You are on page 1of 53

The Effect of Processing Parameters on the

Phytochemical Yield of Eurycoma Longifolia


Water Extract Yield

SAIFUL IRWAN ZUBAIRI

PMIFT, Grad B.E.M.

B. Eng. (Chemical-Bioprocess) (Hons.), UTM


M. Eng. (Bioprocess), UTM

ROOM NO.: 2166, CHEMISTRY BUILDING,


TEL. (OFF.): 03-89215828,
FOOD SCIENCE PROGRAMME,
CENTRE OF CHEMICAL SCIENCES AND FOOD TECHNOLOGY,
UKM BANGI, SELANGOR

Outline
Introduction
Tongkat

Ali Background
Tongkat Ali Extraction
Experimental Design
Results and Discussion
Conclusion

Introduction

Global Market growing at 15-20% growth

RM4.55 Billion Malaysian Market

USD 70 Billion market for nutraceuticals


USD 20 Billion market for phytomedicines

80 % imported

Tongkat Ali, Eurycoma Longifolia

Anti-Malarial, Aphrodisiac, Energy Boosting


Malaysia-MIT Biotechnology Partnership Programme
MAVCAP RM20 million invested

Standardisation
Phytomedicine
Standardized extract (100)

Value
added

Freeze/spray dried extracts (40)


Non-standardized extract (25)
Dried powder (10)
Fresh materials (roots, leaves, etc) (2)

Source: Prof Dr. Zhari Ismail, USM

Engineering Questions
What

are critical process parameters?


How do we maximise yield?
What are the economically optimal operating
conditions?
How can we scale up the process?
How do we ensure active ingredient is
present and in the correct amounts i.e.
Standardisation

Processing Technology

Based on traditional method

Food Technology oriented

Need to overcome

Limited concentration in raw material


Solvent cost

More data needed for

Optimisation
Scale up

Objective and Scope


To

develop a mathematical model for the


mass transfer in Batch Solid Liquid Extraction
of Tongkat Ali

Limited

to

Single stage water extracts


Eurycomanone as marker
Optimisation & Scale Up studies

Tongkat Ali background

Biology
Chemistry
Pharmacology
Analysis

Biology and Phytochemistry


Biology

Part of Simaroubaceae family


Slow growing plant, 6-7 years to maturity

Phytochemistry

Quassinoids

Major Component is Eurycomanone

Alkaloids

Highest concentration is 9-Methoxycanthin-6-one

Alkaloids and Quassinoids


OH

HO
OH
O

R
Me

S
S

MeO
O

9-Methoxycanthin-6-one

CH 2

R
O
R

R
R

OH
OH R

R
H

Me

Eurycomanone

Pharmacology
Traditionally

Used for anti-diarrhoea, postpartum tonic, for treating


wounds, boils, and syphilis, anti-pyretic, anti-malarial, antiulcer, energy boosting, and aphrodisiac applications.
Root boiled and decoction drunk

Scientifically

Definite anti-malarial properties (Kardono et al, 1991)


Increases Testosterone production (Farzaturradiah, 1994)
Possibly improves sperm quality (Farzaturradiah, 1994)
Confirmed aphrodisiac effect as Viagra (Pihie, 2003)
Anti tumour properties (Itokawa, 1992)

Analysis
Large

number of compounds >20


Difficulty in identification and quantification
Methods

used include:

Thin Layer Chromatography


UV-Vis Spectrophotometer
High Performance Liquid Chromatography
Liquid Chromatography/Mass Spectrometry

Compounds
Major quassinoids

eurycomanone
longilactone
eurycomalactone
15-acetyl-14hydroxyklaineanone
6-hydroxyeurycomalactone
14,15dihydroxyklaineanone
1,12,15triacetyleurycomanone

Major alkaloids

9,10-dimethoxycanthin-6one
10-hydroxy-9methoxycanthin-6-one
11-hydroxy-10methoxycanthin-6-one
5,9-dimethoxycanthin-6one
9-methoxy-3methylcanthin-5,6-dione

Thin Layer Chromatography

Solvent mixture based on Zhari et al (1999)


Only detects Alkaloids at 365nm
Does not detect quassinoids
Rf=0.86
Light Florescent
Yellow- Green

Rf=0.25
Light Florescent
Green

Rf=1.0
Light Florescent
Blue
Rf=0.69
Light Florescent
Blue

UV Vis Spectrophotometer

Can be calibrated at 238 nm for extract concentration

High Performance Liquid


Chromatography

Based on Chan et al (1998)


First Peak is Eurycomanone, as confirmed by LC/MS

High Performance Liquid


Chromatography
Eurycomanone Calibration
60

y = 3.1087x
R2 = 0.9997

50

ppm

40

30

20

10

0
0

10

Area (x1E6)

12

14

16

18

Tongkat Ali Extraction


Process
Modelling
Optimisation
Scale

Up

Process
Tongkat Ali

Water Extract

Deionized Water

EXTRACTOR
HOLDING
TANK

Hot Air

SPRAY DRYER
Extract
+ fibre
Dehydrated water extract
(final product)

FILTER

Slurry

Modelling
In solid liquid extraction 4 phenomena occur:
1.
2.
3.
4.

