spent coffee grounds as a source of bioactive compounds · spent coffee grounds as a source of...

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)

• …


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


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)


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


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?


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


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


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


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





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


µmol

Tro

lox/

g

a

d

c b

a b

c

d


ABTS

J. Agric. Food Chem.. 2012, 60, 12565-12573

dicaffeoylquinic acid


Spent Coffee





caffeoylquinic acid

Arabica

0

5

10

15

20

25

30


mg/

g


nd

CGARobusta

0

5

10

15

20

25

30


mg/

g


nd

CGA

Total CQAs(3-CQA, 4-CQA, 5-CQA)

Total diCQAs(3,4-diCQA, 3,5-diCQA, 4,5-diCQA)


J. Agric. Food Chem.. 2012, 60, 12565-12573


3. Potential Functional properties of spent coffee extracts

Spent coffee

Spent coffee extracts





Spent coffee


1. Antioxidant (ROS) and genoprotective (Comet assay) effects in human cells



Food Chem. Toxicol. 2013, 60, 397-403


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






Spent coffee






3. Potential Functional properties of spent coffee extractsAntimicrobial activity

Spent coffee






Spent coffee


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



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 +



3. Potential Functional properties of spent coffee extractsAntimicrobial activity3. Growth inhibition curves

♦ Control■ Arabica spent coffee▲Robusta spent coffee




Spent coffee






Spent coffee






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:


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]

spent coffee grounds as a source of bioactive compounds · spent coffee grounds as a source of...

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