstudied.
be studied.The storage
The Thestorage vessel
vessel
storage itself is
itselfitself
vessel in
is in an
an an
is in ultrasonic
ultrasonic
ultrasonic bath
bath
bathfor mixing
forformixing and
mixingand avoiding
andavoiding possible
avoidingpossible
possible
agglomerates
agglomerates as well as concentration gradients inside the vessel.
agglomerates as as well
well as as concentrationgradients
concentration gradientsinsideinsidethethe vessel.
vessel.

Data
DataLogger
Logger
TT TT
PP
PP
Valve
Valve

Pump Heater
Heater Extraction
ExtractionVessel
Vessel Cooler
Pump Cooler
GC-Oven
GC-Oven

Storage
StorageVessel
Vessel
Ultrasonic
UltrasonicBath
Bath

Figure
Figure 2. 2.2.
Figure FlowFlow
Flow diagram
diagramofof
diagram the
ofthe milli-scale
themilli-scale PHWEequipment
milli-scalePHWE equipmentin
equipment inrecycling
in recyclingmode.
recycling mode.
mode.

Both setups have already been introduced by the authors [5,6].

Both setups have already beenintroduced
introducedby
bythe
theauthors
authors [5,6].
[5,6].
Both setups have already been
2.5. Analytics
2.5.2.5. Analytics
Analytics
The analytics for hyperoside in the extract is performed by HPLC using an Elite LaChrom ®®
TheThe analytics
analytics forfor hyperosideininthe
hyperoside theextract
extract isis performed
performed by by HPLC
HPLC using
using ananElite
EliteLaChrom
LaChrom®
(Agilent® ,, Santa Clara, CA, USA) device equipped with aa diode array detector DAD-L 2455 from
(Agilent®,® Santa Clara, CA, USA) device equipped with a diode® array detector DAD-L 2455from
(Agilent® Santa Clara, CA, USA) device equipped with diode array detector DAD-L 2455 from
Hitachi
Hitachi
® ® (Tokyo, Japan). The eluents are in analytical grade (VWR , Darmstadt,
(Tokyo, Japan). The eluents are in analytical grade (VWR® , Darmstadt, Germany) and water
® Germany) and water
Hitachi (Tokyo, Japan). The eluents are in analytical grade (VWR , Darmstadt, Germany) and water
is
is taken
taken from
from aa Sartorius
Sartorius
®® arium®® pro (Göttingen, Germany). All samples are passed through a 0.2
arium® pro (Göttingen, Germany). All samples are passed through a 0.2
® arium
is taken
µµm
from a Sartorius pro (Göttingen,performed
Germany). All samples are passed through a
m syringe
syringe filter
filter prior
prior to
to analysis.
analysis. Calibration
Calibration was
was performed with with external
external standards
standards ordered
ordered fromfrom
0.2CfM
µm syringe filter prior to analysis. Calibration was performed with external standards ordered
CfM Oskar
Oskar Tropitzsch
Tropitzsch (Marktredwitz,
(Marktredwitz, Germany).
Germany). The The purity
purity was
was greater
greater 98%.
98%. The
The column
column was was aa
from
250-4CfM Oskar Tropitzsch (Marktredwitz, Germany).Germany),
The purity was greater 98%. The column
250-4 mmmm PharmPrep
PharmPrep RP18 RP18 from
from Merck
Merck® (Darmstadt, Germany), the the column
column temperature
temperature is is 25
25 °C
®
(Darmstadt,
® (Darmstadt, Germany), the column temperature is
°C
wasthea 250-4
flow mm
rate PharmPrep
is 1 mL/min RP18
and from
the Merck
isocratic eluent consists of methanol/acetonitrile/water (pH 2.5
the flow rate is 1 mL/min and the isocratic eluent consists of methanol/acetonitrile/water (pH 2.5
◦ C the flow rate is 1 mL/min and the isocratic eluent consists of methanol/acetonitrile/water
25 with
with ortho-phosphoric
ortho-phosphoric acid) acid) 16/21/63
16/21/63 v/v/v.
v/v/v. The
The detection
detection wavelength
wavelength was was 355
355 nm
nm and
and the
the injection
injection
(pH 2.5
volume with 5 ortho-phosphoric
µ L. The chromatogram acid) 16/21/63
is shown v/v/v.
in Figure The3.detection
The
volume 5 µ L. The chromatogram is shown in Figure 3. The coefficient of determination wavelength
coefficient of was 355
determinationnmof and
of thethe
the
injection volume 5 µL. The chromatogram is shown in Figure 3. The coefficient of determination of the
Processes 2018, 6, 73 4 of 19

Processes 2018, 6, x FOR PEER REVIEW 4 of 19

calibration was 0.99 and the limit of quantification 0.02 g/L. The side components (SCs) SC1 to SC3 are
calibration was 0.99 and the limit of quantification 0.02 g/L. The side components (SCs) SC1 to SC3
referred to the calibration of hyperoside and expressed as pseudo-concentrations. IR-spectroscopy
are referred to the calibration of hyperoside and expressed as pseudo-concentrations.
was performed with
IR-spectroscopy a FTIR
was Bruker®with
performed alphaa (Billerica,
FTIR BrukerMA,® USA).
alpha Principal
(Billerica,Component
MA, USA).Analysis was
Principal
calculated using Unscrambler ® X from Camo® (Oslo, Norway).
Component Analysis was calculated using Unscrambler X from Camo (Oslo, Norway).
® ®

300

250
Absorption (mAU)

200

150 SC1
Hyperoside
100
SC3
50

0 SC2
0 1 2 3 4 5 6 7 8 9 10
Time (min)

Figure3.3.Chromatogram
Figure Chromatogram of
of the
the hawthorn
hawthornextract.
extract.

