1. Separation Techniques
Separation of health compounds in Goji (Lyceum barbarum) aqueous extracts by membrane technology
Carmela Conidi, Alfredo Cassano, Enrico Drioli
Institute on Membrane Technology, ITM-CNR,
c/o University of Calabria, via P. Bucci, 17/C
I Rende (CS), Italy
Phone:
Fax:
2nd International Conference and Expo on
Separation Techniques
September , 2016 Valencia, Spain

2. Goji berries and leaves: state of the art and health properties
Outline
Goji berries and leaves: state of the art and health properties
Membrane processes for the treatment of Goji berries and leaves aqueous extracts
Clarification of Goji aqueous extracts
Fractionation of clarified extracts
Analysis of membrane productivity
Analysis of membrane selectivity (antioxidant activity, total polyphenols, soluble solids, total carbohydrates)
Conclusions

3. State of art
Goji berries are one of the top 10 “Super Food” cultivated in the world
Popular in traditional Chinese medicine as well as Western parts of the world
The plant is also cultivated in the Mediterranean region (goji berries grow very well in regions with semi-arid to sub-tropical climatic conditions)

4. Global market of gojy extracts

5. Health benefits
H. Amagase , N. R. Farnsworth. A review of botanical characteristics, phytochemistry, clinical relevance in efficacy and safety of Lycium barbarum fruit (Goji), Food Research International 44 (2011) 1702–1717

6. Bioactive compounds b-carotene Polyphenols Ascorbic acid
Polysaccarides
D. Donno, G.L. Beccaro, M.G. Mellano, A.K. Cerutti, G. Bounous. Goji berry fruit (Lycium spp.): antioxidant compound fingerprint and bioactivity evaluation, Journal Functional Foods, 2015,

7. ADVANTAGES LIMITATIONS
Recovery of bioactive compounds from vegetable sources
Ultrasound assisted extraction
Solvent extraction
Resin-based extraction
Supercritical fluid extraction
Ultrafiltration
Nanofiltration
Reverse Osmosis
ADVANTAGES
LIMITATIONS
more expensive than aqueous processing
residual solvents in the extracts (no desiderable for food use)
easy denaturation of thermo-sensitive compounds
reduction of nutritional value
low energy requirement
additive free (do not require the use of organic solvents)
mild operating conditions
preservation of thermo-sensitive compounds

8. Clarification/Fractionation with membranes processes
Feed
Permeate
Retentate
Membrane
Driving force = DP
Pressure-driven membrane processes
Process
Membrane
DP range
Species retained MF
Porous
0.1 – 5 mm
0.1 – 3 bar
Cells, bacteria, yeasts, starch granules, oil globules, etc. UF
5 – 100 nm
1 – 10 bar
Polysaccharides, proteins, tannins, virus, etc. NF
1 – 5 nm
10 – 50 bar
Sugars, organic acids, polyphenols, aroma compounds, etc. RO
Non porous
10 – 100 bar
Salts UF

9. Goji berries extracts / flow sheet of the investigated process
Aqueous extract
Sieving
(200 mm)
Washing
Milling
CLARIFICATION
(UF)
Retentate
b-carotene
Clarified extract
Extraction with water
(200 g/kg, 80 °C, 30 min)
FRACTIONATION
(UF/NF kDa)
Separation of carbohydrates from polyphenols

10. China Blue Star Membrane Technology Co. Ltd.
Clarification of goji berries extracts
Manufacturer
Toray
China Blue Star Membrane Technology Co. Ltd.
Membrane type
HFS
DCQ II-006 C-PS100
Configuration
Hollow fiber
Membrane material
Polyvinylidene fluoride
Polysulphone
Max oper. pressure (bar) 2
1.5
Max oper. temperature (°C) 40 50
pH range
2-12
2-13
Membrane surf. area (m2)
0.4
0.16
Dimension of pores
0.02 mm -
MWCO (kDa)
100
Toray HFS
UF PS DCQ

11. (Operating conditions: TMP, 0.7 bar; Qf, 280 L/h; T, 24±2°C)
Clarification of goji berries extracts
FEED
Permeate
Retentate
Feed
Permeate
Retentate
Ultrafiltration of goji berries aqueous extracts. Permeate flux vs. VRF
(Operating conditions: TMP, 0.7 bar; Qf, 280 L/h; T, 24±2°C)

