spent coffee grounds as a source of bioactive compounds · spent coffee grounds as a source of...
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3rd Conference ISEKI Food 4Athens, May 2014
Spent Coffee grounds as a source of bioactive compounds
Carmen Monente, Jimena Bravo, Ana I. Vitas, Leire Arbillaga, María Paz de Peña and Concepción Cid
Dept Nutrition, Food Science, and PhysiologyDept. Microbiology and ParasitologyDept. Pharmacology and ToxicologySchool of Pharmacy,University of Navarra (Pamplona, Spain)E-mail: [email protected]
Coffee is one of the main source of antioxidants in diet (Pulido et al., 2003; Pellegrini et al., 2003; Svilaas et al., 2004)
Spent coffee grounds as a source of bioactive compounds
Millions of coffee cups are consumed every day around the World (ICO, 2013)
• Cardiovascular diseases (Bonita et al., 2007)
Coffee may reduce the risk of :
• Cancer (liver, kidney, oral, breast and colorectal) (Nkondjock, 2012)• Neurodegenerative diseases:
Parkinson (Ascherio et al., 2001)
Alzheimer (Barranco Quintana et al., 2007)
Amyotrophic Lateral Sclerosis (Beghi et al., 2011)
• Type 2 diabetes (van Dam & Hu, 2005; Matusheski et al., 2012)
• …
Spent coffee grounds as a source of bioactive compounds
Natural Formed during roasting
Phenolic compounds Maillard Reaction Products (MRPs)Volatiles Non‐volatiles: Melanoidins
Caffeine
• Chemical‐based assays:Scavenger of hydroxyl radicals (Daglia et al., 2004)
• In vitro assays:Prevention of LDL oxidation (Lee et al., 2000) Protection against oxidative DNA damage (Faustmann et al., 2009)
• Chemical‐based assays:Antiradical activity (Nicoli et al., 1997; Daglia et al., 2000;
Del Castillo et al., 2002; Morales & Jimenez‐Perez, 2004)
Metal chelating agents(Borrelli et al., 2002; Delgado‐Andrade et al., 2005)
• In vitro assays:Protection against oxidative stress (Goya et al., 2007) Genoprotective effect (Del Pino‐García et al., 2012)
• In vivo assays:Increase plasma total antioxidant capacity (Somoza et al., 2003)
Protection of LDL oxidation (Dittrich et al., 2009)
• Chemical‐based assays:Antiradical activity (Daglia et al., 2004)
• In vitro assays:Protection against oxidative stress (Pavlica & Gebhardt, 2005; Cho et al., 2009)
• In vivo assays:Protection against oxidative DNA damage (Bichler et al., 2007; Hoelzl et al., 2010)
Chlorogenic acids
Coffee antioxidant compounds
Spent coffee grounds as a source of bioactive compounds
Solubles Non solubles
Coffee compounds extraction
Chlorogenic acidsCaffeineSoluble MelanoidinsHydrophilic volatile compunds
LipidsMelanoidinsPolisacharides
• Extraction method (Bell et al., 2000; Andueza et al., 2003a; Bravo et al. ,2012; Ludwig et al., 2007)
• Grinding (Spiro & Selwod, 1984; Lingle, 1996; Barberá, 2000; Andueza et al., 2003a; Illy & Viani, 2005)
• Coffee/water ratio (Andueza et al., 2007)• Water pressure and Temperature (Andueza et al.,
2002, 2003b, 2007)
Spent coffee grounds as a source of bioactive compounds
Coffee Brew Antioxidant capacity
Per g roasted coffee Per mL coffee brew
> > >
(Pérez‐Martínez et al., 2010; Bravo et al., 2012)
Pérez-Martínez et al., 2010
Spent coffee grounds:Source of natural
antioxidants?J. Agric. Food Chem. 58, 2958-2965
Spent coffee grounds as a source of bioactive compounds
Millions of coffee cups are consumed every day around the World (ICO, 2013)
•Compost•Animal feed (Givens & Barber, 1986)
•Biofuel, biodiesel or bioethanol production (Silva et al., 1998; Kondamudi et al., 2008)
•Adsorbent and activated carbon (Namane et al., 2005; Franca et al., 2009)
•Antioxidants extraction (Yen et al., 2005; Ramalakshmi et al., 2009; Mussatto et al., 2011; Murthy & Madhava Naidu, 2012 )
SOLUBLE COFFEE INDUSTRY
COFFEE SHOPS AND DOMESTIC
•Compost (Cruz et al., 2014)
•Espresso coffee (Zuorro et al., 2013)
Tons of spent coffee grounds
Spent coffee grounds:Source of natural antioxidants?