3. Results
3. Results andand Discussion
Discussion

Remark. Every experiment is carried out in triplicates. The error bars represent the standard deviation of the
Remark. Every experiment
measurements. is carried
If no error bars out they
are visible in triplicates.
are withinThe error bars
the symbol sizerepresent the standard
of the individual deviation of the
data points.
measurements. If no error bars are visible they are within the symbol size of the individual data points.
The scope of this article is to introduce the reader to a modern process engineering attempt for
theThemanufacturing of traditionally
scope of this article used the
is to introduce herbal
readerextracts including
to a modern physico-chemical
process engineering attempt processfor
themodelling and experimental
manufacturing of traditionallyparameter determination
used herbal on lab-scale.
extracts including The benefits of
physico-chemical the modelling
process modelling
andand simulation parameter
experimental approach for phytoextraction
determination are documented
on lab-scale. in detail
The benefits of thebymodelling
various studies of the
and simulation
authors [5,6,14–23]. The cited articles focused mostly on the extraction of valuable
approach for phytoextraction are documented in detail by various studies of the authors [5,6,14–23]. compounds for
plants in order to perform a further purification until a pure substance in pharma-grad
The cited articles focused mostly on the extraction of valuable compounds for plants in order to perform is obtained,
e.g., forpurification
a further 10-deacetylbaccatin
until a pureIII substance
from yew in [6,18] or artemisinin
pharma-grad from annual
is obtained, e.g., formugworth [5]. In the III
10-deacetylbaccatin
present
from study,or
yew [6,18] the engineering
artemisinin fromtoolbox
annualwill be broadened
mugworth [5]. Intothe
thepresent
processingstudy,of complex extractstoolbox
the engineering that
are characterized by marker substances and the DER in a regulated environment, within the area of
will be broadened to the processing of complex extracts that are characterized by marker substances
conflict of being economical on the one side and fulfilling approvals on the other side. Therefore,
and the DER in a regulated environment, within the area of conflict of being economical on the one
modern and in-silico-based approaches are needed, especially if there is a new approval of existing
side and fulfilling approvals on the other side. Therefore, modern and in-silico-based approaches are
processes considered to broaden the area of application or to just modernize the production
needed, especially if there is a new approval of existing processes considered to broaden the area of
procedure.
application or to just modernize the production procedure.
In general, solid-liquid extraction is described by several partial models. The corresponding
In general, solid-liquid can
equations and assumptions extraction
be found is in
described by literature
detail in the several partial models.
[14,15,17]. The massThebalance
corresponding
in the
equations and assumptions can be found in detail in the literature [14,15,17].
liquid phase is described by the so-called distributed plug flow model (DPF). It serves for The mass balance
modelingin the
liquid phase is described
the macroscopic by the so-called
mass transport distributedcolumn.
in the percolation plug flow Formodel (DPF).
the solid It serves
phase a porefordiffusion
modeling
themodel
macroscopic
describes the mass transport in the plant material’s pores. The raw material’s residual diffusion
mass transport in the percolation column. For the solid phase a pore load of
model describes
target components the mass
and thetransport in the plant
corresponding material’s
extract’s pores. is
concentration The raw material’s
accounted residual load
for by equilibrium
of curves.
target components
The target and the corresponding
component’s extract’s
mass balance in concentration
the fluid phase is accounted
(Equationfor(2)) by equilibrium
considers
curves. The targetaxial
accumulation, component’s
dispersion, mass balance inmass
convective the fluid phase (Equation
transport and mass (2)) considers
transfer fromaccumulation,
the plant
material’s
axial poresconvective
dispersion, into the bulk
mass phase.
transport and mass transfer from the plant material’s pores into the
bulk phase.
∂cL (z,t) ∂2 cL (z,t) uz ∂cL (z,t) 1−ε
= Dax ⋅ − ⋅ − ⋅ kf ⋅ aP ⋅ [cL (z,t) − cP (r,z,t)] (2)
∂t ∂z 2 ε ∂z ε
∂cL (z, t) 2
∂ cL (z, t) uz ∂c (z, t) 1−ε
= Dax · − · L − · kf · aP · [cL (z, t) − cP (r, z, t)] (2)
∂t ∂z2 ε ∂z ε
Processes 2018, 6, 73 5 of 19

Under the assumption of spherical particulates, the following equation can be derived as a mass
balance for the target component in the plant material. The effective diffusion coefficient Deff must be
measured by real extraction experiments.
Processes 2018, 6, x FOR PEER REVIEW 5 of 19
∂2 cP (z, r, t)
 
∂q(z, r, t) 2 ∂cP (z, r, t) ∂Deff (r) ∂cP (z, r, t)
= Deff (r) · + · + · (3)
Under ∂r2 particulates,
∂tthe assumption of spherical r the following
∂r equation∂r
can be derived
∂r as a mass
balance for the target component in the plant material. The effective diffusion coefficient D eff must be
The adsorption/desorption
measured equilibrium inside the pores is described through equilibrium curves.
by real extraction experiments.
Herein q is the plant material’s residual2 load linking the equilibrium curve with the pore diffusion
∂q(z,r,t) ∂ cP (z,r,t) 2 ∂cP (z,r,t) ∂Deff (r) ∂cP (z,r,t)
model. A well-known approach (r)⋅ (the Langmuir-equilibrium.
= Diseffe.g., + ⋅ ) + The ⋅ KL and q(3)
∂t ∂r2 r ∂r ∂r parameters
∂r max can be
derived from measurements. The maximum load qmax is the total amount of the regarded target and
The adsorption/desorption equilibrium inside the pores is described through equilibrium
side components. The equilibrium curves are measured with multistep macerations. The approach is
curves. Herein q is the plant material’s residual load linking the equilibrium curve with the pore
described in detail
diffusion in the
model. literature approach
A well-known [14,15]. is e.g., the Langmuir-equilibrium. The parameters KL and
qmax can be derived from measurements. The maximum load qmax is the total amount of the regarded
KL · c
q = qmax
target and side components. The equilibrium · are
curves measured with multistep macerations. The (4)
approach is described in detail in the literature [14,15]. KL · c
1 +

3.1. Solvent Screening KL ⋅ c

q = qmax ⋅ (3)
1 + KL ⋅ c
To achieve a high extraction performance, solvent selection is crucial. Because hawthorn extract
is used3.1.
as Solvent
a traditional
Screeningherbal medicine, not every solvent that is ideal from an engineering point of
view is suitable. Commonly solvents or their aqueous solutions which are described in monographs
To achieve a high extraction performance, solvent selection is crucial. Because hawthorn extract
are used. According
is used to theherbal
as a traditional European Pharmacopeia,
medicine, water
not every solvent that isorideal
ethanol
from an water solutions
engineering with
point of more
than 45view
vol.is % of ethanol
suitable. are used
Commonly fororthe
solvents production
their of a siccum
aqueous solutions which are that is afterwards
described processed to
in monographs
a dry extract
are used.[8]. To provide
According to thea European
realistic study, ethanolwater
Pharmacopeia, wateror solutions
ethanol water ranging from
solutions pure
with morewater to
than 45 vol. % of ethanol are used for the production of a siccum that is
pure ethanol in 10 vol. % steps are screened. The results are presented in Figure 4. The greyafterwards processed to a section
dry extract [8]. To provide a realistic study, ethanol water solutions ranging from pure water to pure
contains ethanol water solutions that are not reported in the European Pharmacopeia, as depicted
ethanol in 10 vol. % steps are screened. The results are presented in Figure 4. The grey section
earlier. Besides hyperoside three other side components are regarded in the screening. They are
contains ethanol water solutions that are not reported in the European Pharmacopeia, as depicted
referred to hyperoside
earlier. by pseudo-concentrations.
Besides hyperoside A solution
three other side components of 70 vol.
are regarded % ethanol
in the rest
screening. water
They are shows
the highest
referred to hyperoside by pseudo-concentrations. A solution of 70 vol. % ethanol rest water shows for the
capacity towards the target molecule hyperoside. Therefore, it is chosen as solvent
conventional process
the highest design.
capacity towardsBoth, puremolecule
the target water and pure ethanol
hyperoside. have
Therefore, it issignificantly lower
chosen as solvent for capacities
the
towards conventional
hyperoside process design.mixtures
than their Both, pure waterMoreover,
have. and pure ethanol
they are have
at significantly
approximately lowerthe
capacities
same level of
relativetowards hyperoside than their mixtures have. Moreover, they are at approximately the same level of
solubility. This indicates that screening for completely different pure solvents, such as e.g.,
relative solubility. This indicates that screening for completely different pure solvents, such as e.g.,
n-propanol or methanol, is not sufficient but aqueous solutions have to be considered.
n-propanol or methanol, is not sufficient but aqueous solutions have to be considered.