12. Clarification of goji berries extracts
PS membrane
Parameters
FEED
PERMEATE
RETENTATE
Total suspended solids (%)
3.2±1.1 -
3.8±0.2
Total soluble solids (°Brix)
4.5±0.4
4.0±0.2
4.5±1.2
b-carotene (mg/ml)
2.80±0.5
0.260±0.05
4.74 ±0.25
Total polyphenols (mg/L gallic acid)
520.5±2.6
510.0±6.0
540.0±8.0
TAA (mM Trolox)
3.5±0.5
2.6±0.2
3.65±0.12
Total carbohydrates (g/L glucose)
15.1±2.3
14.35±1.52
18.2±2.3

13. Clarification of goji berries extracts
PVDF membrane
Parameters
FEED
PERMEATE
RETENTATE
Total suspended solids (%)
3.1±0.2 -
3.8±0.4
Total soluble solids (°Brix)
4.2±0.1
4.1±0.2
b-carotene (mg/ml)
3.16±0.12
0.098±0.02
4.34±0.35
Total polyphenols (mg/L gallic acid)
448.5±2.6
400±6.0
480±6.0
TAA (mM Trolox)
2.8±0.3
2.15±0.25
2.6±0.4
Total carbohydrates (g/L glucose)
17.7±3.2
13.5±2.6
18.67 ±2.4

14. Retention of UF membranes towards analyzed compounds
Clarification of goji berries extracts
Retention= (1-Cp/Cf)*100
b-carotene
TAA
TSS
Total polyphenols
Total carbohydrates
Total suspended solids
Retention of UF membranes towards analyzed compounds

15. Clarified goji berries aqueous extracts
Fractionation of clarified goji berries extracts
Clarified goji berries aqueous extracts
Membrane type
Desal GE
Desal GH
Desal GK
Manufacturer
GE Water & Process
Membrane material
thin film composite with a cross-linked
aromatic polyamide top layer
Molecular weight cut-off (Da)
500
1000
2000
Max. operating pressure (bar) 27
Max. operating temperature (°C) 90
Range of pH
1-11

16. (Operating conditions: Qf, 300, L/h; T, 24±2°C)
Fractionation of clarified goji berries extracts
Treatment of clarified Goji aqueous extracts with UF and NF membranes. Evolution of permeate flux as function of TMP
(Operating conditions: Qf, 300, L/h; T, 24±2°C)

17. Fractionation of clarified goji berries extracts
Retention of tested membrane towards total polyphenols
Retention of tested membrane towards TAA
Retention of tested membrane towards TSS
Retention of tested membrane towards total carbohydrates

18. Fractionation of clarified goji berries extracts
Retention of GK membrane towards analyzed compounds
Retention of GH membrane towards analyzed compounds
Retention of GE membrane towards analyzed compounds

19. (Operating conditions: TMP, 10 bar; T, 27±1°C)
Diafiltration/concentration
diafiltrate
Water UF
permeate
Desal GE
Membrane
Time evolution of permeate flux during diafiltration/concentration processes
(Operating conditions: TMP, 10 bar; T, 27±1°C)

20. Diafiltration/concentration
Analyses of TSS, total polyphenols, total carboydrates and TAA in samples of gojy berries extracts coming from the diafiltration/concentration processes
Diananofiltration*
Concentration in batch**
Parameters
Initial
Final
Total soluble solids (°Brix)
4.5±0.4
0.4±0.02
0.8±1.2
Total polyphenols (mg/L gallic acid)
520.5±2.6
482.4±8.2
812.4±10.2
TAA (mM Trolox)
3.5±0.5
2.9±0.6
4.8±0.9
Total carboydrates (g/L glucose)
15.1±2.3
6.2±0.2
7.4±0.5
*After 4.5 volumes of diafitration
** Weight reduction factor (WRF) of 2

21. extraction parameters (T, pH, solid/liquid ratio)
Goji leaves / Flow sheet of the investigated process
Goji leaves
Aqueous leaf extract
Washing
CLARIFICATION
(UF 100 kDa)
Clarified extract
Retentate
drying
FRACTIONATION
Extraction with water
UP004
NP010
N30F
Optimization
extraction parameters
(T, pH, solid/liquid ratio)
(Mycrodin Nadir)
flat-sheet
Polyethersulphone
MWCO = 4,000 Da
(Mycrodin Nadir)
flat-sheet
Polyethersulphone
MWCO = 1,000 Da
(Mycrodin Nadir)
flat-sheet
Polyethersulphone
MWCO = Da