Spent coffee grounds as a source of bioactive compounds
To explore the possibility to obtain extracts of spent coffee grounds withantioxidant capacity and to evaluate their potential functional properties
1. To develop an extraction methodology of spent coffee with antioxidant capacity.
2. To evaluate the antioxidant capacity and the main bioactive compounds in spentcoffee extracts.
3. To evaluate the potential functional properties of spent coffee extracts:
1. Antioxidant and genoprotective effects in human cells.
2. Antimutagenic activity
3. Antimicrobial activity
OBJECTIVES
Spent coffee grounds as a source of bioactive compounds
1. Extraction methodology (Bravo et al., 2013)
Food Res. Int. 2013, 50, 610-616
Spent coffee groundsSpent coffee:water
1:17
Antioxidant capacityFolin Ciocalteu (Singleton and Rossi, 1965)
ABTS (Re et al., 1999)
DPPH (Brand-Williams et al., 1995)
Browned compounds
(Abs. 420 nm)
Arabica coffee (Guatemala)
Extraction systemSolid-liquid 1:10 (10 min)Filter coffeemaker (6 min)Soxhlet (1h)Soxhlet (2h)
SolventsWater (W) (100%)Water:EtOH (W:E)(80:20,60:40,40:60,20:80)EtOH (E) (100%)Water:MeOH (W:M)(70:30,30:70)MeOH (M) (100%)
Water pH4.57.09.5
Succesive extractions
Defatting
Lyofilization
Spent coffee grounds as a source of bioactive compounds
1. Extraction methodology (Bravo et al., 2013)
0
20
40
60
80
100
120
140
160
1
μmol
Trol
ox/g
dm
Solid-liquid Filter Soxhlet Soxhlet coffeemaker 1h 3h
ab
c d
ABTS Solid-liquid 1:10 (10 min)Filter coffeemaker (6 min)Soxhlet (1h)Soxhlet (2h)
020
406080
100120140
160180
1
μmol
Trol
ox/g
dm
c
e f f e de
d
b
a
W W:Et W:Et W:Et W:Et Et W:Me W:Me Me100 80:20 60:40 40:60 20:80 100 70:30 30:70 100
0
20
40
60
80
100
120
140
160
µmol
Tro
lox
/ g d
m
Extraction Extraction Extraction Extraction Extraction 1 2 3 4 5
020406080
100120140160180200220
1
µmol
Trol
ox/g
dm
pH 4.5 pH 7.0 pH 9.5
c
a
b
Water pH4.57.09.5
SolventsWater (W) (100%)Water:EtOH (W:E)(80:20,60:40,40:60,20:80)EtOH (E) (100%)Water:MeOH (W:M)(70:30,30:70)MeOH (M) (100%)
Food Res. Int. 2013, 50, 610-616
Spent coffee grounds as a source of bioactive compounds
2. Antioxidants in spent coffee grounds extracts (Bravo et al., 2012)
Spent Coffee
Coffee brews Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Robusta coffee(Vietnam)
Arabica coffee(Guatemala)
HPLC: SPE (Bicchi et al., 1995) HPLC method (Farah et al., 2005)
Caffeoylquinic acids: 3-CQA, 4-CQA, 5-CQADicaffeoylquinic acids: 3,4-diCQA, 3,5-diCQA, 4,5-diCQACaffeine
Browned Compounds(Abs. 420 nm)
4-C
QA
325 nm
276 nm
3-C
QA
3,4-
diC
QA
3,5-
diC
QA
4,5-
diC
QA
Caf
fein
e
5-C
QA
Antioxidant capacityFolin Ciocalteau (Singleton and Rossi, 1965)
ABTS (Re et al., 1999)
DPPH (Brand-Williams et al., 1995)
Fremy’s salt & TEMPO(Roesch et al., 2003; Caemmerer and Kroh, 2006)
J. Agric. Food Chem.. 2012, 60, 12565-12573
Spent coffee grounds as a source of bioactive compounds
2. Antioxidants in spent coffee grounds extracts (Bravo et al., 2012)
Spent Coffee