Figure 4. Solvent screening for hyperoside from hawthorn in ethanol water solutions.
Figure 4. Solvent screening for hyperoside from hawthorn in ethanol water solutions.
Processes 2018, 6, x FOR PEER REVIEW 6 of 19
Processes 2018, 6, 73 6 of 19
According to the screening following solvent compositions are preferred:
• Hyperoside,
According to 70
thevol. % ethanol,
screening rest water
following solvent compositions are preferred:
• SC1, 30 vol. % ethanol, rest water
•• Hyperoside,
SC2, 30–40 vol. 70 vol. % ethanol,
% ethanol, rest water
rest water
•• SC1, 30 vol. % ethanol, rest
SC3, 70 vol. % ethanol, rest water water
• SC2, 30–40 vol. % ethanol, rest water
Probably SC1 to SC3 are flavonoids as well, because their adsorption maxima in
• SC3, 70 vol. % ethanol, rest water
UV/VIS-spectra show significant bands at 207–215 nm, 257–270 nm and 332–359 nm [24].
Probably
Established SC1 to SC3
products are are flavonoids
commonly as well,with
extracted because their%adsorption
45 vol. ethanol, rest maxima
waterin[9–13].
UV/VIS-spectra
Assuming
show
SC1 tosignificant
SC3 are alsobands at 207–215
flavonoids, thisnm, 257–270
solvent is anm and 332–359
preferable mixturenmto [24]. Established
achieve high totalproducts
flavonoid are
commonly extracted
content in the extract. with 45 vol. % ethanol, rest water [9–13]. Assuming SC1 to SC3 are also flavonoids,
this solvent is a preferable mixture to achieve high total flavonoid content in the extract.
3.2. Model Parameter Determination for Conventional Extraction
3.2. Model Parameter Determination for Conventional Extraction
For proper modeling and simulation, the model parameters equilibrium and overall amount
mustForbeproper modeling
determined. Theandoverall
simulation,amount the model parameters
of hyperoside is equilibrium
derived byand overall amount
exhaustive must
percolation
be determined.
experiments Theethanol/water
using overall amount of hyperoside
(70/30 v/v). It is is derived
0.35% by exhaustive
± 0.02% referred to percolation
dry mass.experiments
The overall
using ethanol/water (70/30 v/v). It is 0.35% ± 0.02% referred
amounts of SC1 to SC3 regarding hyperoside are 0.82%, 0.19% and 0.35%, respectively. to dry mass. The overall Inamounts
literatureof
SC1 to SC3
values regarding
for the overall hyperoside
flavonoid amountare 0.82%, 0.19%1–2%
between and 0.35%, respectively.
are given [7]. In literature values for the
overall flavonoid amount between 1–2% are given [7].
The equilibrium data was measured by multi step maceration experiments and is shown in
FigureThe
5. equilibrium data was
The Langmuir-type measured equilibrium
distribution by multi step formaceration
hyperosideexperiments
and SC3 areand bothisrather
shownflat, in
Figure 5. The
confirming theLangmuir-type
chosen ethanol distribution
water mixture equilibrium
as well-chosenfor hyperoside
solvent. The and SC3 are both
equilibrium rather
curves flat,
for SC1
confirming the chosen ethanol water mixture as well-chosen solvent.
and SC2 are almost approaching to their specific overall amount, on the contrary. There is a The equilibrium curves for SC1
and SC2 are
relatively almost
high approaching
amount of thesetocomponents
their specificremaining
overall amount,
in the on the matrix,
plant contrary.soThere is a relatively
the solvent is not
high amount of these components remaining in the plant matrix, so the solvent
ideally chosen for these components. This finding is consistent to the solvent screening of Section 3.1. is not ideally chosen
for these5 components.
Figure also containsThis the finding
equilibriumis consistent to the of
relationship solvent screening
the whole dry of Section
matter 3.1. Figure
extracted from 5 also
the
contains
hawthorntheleaves.
equilibrium relationship
This parameter is offurther
the whole neededdry matter extracted
for proper from the
modeling hawthorn
of the leaves.
DER during
This parameter
extraction. is further
The overall needed
amount for matter
of dry properwas modeling
determinedof thetoDER
34%,during
thus theextraction.
minimum The DERoverall
would
amount
be 2.9:1. of dry matter was determined to 34%, thus the minimum DER would be 2.9:1.

300
A B
250

200
Residue load (g/l)

150

100

50

0
0 10 20 30 40
Concentration (g/l)

Figure 5. (A) Equilibrium data for the system hawthorn-hyperoside/SC1/SC2/SC3-EtOH/Water

Figure 5. (A) Equilibrium data for the system hawthorn-hyperoside/SC1/SC2/SC3-EtOH/Water
(70/30 v/v); (B) equilibrium data for the system hawthorn-dry matter-EtOH/Water (70/30 v/v).
(70/30 v/v); (B) equilibrium data for the system hawthorn-dry matter-EtOH/Water (70/30 v/v).

3.3. Model Parameter Determination for PHWE

3.3. Model Parameter Determination for PHWE
For modeling and simulation of the PHWE process a suitable extraction temperature must be
For modeling and simulation of the PHWE process a suitable extraction temperature must be
defined. Therefore, a temperature screening was performed ranging from 80 ◦°C to 140 ◦°C in 20 ◦°C
defined. Therefore, a temperature screening was performed ranging from 80 C to 140 C in 20 C
Processes 2018, 6, 73 7 of 19
Processes 2018, 6, x FOR PEER REVIEW 7 of 19

steps. Afterwards
Afterwardsthe theindividual degradation
individual kinetics
degradation are determined.
kinetics The pressure
are determined. was keptwas
The pressure constant
kept
to 15 bars in all experiments, to ensure the best possible comparability.
constant to 15 bars in all experiments, to ensure the best possible comparability.

3.3.1. Extraction Temperature

3.3.1. Extraction Temperature
The
The results
results of
of the
the screening
screening are
are presented
presented in
in Figure
Figure 6.
6.

100% 100%
90% 90%
80% 80%
70% 70% 80 °C
100 °C
60% Hyperoside 60%

Yield
Yield

120 °C
50% 50% 140 °C
40% 40%
80 °C
30% 30%
100 °C
20% 120 °C 20%
SC1
10% 140 °C 10%
0% 0%
0 20 40 60 80 100 120 0 20 40 60 80 100 120
Extraction time (min) Extraction time (min)

100% 100%
90% 90%
80% 80%
70% 70% 80 °C
60% 60% 100 °C
Yield

Yield

120 °C
50% 50%
140 °C
40% 40%
80 °C
30% 30%
100 °C
20% SC2 120 °C 20%
10% 140 °C 10% SC3
0% 0%
0 20 40 60 80 100 120 0 20 40 60 80 100 120
Extraction time (min) Extraction time (min)

Figure 6. Temperature screening.

Figure 6. Temperature screening.

• Hyperoside: The screening shows that hyperoside is almost entirely extracted after 60 min at
• Hyperoside:
100 °C. A Yield Theofscreening showsafter
95% is reached thatabout
hyperoside
20 minisatalmost
120 °Centirely
and 140extracted after
°C. At that 60 min at
temperature,
100 ◦ C. A Yield ofis95% is reached after aboutto20100
min°Catand ◦
120 700%
C andhigher◦
140 C. At that temperature,
the productivity 250% higher compared compared to 80 °C. A
the productivity is 250% higher compared to 100 ◦ C and 700% higher compared to 80 ◦ C.
decline in yield with rising temperature cannot be observed, indicating a high thermal stability
A
of decline in yield with rising temperature cannot be observed, indicating a high thermal stability
hyperoside.
• of hyperoside.
Side component 1: The productivity of extraction is nearly constant in the regarded
• Side component
temperature range The productivity
1: from 80 °C to 140 of°C.extraction
Thermal is nearly
decay constant
does in the
not seem regardedastemperature
to appear, well.
◦ C to 140 ◦ C. Thermal decay does not seem to appear, as well.
• range from 80
Side component 2: SC2 shows the greatest dependency on temperature among the regarded
• Side component
substances. At 80 2:°CSC2 shows
a yield the greatest
of only about 70%dependency on temperature
is reached after among
120 min. This yield the regarded
is reached at
substances.
140 °C after 80 ◦ C
Atjust 15a yield
min. ofTheonly about 70%
increase is reached after
in productivity is 120 min. This yield
approximately is reached
800%. at
Thermal
degradation cannot be traced based on this data.
Processes 2018, 6, 73 8 of 19

◦ C after just 15 min. The increase in productivity is approximately 800%.