The solvent diffuses into the herb particle


The solute is dissolved by the solvent
The solute diffuses to the surface of the herb particle
The solute is dissolved into the bulk solution
1

Cf
4

Cs

3 C*
2

The extraction usually is dominated by 3 or 4

Factors affecting extraction


1.
2.
3.
4.
5.
6.
7.

Solvent or solvent mixture utilised


Solvent to Raw Material Ratio
Raw Material Particle size
Temperature of Extraction
Duration of Extraction
Extraction vessel agitation speed
Extraction vessel volume

Mass Transfer Model


Yield/Concentration

= f (Ratio, Particle Size,


Temperature, Duration, Agitation, Volume)

Can

be done through:-

Theoretical model
Response Surface Methodology
Artificial Neural Networks

Theoretical Model
Liquid Mass Transfer Coefficient, kL

Need to determine relationship between all factors


Mass Balance on vessel

dC f
dt

k L A(C * C f )

Can be rewritten in a exponential form

C f C f
C f C fo

kA
t
V

Agitation increases Mass Transfer Coefficient, kL, to


a maximum value
Diffusion of marker, DAB, in liquid is a critical factor as
well

Theoretical Model
Solid Diffusion, Ds
If

Solid Diffusion, Ds, is the controlling factor,


the mass transfer coefficient, kL, is
determined by it
Based on work by Schwartzberg and Chao
(1982) and Spiro and Selwood (1984)
Estimates of Solid Diffusion for plant material
is around 10-10-10-12 m2/s (Doulia et al, 2000)

Theoretical Model
Other factors
Based

Partition Coefficient, K

on Spiro and Selwood (1984)

Can estimate C*

Weight Fraction of marker/extract, x0

Response Surface Methodology


Statistical-mathematical

method

Design of Experiment
Quantitative Data
Builds model
Optimises

Good

for selecting data


No prior knowledge required of process
May not be able to extrapolate well
Model limited to system studied

Artificial Neural Networks

Model data with unknown structure


Good for complex models (de Villiers & Barnard,
1992)
Can get good results with proper data selection and
treatment (Baratti et al, 1998)
Limited in extrapolation

Optimisation
Method

of choosing best operating point to


maximise desired output i.e. yield or
concentration or profit

Based

on the function obtained from the


Mass Transfer Modelling, it is likely that we
will use

One dimensional constrained optimisation, or


Multivariable constrained optimisation

Scale Up

Produce an identical process result at a larger production rate i.e.


larger extraction vessel

Need defined relationship i.e.

Two key methods

plant

N 3D2

pilot

Basis

P
N 3D2
V

Choose logical basis i.e. P/V


Scale up based on chosen basis

Dimensional analysis (pi matrix/Buckingham method)

Maintain geometrical similarity and identical relevant dimensionless numbers


i.e.

Sherwood and Schmidt for Mass Transfer


Reynolds for fluid flow

Dimensionless Numbers
Reynolds

Number
Dv Da N
Re

Sherwood

Number
kL D
Sh
DAB

Schmidt

Number

Sc
DAB

Experimental Design
Preliminary Experiments (1 litre and 5 litre Glassware)
1. Analytical Method development
2. Determination of relevant parameters
3. Fix value for particle size and agitation rate

Benchscale Experiments (5 litre Pressure vessel)


1. Perform Experiments as per Experimental Design i.e.
General Factorial with 1 replicate
2. Determine optimal point
3. Central Composite Design around Optimal point
4. Calculate Ds, k, NRe and NSc for all experiments
5. Build mass transfer model

Scale Up Experiments (300 litre media tank)


1. Base case on optimal point in Benchscale experiments
2. Scale up agitation to match P/V
3. Test scale up requirement for dimensionless number
similarity i.e. NRe and NSc
4. Build Scale up model if necessary

Preliminary Experiments
Based on UV-Vis calibration to total extract weight

Ratio:

20:1, 30:1, 40:1, 50:1, 60:1 w/w

Duration:

30: 60: 90: 120: 150: 180: 210: 240: 270: 300 min

Particle size:

Smooth (0.5 1.0 mm) and Rough (1 3.5 mm)

Volume:

1 dm3 (Small Scale) and 5 dm3(Large Scale)

Sample:

10g (Small Scale) and 50g (Large Scale)

Total of 10 experiments for each scale with multiple samplings

Preliminary Experiments

Soxhlet extraction apparatus


1 litre small scale

Hydrodistillation apparatus
5 litre small scale

Preliminary Experiments Results


Effect of extraction duration on yield

Yield (%) versus time for same ratio (60:1g/g) at different scale and particle size