Coffee brews Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Robusta coffee(Vietnam)
Arabica coffee(Guatemala)
Arabica
0
100
200
300
400
500
600
700
Filter Espresso Plunger Mocha Filter Espresso Plunger Mocha
µmol
Tro
lox/
g
a
d
d
b
a b
c
c
spent coffee coffee brew
ABTS
Robusta
0
100
200
300
400
500
600
700
Filter Espresso Plunger Mocha Filter Espresso Plunger Mocha
µmol
Tro
lox/
g
a
d
c b
a b
c
d
spent coffee coffee brew
ABTS
J. Agric. Food Chem.. 2012, 60, 12565-12573
dicaffeoylquinic acid
Spent coffee grounds as a source of bioactive compounds
Spent Coffee
Coffee brews Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Robusta coffee(Vietnam)
Arabica coffee(Guatemala)
caffeoylquinic acid
Arabica
0
5
10
15
20
25
30
Filter Espresso Plunger Mocha Filter Espresso Plunger Mocha
mg/
g
spent coffee coffee brew
nd
CGARobusta
0
5
10
15
20
25
30
Filter Espresso Plunger Mocha Filter Espresso Plunger Mocha
mg/
g
spent coffee coffee brew
nd
CGA
Total CQAs(3-CQA, 4-CQA, 5-CQA)
Total diCQAs(3,4-diCQA, 3,5-diCQA, 4,5-diCQA)
2. Antioxidants in spent coffee grounds extracts (Bravo et al., 2012)
J. Agric. Food Chem.. 2012, 60, 12565-12573
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extracts
Spent coffee
Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Aqueous extraction(Bravo et al., 2013)
Arabica coffee(Guatemala)
Robusta coffee(Vietnam)
Spent coffee
Spent coffee extracts
1. Antioxidant (ROS) and genoprotective (Comet assay) effects in human cells
2. Antimutagenic activity
3. Antimicrobial activity
Food Chem. Toxicol. 2013, 60, 397-403
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extractsAntimutagenic activitySalmonella mutagenicity test (Ames test) S. Typhimurium His−, TA98 strain•Toxicity assay: non‐toxic concentration selection•Antimutagenicity test:
• Without metabolic activation (‐S9): sample+phosphate buffer+culture+mutagen (NPD)• With metabolic activation (+S9): sample+rat liver homogenate (S9mix)+culture+mutagen (NPD)
48h 37ºCRevertant colonies
Robusta EspressoArabica Filter
Spent coffee: Up to 92% (+S9)
Monente et al. Under revision
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extracts
Spent coffee
Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Aqueous extraction(Bravo et al., 2013)
Arabica coffee(Guatemala)
Robusta coffee(Vietnam)
Spent coffee
Spent coffee extracts
1. Antioxidant (ROS) and genoprotective (Comet assay) effects in human cells
2. Antimutagenic activity
3. Antimicrobial activity
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extractsAntimicrobial activity
Spent coffee
Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Aqueous extraction(Bravo et al., 2013)
Arabica coffee(Guatemala)
Robusta coffee(Vietnam)
Spent coffee
Spent coffee extracts
1. Agar-well diffusion method2. Minimum inhibitory concentration (MIC)3. Growth inhibition curves
Gram +Staphylococcus aureusListeria monocytogenesBacillus subtilis
Fungal strainsCandida albicansAspergillus niger
Gram -Escherichia coliPseudomonas aeruginosaSalmonella Choleraesuis
Monente et al. Under revision