1402018, Thermal degradation
Processes 6, x FOR PEER REVIEW 8 of 19
cannot be traced based on this data.
•• Side component 3: 3: SC3 shows
shows high productivity
productivity among the whole temperature field. Besides
that, the yields at 80 ◦°C ◦ C are under the level of 100 ◦ C (90% yield after 120 min). At a
C and 140 °C °C
temperature of 140 ◦°C
temperature C this can indicate thermal decomposition, which is investigated
decomposition, which investigated in the
◦ C the productivity is limited because the temperature is not well-chosen
following section.
section.AtAt80 80 °C the productivity is limited because the temperature is not
for this substance.
well-chosen for this substance.
Already commercialized hawthorn
Already commercialized hawthorn drugs
drugsare
areextracted
extractedtotoa specific
a specific DER
DER ranging
ranging from
from 4:1 4:1 to
to 7:1.
7:1. To ensure a good comparability towards established products, that parameter is investigated
To ensure a good comparability towards established products, that parameter is investigated as well. as
well.
FigureFigure
7 shows7 the
shows the extraction
extraction curves forand
curves for hyperoside hyperoside
the DER inand the DER intemperature
the investigated the investigated
range.
temperature range.

10 80 °C 90 °C
100 °C 120 °C 100%
9
140 °C 90%
8
80%
7
70%
6
60%
DER

Yield

5
50%
4
40% 80 °C
3 90 °C
30%
2 100 °C
20% 120 °C
1 10% 140 °C
0 0%
A 0 20 40 60 80 100 120 B 0 20 40 60 80 100 120
Extraction time (min) Extraction time (min)

Figure 7.
Figure (A) DER
7. (A) DER during
during the
the PHWE
PHWE of
of hawthorn;
hawthorn; (B)
(B) Extraction curve of hyperoside during PHWE.

At
At 8080 °C◦ C aayield
yieldofof85%85% regarding
regarding hyperoside
hyperoside is reached
is reached afterafter
120 120
min minof extraction. The
of extraction.
corresponding
The corresponding DERDER is approximately
is approximately 7.2:1.7.2:1.
In that casecase
In that onlyonlya small
a small amount
amountof dry extract
of dry is
extract
obtained that contains a relatively high quantity of hyperoside, so the extraction
is obtained that contains a relatively high quantity of hyperoside, so the extraction is rather selective. is rather selective.
By
By rising
rising temperature
temperaturealmost almostcomplete
completeextraction
extractionofof hyperoside
hyperoside is is
achieved.
achieved. At the same
At the time,time,
same the
DER decreases
the DER decreasesto values of 3.5:1
to values at 100
of 3.5:1 °C, 2.7:1
at 100 at 120
◦ C, 2.7:1 °C and
at 120 2:1 at
◦ C and 2:1140 °C, ◦being
at 140 smaller
C, being thanthan
smaller the
minimum value obtained at conventional percolation with ethanol/water.
the minimum value obtained at conventional percolation with ethanol/water. None of the examined None of the examined
temperature
temperature matches
matches aa high
high yield
yield of
of hyperoside
hyperoside and and aa DER
DER in in the
the desired
desired range
range at at the
the same
same time.
time.
Therefore, an extraction at 90 ◦ °C was performed. A yield of hyperoside of
Therefore, an extraction at 90 C was performed. A yield of hyperoside of 90% and a DER of 5.5:1 after 90% and a DER of 5.5:1
after
120 min120ofmin of extraction
extraction have beenhavemeasured,
been measured,
thus thisthus this temperature
temperature is chosenisforchosen
hot water for hot water
extraction
extraction
of hyperosideof hyperoside from hawthorn.
from hawthorn. In that
In that case, thecase, the process
process cannot cannot
be named be named pressurized
pressurized hot
hot water
water extraction by definition. From a process engineers point of view this is
extraction by definition. From a process engineers point of view this is not decisive, because the shown not decisive, because
the shown
model model determination
parameter parameter determination
concept doesconcept doesand
not change not moreover,
change and themoreover,
extractionthe extraction
is now is
possible
now possible
without without
additional additional
pressure, thuspressure,
reducingthus reducing
investment investment
and operatingand costsoperating
at the samecosts at the same
time.
time.
3.3.2. Thermal Degradation
3.3.2. Thermal Degradation
To be aware of thermal degradation in hot water extraction is the key parameter for successful
To
extractionbe [5,6].
awareAs ofshown
thermalindegradation
the screening, in no
hotclear
waterthermal
extraction
decay is the
couldkeybeparameter
traced byfor thesuccessful
provided
extraction
data. For [5,6].
that, As shown in the experiments
recycling-mode screening, nowere clear performed
thermal decay at 120could◦ C be
andtraced
140 by◦ C the provided
to assess the
data.
impact Forofthat,
highrecycling-mode
temperature toward experiments were performed
hyperoside. The resultsatare 120given
°C and 140 °C to
in Figure 8. assess
As it can thebeimpact
seen,
of high temperature toward hyperoside. The results are given in Figure 8. As it can be seen, all
regarded components are quite stable at high temperatures. Only SC3 shows a rather small decline
in concentration after 4 h of process time. At the chosen extraction temperature of 90 °C no thermal
Processes 2018, 6, 73 9 of 19

Processes 2018, 6, x FOR PEER REVIEW 9 of 19

all regarded components are quite stable at high temperatures. Only SC3 shows a rather small decline
in concentration
decay after
occurs, and the4 degradation
h of process time. At the
kinetics canchosen extraction
be neglected temperature
in the of 90 ◦ C no
physico-chemical thermal
extraction
decay occurs, and the degradation kinetics can be neglected in the physico-chemical extraction model.
model.

0.10

0.09 SC1
0.08

0.07
Concentration (g/l)

0.06 140°C
Hyperoside 120°C
0.05

0.04
SC3
0.03

0.02 SC2

0.01

0.00
0 25 50 75 100 125 150 175 200 225 250
Process time (min)

Figure 8. Concentration during the recycling experiment for all respective substances.
Figure 8. Concentration during the recycling experiment for all respective substances.

3.4. Evaluation of Hot Water Extraction and Solvent-Based Percolation

3.4. Evaluation of Hot Water Extraction and Solvent-Based Percolation
Experimental data (points) as well as the simulation results (drawn lines) of the conventional
Experimental data (points) as well as the simulation results (drawn lines) of the conventional
percolation of hawthorn are shown in Figure 9. Between 80 and 180 min of extraction, the acceptable
percolation of hawthorn are shown in Figure 9. Between 80 and 180 min of extraction, the acceptable
range of the DER between 7:1 and 4:1 is exceeded. The extraction curves of the components show a
range of the DER between 7:1 and 4:1 is exceeded. The extraction curves of the components show
strong bend in this timespan and according to economic considerations, the extraction should be
a strong bend in this timespan and according to economic considerations, the extraction should be
aborted after 250 min at the latest. The DER shows a strong correlation with the individual
aborted after 250 min at the latest. The DER shows a strong correlation with the individual substances,
substances, as expected. In that case the DER is a suitable tool for a basic process control, besides
as expected. In that case the DER is a suitable tool for a basic process control, besides being technically
being technically overtaken by various procedures of in-line monitoring ranging from
overtaken by various procedures of in-line monitoring ranging from Raman-spectroscopy [20,25],
Raman-spectroscopy [20,25], IR-spectroscopy [25] or simple conductivity measurement. Because the
IR-spectroscopy [25] or simple conductivity measurement. Because the simulation of the pressurized
simulation of the pressurized hot water extraction simplifies to a percolation, due to the thermal
hot water extraction simplifies to a percolation, due to the thermal decay not being considered,
decay not being considered, the approach of modeling and model parameter determination is
the approach of modeling and model parameter determination is similar to that already shown and
similar to that already shown and therefore is not shown in detail once again.
therefore is not shown in detail once again.
Processes 2018, 6, 73 10 of 19
Processes 2018, 6, x FOR PEER REVIEW 10 of 19

10

0.12
DER 9
SC1

0.1
8

0.08 7
Mass (g)

Hyperoside

DER
6
0.06

SC3
5

0.04
4

0.02
3

SC2
0 2
0 100 200 300 400 500 600 700 800
Extraction time (min)