Preliminary Experiments Results


Effect of extraction duration on yield

Percent of extraction accomplished with time


(large-scale sample with ratio of 20:1g/g for smooth particles)

Preliminary Experiments Results


Effect of solvent ratio on yield

Yield versus ratio for 1hr sample at different scales and particle sizes

Preliminary Experiments Results


Extraction yield model

Yield versus time for samples with the water to Tongkat Ali ratio of 60:1g/g

Preliminary Experiments Results


Physical Parameters

Density of Tongkat Ali Root

UV Absorbance

Max: approximately 220-240 nm

Extract mass fraction of Tongkat Ali Root

Dry: 0.2g/ml
Wet: 0.6 g/ml

8-10 % w/w

Fraction of Eurycomanone

0.5% of extract w/w

Preliminary Experiments Results


Extraction Parameters

K, partition coefficient

kL, mass transfer coefficient

0.9-1.3 (preliminary)

Small Scale, Smooth particle


Small Scale, Rough particle
Large Scale, Smooth particle
Large Scale, Rough particle

2 x10-6m/s
6 x10-6m/s
2 x10-6m/s
6 x10-6m/s

Ds, Solid diffusion (Schwartzberg & Chao, 1984)

Small Scale, Smooth particle


Small Scale, Rough particle
Large Scale, Smooth particle
Large Scale, Rough particle

9 x 10-12 m2/s
80 x 10-12 m2/s
9 x 10-12 m2/s
80 x 10-12 m2/s

Preliminary Experiments Discussion

Longer duration leads to higher yield

max at 4 to 5 hours
85 % extracted within 30 min and 90% in 1 hr

40:1 ratio best for Smooth and 50:1 ratio best for Rough particles

Higher concentration gradient for mass transfer

Similar yield for Small Scale and Large Scale extractions

Higher yield for smaller particle

More mass transfer area


Lower solid diffusion factor

kL and DS, affected by particle size more than ratio

Need to revise calculation on new data

Preliminary Experiments in
Progress

Other preliminary work in progress


1.
2.
3.
4.
5.
6.

Fine tuning analysis method and apparatus


Calibration of Standard
Effect of agitation
Effect of particle size
Effect of Temperature
Temperature effect of marker degradation

Optimal values of agitation and particle size will be


used for benchscale experiments

Various particle sizes of Tongkat Ali raw material

Benchscale Experiments

Pressurised 5/20 litre heated vessel with agitator

Temperature:
Ratio:
Duration:

80 C, 90 C, 100 C, 110 C, 120 C


20:1, 30:1, 40:1, 50:1, 60:1 w/w
30, 60, 90, 120, 150, 180, 210, 240,
270,300 min

Total of 25 randomised experiments with multiple samplings


2 replicates and repeated analysis of samples
Mass Transfer Model to be built and optimal parameters
determined
Central Composite design around optimal point to confirm
model validity

Expected Benchscale Results

Mass Transfer Model


Surface Response Model
Optimal Operating Point

Properties

Mass transfer coefficient, kL

Solid Diffusion, DS

Reynolds number, Re
Sherwood number, Sh
Schmidt number, Sc

Scale Up Experiments
500

litre Media Tank


Based on Optimal Operating point in
Benchscale experiment
Experiment repeated at larger scale with
central composite design around optimal
point

500 litre Media Tank for Scale up studies

Expected Scale up Results


No

significant difference in yield

Differences

in duration due to heating


process and mixing difference

Scale

up relationship to be formed should


there be a significance difference

Conclusion
Future Work

1.

2.
3.

4.
5.

HPLC Calibration based on analysis from Universiti


Sains Malaysia/FRIM
Determination of optimal particle size and agitation rate
Determination of the effects of temperature on extract
degradation
The benchscale extraction studies
The scale up studies

Conclusion
Recommendations
1.

To acquire chemical standards or independent calibrations as


soon as possible rather than to develop their own standards

2.

To develop a theoretical or empirical model of the process as soon


as possible as well as to take into account variations caused by
organic material

3.

To investigate the effects of multiple stage extraction processes to


reduce utility usage

4.

To perform economic optimisations to determine optimal economic


process parameters

5.

To simulate the process on a batch simulator such as SuperPro


Designer to perform economic evaluation of various design
options

Work Plan
Phase\Month2004

Preliminary

Benchscale

Scaleup

Phase\Month2005

AnalysisofData

Experiments
Writing
DefenceandPublications

10

11

12

10

11

12

X
X

QUESTION AND ANSWER


SESSION

THANK YOU FOR YOUR KIND


ATTENTION

Contribution
Application

of Engineering Methodology
Physical and chemical parameters
Optimal Extraction parameters
General Optimization and Scale up method
for Malaysian Herbs

You might also like