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extractsAntimicrobial activity
Inhibition zones (mm)Arabica Filter Robusta espresso
Organisms Spent coffee
Coffee brewSpent coffee
Coffee brew
S. aureus 17.3 ± 0.6ab 15.3 ± 0.6a 15.3 ± 1.2a 19.7 ± 1.2b
L. monocytogenes 20.0 ± 3.6a 19.0 ± 2.7a 18.0 ± 2.0a 22.3 ± 3.1a
B. subtilis 8.8 ± 0.3a 9.7 ± 0.6a ND 11.9 ± 0.2 a
E. coli ND ND ND 10.3 ± 1.2S. Choleraesuis ND ND ND 10.7 ± 0.6Ps. aeruginosa ND ND ND 10.3 ± 0.6C. albicans 15.3 ± 1.2ab 13.7 ± 0.6a 15.7 ± 1.5ab 17.7 ± 0.6 b
A. niger ND ND ND ND
1. Agar‐well diffusion method
Gram +
2. Minimum inhibitory concentration (MIC)MIC (mg/mL)
Arabica filter Robusta espressoOrganisms Spent
coffeeCoffee brew
Spent coffee
Coffee brew
S. aureus 5 7.5 10 8.1L. monocytogenes 20 30 20 16.3B. subtilis 40 60 40 32.5E. coli 80 60 160 32.5S. choleraesuis 40 30 40 16.3Ps. aeruginosa 40 60 80 16.3C. albicans 40 60 40 32.5
Gram +
Monente et al. Under revision
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extractsAntimicrobial activity3. Growth inhibition curves
♦ Control■ Arabica spent coffee▲Robusta spent coffee
Monente et al. Under revision
Spent coffee grounds as a source of bioactive compounds
3. Potential Functional properties of spent coffee extracts
Spent coffee
Spent coffee extracts
Aqueous extraction(Bravo et al., 2013)
Aqueous extraction(Bravo et al., 2013)
Arabica coffee(Guatemala)
Robusta coffee(Vietnam)
Spent coffee
Spent coffee extracts
1. Antioxidant (ROS) and genoprotective (Comet assay) effects in human cells
2. Antimutagenic activity
3. Antimicrobial activity
Spent coffee grounds as a source of bioactive compounds
Spent coffee grounds are a good source of natural bioactive compounds(antioxidants) and it can be considered as a potential preservative to extentfood shelf‐life or to applied to develop functional foods.
CONCLUSIONS
1. To optimize technological extraction conditions, mainly at industry level
2. To study the addition of spent coffee grounds to other foods or productsto develop functional foods (also in terms of sensorial properties) or othernutraceutical, cosmetic or pharmaceutical products
3. to study the bioavailability of bioactive compounds
4. To know antioxidant activity in animal and human studies
But… further research are needed:
Spent coffee grounds as a source of bioactive compounds
ACKNOWLEDGMENTS
• Coffee Research Team• Toxicology lab• Microbiology labUniversity of Navarra
Financed by:• Spanish Minister of Economy and Competitiveness (AGL2009-12052)• Asociation of Friends of the University of Navarra (Grants)• Unión Tostadora S.A.
3rd Conference ISEKI Food 4Athens, May 2014
Spent Coffee grounds as a source of bioactive compounds
Carmen Monente, Jimena Bravo, Ana I. Vitas, Leire Arbillaga, María Paz de Peña and Concepción Cid
Dept Nutrition, Food Science, and PhysiologyDept. Microbiology and ParasitologyDept. Pharmacology and ToxicologySchool of Pharmacy,University of Navarra (Pamplona, Spain)E-mail: [email protected]