Figure 9. Conventional extraction of hawthorn, experiments and simulation for hyperoside and side
Figure 9. Conventional extraction of hawthorn, experiments and simulation for hyperoside and side
components SC1 to SC3.
components SC1 to SC3.
Besides process engineering aspects, the extract composition plays an important role. For an
Besides process
evaluation extracts engineering aspects, the
of both, percolation andextract
PHWEcomposition
with a DERplays an were
of 6:1 important role. to
compared Fora an
evaluation extracts
commercially of both,product
available percolation and45
(ethanol PHWE with
vol. %, a DER
DER of 6:1
4–7:1). were compared
Assessed to a commercially
are the ratios of the side
components
available relative
product to hyperoside.
(ethanol The
45 vol. %, chromatograms
DER are shown
4–7:1). Assessed are thein Figure
ratios 10
of and are normalized
the side components
to the peak
relative of hyperoside.
to hyperoside. The chromatograms are shown in Figure 10 and are normalized to the
peak of hyperoside.
Processes 2018, 6, 73 11 of 19
Processes 2018, 6, x FOR PEER REVIEW 11 of 19

1200

1000
Adsorption (mAU)

800 SC1 PHWE 90°C

Percolation
600
Dry extract
400 Hyperosid SC3
e
200 SC2

0
1 2 3 4 5 6 7 8
Time (min)

Figure 10. Chromatogram of the hawthorn extract and exemplified finger print region.
Figure 10. Chromatogram of the hawthorn extract and exemplified finger print region.
• Side component 1: SC1 shows a minor content in percolation compared to the commercially
• Side component
available SC1 showstoathe
1: According
product. minor content
solvent in percolation
screening, compared
SC1 dissolves better intothe
thesolvent
commercially
of the
available product.
reference According
product. to the solvent
A significantly higher screening,
amount ofSC1 dissolves
SC1 betterduring
is extracted in the solvent of the
hot water
extraction.
reference product. A significantly higher amount of SC1 is extracted during hot water extraction.
• • Side
Side component2:2:SC2
component SC2shows
shows the
the same
samebehavior
behaviorasasSC1. According
SC1. Accordingto the solvent
to the screening,
solvent it
screening,
dissolves better
it dissolves betterin
inthe
thesolvent
solventused
usedtotoproduce
produce the reference
the extract.
reference extract.
• • Side component 3: The relative amount of SC3 in the referenceextract
Side component 3: The relative amount of SC3 in the reference extractisissmaller
smallerthan
thanin in
thethe
percolation performed in this study. According to the solvent screening the used mixture of 70
percolation performed in this study. According to the solvent screening the used mixture of
vol. % ethanol and 30 vol. % water has a much higher capacity towards this substance
70 vol. % ethanol and 30 vol. % water has a much higher capacity towards this substance
compared to the solution of the reference product. Again, the PHWE delivers an even higher
compared to the solution of the reference product. Again, the PHWE delivers an even
amount.
higher amount.
The chromatograms match the observations of the solvent screening. The PHWE reaches the
The chromatograms
highest values of all sidematch the observations
components, therefore itof the
can besolvent
consideredscreening. The PHWE
as less selective, reachestothe
compared
highest values of all
the percolation. side components,
It must be consideredtherefore it can be extract
that the reference considered as less
was not selective,
obtained fromcompared
the same to
thebatch of plant material
percolation. It must used for the shown
be considered that extraction experiments
the reference of this
extract was notstudy, but the
obtained fromplace
theand
same
yearofofplant
batch harvest are the used
material same.for
Moreover,
the shown it isextraction
not the objective of the shown
experiments of thisfingerprint
study, butapproach,
the placeto and
evaluate the quality or efficacy of the extracts, besides it is not yet known, which
year of harvest are the same. Moreover, it is not the objective of the shown fingerprint approach, substances or group
to of substances
evaluate the are responsible
quality for itof
or efficacy [26].
theProducts
extracts,that are only
besides it isspecified
not yet by the DER,
known, commonly
which pass or
substances
an internal
group qualityare
of substances management
responsibleprocess, where
for it [26]. fingerprint
Products methods
that are are widely
only specified by used [8]. With
the DER, the
commonly
shown approach of experimental model parameter determination on lab scale and
pass an internal quality management process, where fingerprint methods are widely used [8]. With the physico-chemical
modeling, it is now possible to predictively simulate markers, which are monitored in fingerprint
shown approach of experimental model parameter determination on lab scale and physico-chemical
methods, anyway, while monitoring the DER at the same time. Advanced process control and a
modeling, it is now possible to predictively simulate markers, which are monitored in fingerprint
model-based endpoint decision are possible [20]. This requires a process development with regards
methods, anyway, while monitoring the DER at the same time. Advanced process control and a
to the principles of Quality-by-Design (QbD) in a new approval of the product [19].
model-based endpoint decision are possible [20]. This requires a process development with regards to
the3.5.
principles of Quality-by-Design (QbD) in a new approval of the product [19].
Batch Variablity
A fundamental
3.5. Batch Variablity challenge in extracting plants is the content of the ingredients, which fluctuates
due to natural growth and site conditions. In terms of a model description of the solid-liquid
A fundamental challenge in extracting plants is the content of the ingredients, which fluctuates
extraction, the total content and the equilibrium per batch would have to be re-measured, if no
due to natural growth and site conditions. In terms of a model description of the solid-liquid extraction,
Processes 2018, 6, 73 12 of 19

Processes 2018, 6, x FOR PEER REVIEW 12 of 19

the total content and the equilibrium per batch would have to be re-measured, if no systematics can be
derived between
systematics can bethe two parameters.
derived between theTo determine
two a possible
parameters. dependency,
To determine a total
a possible of nine different
dependency, a total
batches, which differ by crop year (and thus storage life) and location, were examined. The
of nine different batches, which differ by crop year (and thus storage life) and location, wereequilibrium
data are shown
examined. in Figure 11data
The equilibrium and are
are interpreted as Langmuir-isotherms.
shown in Figure 11 and are interpreted as Langmuir-isotherms.

0.50
0.45
0.40
0.35
Residue load (g/l)

0.30 A
0.25
C
0.20 E
D
0.15 I
B
0.10
G
0.05 H
0.00 F

0.0 0.1 0.2 0.3 0.4 0.5

Concentration (g/l)

Figure
Figure 11.11. Equilibria
Equilibria of harvest
of different different
batches harvest batches
of hawthorn of hawthorn-hyperoside-EtOH/Water
in the system hawthorn in the system
hawthorn-hyperoside-EtOH/Water
(70/30 v/v). (70/30 v/v).