22. Optimization of extraction parameters
Effect of concentration (solid/liquid ratio) on the TSS, Total polyphenols and TAA yields

23. HPLC cromatograms of polyphenols in goji leaves extracts
Optimization of extraction parameters
min 5 10 15 20 25 30 35 40
mAU
100
200
300
400
500
600
700
800 .
Ethanol/water (1:1) extraction: concentration 10 g/Kg
Aqueous extraction: concentration 10 g/Kg
Aqueous extraction: concentration 100 g/Kg
HPLC cromatograms of polyphenols in goji leaves extracts

24. Optimization of extraction parameters
Effect of temperature of the extraction on the TSS and Total polyphenols yields

25. Optimization of extraction parameters
Effect of the pH of the extraction solution on the TSS and Total polyphenols yields

26. Clarification of goji leaves extracts
DCQ II-006 C-PS100
Hollow fiber
Polysulphone
100 kDa
Ultrafiltration of goji leaves extracts.
Time course of permeate flux and VRF
(Operating conditions: TMP, 0.7 bar; Qf, 280 L/h;
T, 24±2°C)
Parameters
Feed UF
Permeate UF
Retentate UF pH
7.2±0.1
7.1±0.2
7.2±0.8
Total suspended solids
5.2±2.3 -
6.2±0.1
Total soluble solids (°Brix)
10.5 ±0.2
9.5±0.12
12±1.2
Total polyphenols (mg/L gallic acid)
±4.5
±2.8
2570 ±2.6
TAA (mM Trolox)
13±1.2
11±2.6
14±1.6
Total carbohydrates (g/L glucose)
6.33±1.6
5.43±0.12
9.65±2.8

27. (Operating conditions: Qf, 300 L/h; T, 24±2°C)
Fractionation of clarified goji leaves extracts
Treatment of clarified leaves aqueous extracts with UF and NF membranes . Evolution of permeate flux as function of TMP
(Operating conditions: Qf, 300 L/h; T, 24±2°C)

28. Fractionation of clarified goji leaves extracts
Retention of NP010 membrane towards analyzed compounds
Retention of UP004 membrane towards analyzed compounds
Retention of N30F membrane towards analyzed compounds

29. Total polyphenols (mg/L gallic acid) Total carbohydrates (g/L glucose)
Fractionation of clarified goji leaves extracts
Analyses of total polyphenols, TAA, total soluble solids and total carbohydrates in
goji leaves samples from NPO10 membrane at different WRF
Type of sample
Total polyphenols (mg/L gallic acid)
TAA
(mM Trolox)
Total
soluble solids
(%)
Total carbohydrates (g/L glucose)
Feed
1520 ± 1.52
9.54 ± 1.47
10.2 ±0.2
5.61 ±0.01
Permeate WRF 1,5
± 0.85
2.47 ± 0.23
5.6±0.05
4.84 ± 0.02
Permeate WRF 2
± 0.20
3.02 ± 0.78
6.2±0.4
4.63 ± 0.05
Final permeate
± 0.61
2.80±0.1
6.4±0.1
4.85 ± 0.4
Retentate WRF 1,5
± 1.17
10.49 ± 2.26
6.5±0.3
5.93 ± 0.8
Retentate WRF 2
± 0.42
15.32 ± 0.99
6.8±0.25
5.56 ± 0.2
Retention of NPO10 membrane towards analyzed compounds

30. Conclusions
A combination of aqueous extraction and membrane-based purification processes of bioactive compounds from Goji berries and leaves has been studied and optimised.
The PVDF UF membrane allows to separate b-carotene in Goji berries aqueous extracts with high efficiency in terms of retention (98%) and productivity (more than 40 kg/m2h).
Operating conditions (pH, temperature, concentration) for the aqueous extraction of goji leaves have been optimized.
In the fractionation of both clarified extracts permeate fluxes increased linearly with the operating pressure. At low pressure values a better separation of antioxidant compounds (including polyphenols) from carbohydrates and TSS has been observed.
The sugar purification can be improved significantly if diafiltration is applied.
A strict correlation between the MWCO of selected membranes and the retention of investigated compounds has been detected.

31. Tank you…for your kind attention!