The Langmuir coefficient is considered constant for all curves. The maximum load q max is
The Langmuir coefficient is considered constant for all curves. The maximum load q is basically
basically determined by the total content of the respective component. Experimentally,maxhowever, it is
determined by the total content of the respective component. Experimentally, however, it is observed
observed that the equilibrium practically never reaches this final value. The deviation is described
that the equilibrium practically never reaches this final value. The deviation is described based on a
based on a model parameter a, which is referred to below as the capacity factor according to
model parameter a, which is referred to below as the capacity factor according to Equation (5).
Equation (5).
KLL ⋅ ·c c
K
q · ⋅aa · ⋅
q =q q=max (5)
(4)
max + KKL L⋅ c· c
11 +
Obviously,
Obviously, ininthe the equilibrium
equilibrium relationships,
relationships, the maximum
the maximum loading loading is not reached,
is not reached, which
which indicates
indicates
that there that
is nothere
pure ad-is no or pure ad- orequilibrium
desorption desorption in equilibrium
the extraction in the extraction
of plants. A highof plants.
partitionAofhigh the
partition of the valuable substance is dissolved e.g., in the vacuole or in
valuable substance is dissolved e.g., in the vacuole or in cell interspaces. This corresponds to the model cell interspaces. This
corresponds
concept ‘Broken to theandmodel
Intactconcept
Cells’ ‘Broken
by Sovová and[27].
Intact Cells’ by
Kassing Sovová
et al. have[27].
takenKassing
up this etconcept
al. have in taken
the
up
porethis conceptmodel
diffusion in theand pore diffusion model
implemented it usingand implemented
a radial it using adistribution
target component radial target[14,15].
component If the
distribution
target component[14,15]. If the target
distribution cancomponent
be measured distribution
by methodscan suchbe asmeasured
Raman mappingby methods [25], such
this can as
Raman mapping If[25],
be implemented. this can be implemented.
no corresponding data are available, If nothecorresponding
introduction ofdata are available,
the capacity factor as thea
introduction of the capacity factor as a macroscopic parameter reflects the
macroscopic parameter reflects the basic idea of the ”Broken and Intact Cells” model, which is based basic idea of the ”Broken
and Intact Cells” model,
on discontinuously which is based
differentiable on discontinuously
equilibrium lines [28,29]. differentiable
If the solvent equilibrium
can dissolvelines [28,29]. If
and transport
the
away solvent
a largecan dissolve
amount andtarget
of the transport away aorlarge
substances, amount
if it is of the
accessible, onlytarget substances,
a small fraction,or if it is
which
accessible, only a small fraction, which is actually adsorbed in the cell or
actually adsorbed in the cell or between the cells, remains. An extreme example is the fennel fruit, between the cells, remains.
An extreme
in which theexample
essential is oilthe fennel fruit,
is localized in whicharound
in channels the essential oil is
the actual localized
fruit and in in channels
principle around
only has tothe be
actual fruitbyand
displaced thein principle
solvent, whichonlyleads
has to
to be
verydisplaced by themeasurable
flat or barely solvent, which leads tolines
equilibrium very[30].
flat or
If, barely
on the
measurable
other hand, theequilibrium
solvent has lines [30].
only If, oncapacity,
a small the other hand,
then much theofsolvent has only
the dissolved a small capacity,
substance remains inthen the
much of the dissolved substance remains in the cell or a large amount of solvent must be made
available. In this case, clearly pronounced equilibrium lines result.
The resulting equilibrium lines for the respective systems hyperoside-hawthorn-ethanol/water
(70/30 v/v) are shown in Figure 11. The Langmuir coefficient was chosen to be the same for all
datasets (75). There are differences only in the measured total content and in the capacity factor. The
Processes 2018, 6, 73 13 of 19

cell or a large amount of solvent must be made available. In this case, clearly pronounced equilibrium
lines result.
The resulting equilibrium lines for the respective systems hyperoside-hawthorn-ethanol/water
Processesv/v)
(70/30 2018, are shown
6, x FOR inREVIEW
PEER Figure 11. The Langmuir coefficient was chosen to be the same for all datasets
13 of 19
(75). There are differences only in the measured total content and in the capacity factor. The total
content, year and
total content, yearcountry of harvest
and country and the
of harvest capacity
and factor factor
the capacity a are summarized in Tablein1.Table
a are summarized Batch1.I Batch
is the
reference batch used
I is the reference inused
batch the previous experiments.
in the previous experiments.

Table 1. Overview of the hawthorn batches.

Overall Deviation to Deviation to

Batch Harvest
Harvest Year Origin Overall Deviation to Deviation to
Batch Origin Amount Reference a a Reference
Year Amount Reference Reference
A 2017 Southeast Europe 0.87% 146% 0.11 −15%
A 2017 Southeast Europe 0.87% 146% 0.11 −15%
BB 2016
2016 Macedonia
Macedonia 0.35%
0.35% −1% −1% 0.10
0.10 −−23%
23%
CC 2017
2017 Bulgaria
Bulgaria 0.58%
0.58% 64% 64% 0.12
0.12 −−8%
8%
DD 2017
2017 Romania
Romania 0.41%
0.41% 16%16% 0.130.13 0%
0%
EE 2014
2014 Bulgaria
Bulgaria 0.60%
0.60% 71% 71% 0.10
0.10 −−23%
23%
FF 2017
2017 Albania
Albania 0.42%
0.42% 20% 20% 0.020.02 −−84%
84%
G 2017 Southeast Europe 0.57% 62% 0.03 −77%
G 2017 Southeast Europe 0.57% 62% 0.03 −77%
H 2017 Serbia 0.51% 43% 0.015 −88%
H
I (Reference) 2017
2017 Serbia
Germany 0.51%
0.35% -43% 0.015
0.13 −88%
-
I (Reference) 2017 Germany 0.35% - 0.13 -

The
The overall
overall amount
amount with
with respect
respect to
to the
the reference
referencebatch
batchIIvaries from−−1%
variesfrom 1% to
to +146%. The capacity
+146%. The capacity
factor
factor assumes values between 0.10 and 0.13 for all batches except F to G. Batches F to G showed
assumes values between 0.10 and 0.13 for all batches except F to G. Batches F to G showed
significantly fewerflowers
significantly fewer flowersininthe
the extraction
extraction material;
material; thethe capacity
capacity factorfactor for these
for these samples
samples was
was 0.021
0.021 ± The
± 0.006. 0.006. The results
results are shown
are shown graphically
graphically in Figure
in Figure 12. 12.

1.0 0.2
Overall amount
0.9 0.18
Capacity factor
0.8 0.16
Overall amount (%)

0.7 0.14
0.6 0.12
a [-]

0.5 0.1
0.4 0.08
0.3 0.06
0.2 0.04
0.1 0.02
0.0 0
A B C D E F G H I
Batch

Figure 12. Evaluation of the batch screening regarding the overall amount and the capacity factor.
Figure 12. Evaluation of the batch screening regarding the overall amount and the capacity factor.

Obviously, the capacity factor does not correlate with the total content. It is, therefore, not
Obviously,Since
batch-specific. the itcapacity factor in
is understood does
the not
modelcorrelate
conceptwith
as a the total content.
macroscopic It is, therefore,
implementation of the
not batch-specific. Since it is understood in the model concept as a macroscopic
Broken and Intact Cell model, the data support this hypothesis. In addition, batches F, G and H show implementation
of the Brokenwhich
a parameter and Intact Cell model,
is clearly thefrom
different data the
support
otherthis hypothesis.
batches In addition,
and is very probablybatches
relatedF, G
to and
the
H show a parameter which is clearly different from the other batches and is very probably
lower share of flowers in the extraction material. The comminution of the extraction material here related to
the lower share of flowers in the extraction material. The comminution of the extraction
apparently causes a different, reproducible cell disruption, which consequently results in a different material here
apparently causes
capacity factor. a different,
With the same reproducible cell disruption,
pre-treatment and a visual which consequently
inspection results in
to determine thea different
share of
capacity factor. With the same pre-treatment and a visual inspection to determine
flowers in each batch, so only the total content must be measured. The capacity factor can then the share of flowersbe
assumed to be constant.
This data is the basis to simulate the individual percolations of the different batches, as shown
in Figure 13. The capacity factor is constant at 0.115 for batches A to E and I and 0.021 for batches F,
G and H. Apart from the individual total content of the respective batch, no further value is changed
in the simulation.
Processes 2018, 6, 73 14 of 19

in each batch, so only the total content must be measured. The capacity factor can then be assumed to
be constant.
This data is the basis to simulate the individual percolations of the different batches, as shown in
Figure 13. The capacity factor is constant at 0.115 for batches A to E and I and 0.021 for batches F, G
and H. Apart from the individual total content of the respective batch, no further value is changed in
the simulation.
Processes 2018, 6, x FOR PEER REVIEW 14 of 19

0.2
Batch A Batch B Batch C
0.18 Batch D Batch E Batch F
Batch G Batch H Batch I
0.16

0.14

0.12
Mass (g)

0.1

0.08

0.06

0.04

0.02

0
0 100 200 300 400
Time (min)

Figure 13. Simulation of the extraction of different hawthorn batches.

Figure 13. Simulation of the extraction of different hawthorn batches.

The deviations between simulation and experiment relative to the experimentally determined
total The
massdeviations between
of hyperoside simulation
are listed in Tableand experiment
2. The relative
simulations to the A
of batches experimentally determined
and B are underestimated
total
by 16%mass
andof10%,
hyperoside are listed
respectively. For in Table
these two2. batches,
The simulations of batches
the average Asize
particle and was
B areprobably
underestimated
smaller
by 16% and 10%, respectively. For these two batches, the average particle size was probably smaller
than expected, which limits the diffusion in the simulation more than was the case in the experiment.
than expected, which limits the diffusion in the simulation more than
The remaining batches have maximum deviations of +5% and 4%, respectively. was the case in the experiment.
The remaining batches have maximum deviations of +5% and 4%, respectively.
Table 2. Overview of the simulation results.
Table 2. Overview of the simulation results.
Batch Mass Experiment (g) Mass Simulation (g) Difference (g) Deviation
A Batch 0.135
Mass Experiment (g) 0.161
Mass Simulation (g) −0.026 (g)
Difference −16%
Deviation
B 0.054 0.060 −0.006 −10%
A 0.135 0.161 −0.026 −16%
C 0.090 0.091 −0.001 −1%
B 0.054 0.060 −0.006 −10%
D C 0.0610.090 0.063
0.091 −0.002
−0.001 −−3%
1%
E D 0.0940.061 0.090
0.063 0.004
− 0.002 5%
−3%
F E 0.0740.094 0.077
0.090 −0.003
0.004 −4%
5%
G F 0.0970.074 0.093
0.077 0.004
− 0.003 4%
−4%
H G 0.0870.097 0.093
0.086 0.004
0.000 4%
0%
H
I (Reference) 0.0560.087 0.086
0.054 0.000
0.002 0%
3%
I (Reference) 0.056 0.054 0.002 3%
Examination of individual hawthorn batches showed large fluctuations of the hyperoside
content. Nevertheless,
Examination the equilibrium
of individual hawthorncould be showed
batches described uniformly
large across
fluctuations of all
the batches.
hyperosideUsing this
content.
database, onlythe
Nevertheless, theequilibrium
total content of abe
could new batch must
described be measured
uniformly across alland a predictive
batches. simulation
Using this database, is
possible. In addition, the capacity factor can be correlated with complex matrix
only the total content of a new batch must be measured and a predictive simulation is possible. IR-spectra of the
powdered
In addition,plant material.factor
the capacity Thesecan
fingerprint spectra,
be correlated which
with are determined
complex as triplicates,
matrix IR-spectra of the are shown
powdered
in Figure
plant 14A. A
material. principal
These component
fingerprint analysis
spectra, which (PCA) of the second
are determined derivative
as triplicates, ofshown
are these spectra shows
in Figure 14A.
a detailed clustering of the different batches, as depicted in Figure 14B. The batches
A principal component analysis (PCA) of the second derivative of these spectra shows a detailed F, G and H with
the lower share of flowers are clearly separated from the batches with high share of flowers.
Therefore, the fast track data acquisition for predictive simulation of the lot variety can be performed
with IR-spectroscopy using the raw material.
Processes 2018, 6, 73 15 of 19

clustering of the different batches, as depicted in Figure 14B. The batches F, G and H with the lower
share of flowers are clearly separated from the batches with high share of flowers. Therefore, the fast
track data acquisition for predictive simulation of the lot variety can be performed with IR-spectroscopy
using
Processesthe raw
2018, 6, xmaterial.
FOR PEER REVIEW 15 of 19

0.002
0.06
High
Low
A 0.05 B
0.001

0.04

Intensity

PC-4
0.03 0

0.02
-0.001
0.01

0
-0.002
1400 1200 1000 800 600 400 -0.003 -0.002 -0.001 0 0.001 0.002 0.003
Wavenumber (cm-1) PC-1

Figure 14. (A) Spectra of the different harvest batches, black: batches F, G, H (low share of flowers),
Figure 14. (A) Spectra of the different harvest batches, black: batches F, G, H (low share of flowers), grey:
grey: rest (high share of flowers); (B) PCA of the second derivative of the hawthorn harvest batches
rest (high share of flowers); (B) PCA of the second derivative of the hawthorn harvest batches spectra.
spectra.

4. Economic Feasibility Study

4. Economic Feasibility Study
Both hot water extraction and conventional percolation have their own specific advantages and
Both hot water extraction and conventional percolation have their own specific advantages and
disadvantages. Based on a profitability analysis, economic aspects should be considered in addition to
disadvantages. Based on a profitability analysis, economic aspects should be considered in addition
the purely procedural aspects. To make the cost study, the following assumptions were made:
to the purely procedural aspects. To make the cost study, the following assumptions were made:
•• 30
30 tons
tons ofof hawthorn
hawthorn leavesleaves are
are to
to be
be extracted
extracted annually
annually in in 60
60 batches
batches on on aa multi-product
multi-product device.
device.
The
The purchase
purchaseprice priceisis3 3€/kg.
€/kg.The
Theannual
annualoccupancy
occupancy of of
thethe
system
system with thisthis
with product
productshould be 25%.
should be
• For the SLE as well as for the PHWE, an extractor of 2 m 3 empty volume is available.
25%.
• The
For investment
the SLE as costs well foras the
for SLE
the amount
PHWE, to an200,000
extractor€. The
of 2high-pressure
m3 empty volume equipment is 100% more
is available. The
expensive
investmentthan the
costs forconventional
the SLE amount technology and €.
to 200,000 amounts to 400,000 €. equipment is 100% more
The high-pressure
• Operating
expensive than parameters and process
the conventional times and
technology up amounts
to 95% to yield
400,000are €.directly taken from the
• laboratory studies.
Operating parameters and process times up to 95% yield are directly taken from the laboratory
• studies.
The energy costs are split into solvent preheating (here PHWE only) and evaporation. To operate
• Theevaporators
the energy costs and arethesplit into solvent
preheating (120 ◦ C, 5 bar,
steam preheating (here PHWE only)
2.7 MJ/kg) is usedand price of 13 €/t.
at aevaporation. To
• operate the evaporators and the preheating steam (120 °C, 5 bar, 2.7 MJ/kg)
10% of the solvent quantity per extraction is lost due to losses and must be renewed continuously. is used at a price of
13 addition,
In €/t. once a year, the entire pre-existing solvent, which is assumed to be 20 m , is replaced. 3

• 10%the
For of ethanol,
the solvent of 3 €/L per
a pricequantity and extraction
for water 0.08 €/L is
is lost due to losses and must be renewed
assumed.
• continuously. In addition, once a year, the entire
A worker is scheduled for the operation of the plant. The labor pre-existing solvent,
costs which is assumed
amount to 100,000to be€/a
20
m , isare
and3 replaced.
weighted For according
the ethanol, toa the
price of 3process
total €/L and time
for water
of the 0.08 €/L is assumed.
extraction of hawthorn added
• A worker is scheduled
(multi-product plant!). for the operation of the plant. The labor costs amount to 100,000 €/a and
• are costs
The weighted according and
for depreciation to the total process
maintenance time
for this of theamount
product extraction of of
to 2.5% hawthorn added
the total system
(multi-product plant!).
costs (multi-product investment!).
• The costs for depreciation and maintenance for this product amount to 2.5% of the total system
The
costsresult of the costinvestment!).
(multi-product study is shown in Figure 15. The total annual PHWE costs are about 50%
less than conventional extraction. The biggest difference is the annual solvent change. Hot water
The result of the cost study is shown in Figure 15. The total annual PHWE costs are about 50%
extraction can save about 99% of the costs compared to SLE. If the solvent is not renewed annually,
less than conventional extraction. The biggest difference is the annual solvent change. Hot water
the PHWE is still about 30% cheaper. However, the evaporation of the aqueous extract takes up about
extraction can save about 99% of the costs compared to SLE. If the solvent is not renewed annually,
10 times the amount of energy. The significantly shorter process time for hot water extraction means
the PHWE is still about 30% cheaper. However, the evaporation of the aqueous extract takes up
about 10 times the amount of energy. The significantly shorter process time for hot water extraction
means that staff can be used much more efficiently, which means that the personnel costs of this
product are around 20% lower than in the comparative process.
Processes 2018, 6, 73 16 of 19

that staff can be used much more efficiently, which means that the personnel costs of this product are
around 20% lower than in the comparative process.
Processes 2018, 6, x FOR PEER REVIEW 16 of 19

€300,000

€250,000 SLE
PHWE
€200,000
Costs / a

€150,000

€100,000

€50,000

€-

Figure 15. Comparison of the annual costs of the hawthorn extraction for conventional solid-liquid
Figure 15. Comparison of the annual costs of the hawthorn extraction for conventional solid-liquid
extraction and PHWE.
extraction and PHWE.

The cost calculation shows that the PHWE is competitive, even if the equipment is much more
The cost
expensive. In calculation
addition, itshows
can bethat
seenthe PHWE
that is competitive,
the cost even extraction
of conventional if the equipment is muchbymore
is dominated the
expensive. In addition, it can be seen that the cost of conventional extraction is
solvent. This means that often procedurally sensible operating points, here 70% ethanol and 30% dominated by the
solvent. This means that often procedurally sensible operating points, here 70%
water as solvent, are not feasible for economic reasons. In this case, this results in the ethanol and 30% water
as solvent, are not feasible
industry-preferred for economic
solvent mixture of onlyreasons. In thiswhich
45% ethanol, case, drastically
this resultsreduces
in the industry-preferred
the cost of solvent
solvent mixture of only 45% ethanol, which drastically reduces the cost
renewal but is not the optimum according to solvent screening. Here, PHWE offers of solvent renewal but is not
the opportunity
the optimum according to solvent screening. Here, PHWE
to implement the optimum process technology at reasonable costs. offers the opportunity to implement the
optimum process technology at reasonable costs.
5. Effort Analysis
5. Effort Analysis
The evaluation of hot water extraction and conventional percolation is based on a valid model
The evaluation of hot water extraction and conventional percolation is based on a valid model
parameter determination which can be handled under sensible experimental effort. For the
parameter determination which can be handled under sensible experimental effort. For the modelling
modelling and simulation of extraction processes to become routine industrial processes, the path
and simulation of extraction processes to become routine industrial processes, the path shown must be
shown must be superior to a purely empirical design using statistical experimental design. For this
superior to a purely empirical design using statistical experimental design. For this purpose, Figure 16
purpose, Figure 16 shows a schedule for a complete parameter determination which must be
shows a schedule for a complete parameter determination which must be handled by a person with
handled by a person with the corresponding equipment. Black is the actual working time, dark gray
the corresponding equipment. Black is the actual working time, dark gray are the times in which the
are the times in which the experiment does not have to be supervised and light gray is the time in
experiment does not have to be supervised and light gray is the time in which the analysis takes place.
which the analysis takes place.

Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10
Experiment
Solvent Screening
Equilibrium
Overall amount
Temperature Screening
Degradation Kinetics

Figure 16. Time schedule of the experimental setup.

 modelling and simulation of extraction processes to become routine industrial processes, the path
shown must be superior to a purely empirical design using statistical experimental design. For this
purpose, Figure 16 shows a schedule for a complete parameter determination which must be
handled by a person with the corresponding equipment. Black is the actual working time, dark gray
are the
Processes times
2018, 6, 73 in which the experiment does not have to be supervised and light gray is the time17in
of 19
which the analysis takes place.

Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10
Experiment
Solvent Screening
Equilibrium
Overall amount
Temperature Screening
Degradation Kinetics

Figure 16. Time schedule of the experimental setup.

Figure 16. Time schedule of the experimental setup.

For a complete parameter determination, with each assay being run as a triple determination,
appr. 10 days are required. The classical extraction experiments can be left unattended most of the
time, which is why the more labor-intensive model parameter determination for hot water extraction
can take place in parallel. To determine all parameters, only appr. 300 g of plant material and 8 L of
solvent are consumed. If a statistical test plan is used instead of model parameter determination and
simulation, one percolation can be carried out daily with the same equipment. Even with only three
parameters and one center point, a full-factorial experimental design results in 11 experiments, which,
taken together, have a much lower information density than the shown path of model parameter
determination and a predictive process simulation. The model-based approach is therefore to be
preferred in any case, especially since with increasing experience the results can be very quickly
checked for plausibility or can be transferred to other material systems.

6. Conclusions
The study showed a systematic and model-based comparison of two different ways of
manufacturing a traditionally used herbal extract. Both a percolation using a mixture of ethanol and
water (70/30 v/v) as solvent as well as an extraction with water at 90 ◦ C show high productivities and
yields. A high yield of the main flavonoid hyperoside as well as the desired range of the DER is reached.
The chromatographic fingerprints revealed that all extracts are comparable to a commercially available
one. The combination of experimental model parameter determination and rigorous process model is
an efficient method of predictive process simulation, not only for the extraction of substances which are
afterwards purified to pharma-grade, but also for the processing of traditionally used complex extracts.
For the first time, natural batch variability was successfully incorporated into the physico-chemical
process modelling concept. These generated data sets are required by regulatory authorities demanding
Quality-by-Design (QbD) and Process Analytical Technology (PAT) approaches as modern tools with
data-driven decisions documented for filing due to technological change, entering of markets of other
countries as well as changes of regional regulations and authority inquiry. An economic feasibility
study showed that the PHWE can overcome the financial drawbacks of solvent storage and renewal
efficiently, thereby justifying the higher investment costs for the necessary high-pressure equipment.

Author Contributions: M.S. did the conceptuation, experiments, simulations and validation. J.S. did the
supervision and the reviewing of the manuscript.
Funding: We also acknowledge the financial support obtained from the Deutsche Forschungsgemeinschaft (DFG)
in Bonn, Germany (project Str 586/4-2). The authors want to thank the Bundesministerium für Wirtschaft und
Energie (BMWi), especially M. Gahr (Projektträger FZ Jülich), for funding this scientific work.
Acknowledgments: We gratefully acknowledge the support of the ITVP lab-team. Special thank is also
addressed to M. Tegtmeier (Schaper and Brümmer) for providing the different harvest batches of hawthorn
and fruitful discussions.
Conflicts of Interest: The authors declare no conflict of interest.
Processes 2018, 6, 73 18 of 19

Abbreviations
aP Specific surface area, 1/m
cL Concentration in the liquid phase, kg/m3
cP Concentration in the porous particle, kg/m3
Dax Axial dispersion coefficient, m/s2
Deff Effective diffusion coefficient, m2 /s
KL Equilibrium constant, m3 /kg
Pe Péclet number
q Loading, kg/m3
qmax Maximum Loading, kg/m3
Re Reynolds number
R Radius, m
Sc Schmidt number
SC Side Component
Sh Sherwood number
t Time, s
uz Superficial velocity, m/s
z Coordinate in axial direction, m
ε Dielectric constant, -
ε Voids fraction, -
DPF Distributed plug flow
HPLC High performance liquid chromatography
i.d. Inner diameter
PHWE Pressurized hot water extraction
PTFE Polytetrafluoroethylene
SLE Solid-liquid extraction

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