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Oleoresins from Capsicum spp.: Extraction

Methods and Bioactivity

E. AzuÌara-Nieto, Guiomar Melgar, Alan Javier Hernández Alvarez

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ORIGINAL PAPER

Oleoresins from Capsicum spp.: ExtractionMethods and Bioactivity

Guiomar Melgar-Lalanne1 & Alan Javier Hernández-Álvarez2 &

Maribel Jiménez-Fernández1 & Ebner Azuara1

Received: 7 January 2016 /Accepted: 16 August 2016 /Published online: 9 September 2016# Springer Science+Business Media New York 2016

Abstract Capsicum spp. fruit is one of the most producedvegetables around the world, and it is consumed both as freshvegetable and as a spice like a food additive for their character-istic red color and, in many cases, its pungency. In addition toits economic importance, the bioactivity of some importantcompounds such as capsaicinoids and carotenoids has promot-ed its research. The use of Capsicum oleoresins has been in-creased due to its advantages comparing with the traditional dryspice. These include obtaining higher quality products with thedesired content of bioactive and flavored substances. The widediversity of extraction methods including water extraction, or-ganic solvent extraction, microwave-assisted extraction, andultrasound assisted extraction as well as supercritical fluid ex-traction among others are discussed in the present review.Moreover, pretreatments such as chemical treatments, osmoticdehydration, sun and oven drying, and freeze-drying common-ly used before the extraction are also presented. Due to itsimportance, Capsicum oleoresins produced with Bgreen^ sol-vents and the improvement of fractional extraction techniquesthat allow to obtain separately the various bioactive fractionswill continue under research for further development.

Keywords Capsicum spp. extraction . Oleoresin . Essentialoil . Bioactivity . Antioxidant activity

Introduction

TheCapsicum genus consists, up to date, of 31 species, five ofwhich are used as fresh vegetables and species: Capsicumannum, Capsicum baccatum, Capsicum chinense, Capsicum

frutescens, and Capsicum pubescens (Moscone et al. 2007).The center of domestication and dispersal patterns of thesespecies remains speculative, although it has been suggestedthat C. annum was initially domesticated in Mexico,C. frutescens in the Caribbean, C. baccatum in Bolivia, andC. pubescens in south of the Andes (Perry et al. 2007). Ingeneral, Capsicum spp. are commonly divided/distributed/split into two main groups, depending on their Scoville heatunits (SHU), a measurement of their pungency: sweet or non-pungent fruits (syn. bell pepper, pepper, paprika, moron chili,or sweet chili) and hot or pungent fruits (syn. chili, red pepper,hot pepper, spicy pepper, cayenne, hot paprika). Only inMexico, there are about a hundred cultivars of hot Capsicumspp., more or less spicy, more or less large, with a long orrounded shape and colors varying from pale yellow to darkred. Some varieties of C. annuum L. (Serrano, Jalapeño,Poblano, Guajillo) and C. chinense (Habanero) are currentlycultivated worldwide (Katz 2009). Their particular character-istics of pungency, aroma, and flavor made this product animportant ingredient in millions of people’s daily diets(Meckelmann et al. 2015). In many Latin American andAsian countries, it is an essential part of their daily cooking;in India, it is a basic ingredient in the traditional curry blends,and in the Mediterranean region, it is widely used as dye,preservative, and seasoning in many meat sausages both freshand dehydrated. Green and red bell peppers at different matu-rity stages can be from the same cultivar. Green fruits areimmature and usually consumed as fresh or minimally proc-essed vegetables and red fruits are consumed both as freshvegetables and as spice (as powder). Nowadays, red

* Ebner Azuaraeazuara@uv.mx

1 Instituto de Ciencias Básicas, Universidad Veracruzana, Av. Dr. LuisCastelazo Ayala s/n. Col Industrial Ánimas, 91192 Xalapa, Veracruz,Mexico

2 Food Research and Development Center, Agriculture and Agri-FoodCanada, 3600 Casavant West, St. Hyacinthe, QC J2S 8E3, Canada

Food Bioprocess Technol (2017) 10:51–76DOI 10.1007/s11947-016-1793-z

dehydrated fruits in powder or as oleoresins are commonlyused to modify color and flavor of many dishes such as soups,stews, sausages, cheese, snacks, salad dressings, pizza, con-fectionaries, and beverages (Arimboor et al. 2015). Moreover,dehydrated chili powder is used in poultry feed as egg pigmentand as prophylactic antimicrobial against some pathogens(Vicente et al. 2007; Lokaewmanee et al. 2013).

Fruits of Capsicum spp. vary in color, sharp, and size be-tween and within species. Ripe fruits display a range of colorsfromwhite to deep red. The intensity of red color and the degreeof pungency are valued as major quality parameters. The pun-gency depends on capsaicinoids content which in turns dependson the variety and maturation stage and is commonly measuredin Scoville heat units (SHU). The red color is imparted mainlyby carotenoids with more than 50 identified structures.Capsainthin, capsorubin, and cryptocapsin impart brilliant redcolor to ripen fruits while the yellow orange color is given byβ-carotene, zeaxanthin, violaxanthina, and b-cryptoxantin(Hornero-Méndez and Mínguez-Mosquera 2001; Cervantes-Paz et al. 2012). Capsicum spp. fruits are considered as non-climacteric fruits. Green or deep green harvest fruits have failedto reach a fully red color after harvest while fruits that wereharvested at or after the breaker stage completed their colorchange to a fully red. However, the different color stages inmature fruits affect the color development and quality, but notthe pungency level. Therefore, to ensure the highest quality, it isnecessary to harvest the fruit when a completely red color hasbeen developed in the plant (Krajayklang et al. 2000).

Despite being one of the most consumed vegetables world-wide, there is scarce information about the proximate compo-sition of different commercial Capsicum spp. both in dried orfresh forms (see Table 1) and, in general, is incomplete. OnlyEsayas et al. (2011) determined fiber as crude fiber andcarbohydrates as available carbohydrates, and Abdou Boubaet al. (2012) also determined the available carbohydrates, butnot crude fiber hinders the comparison with the rest; most ofauthors determined the total carbohydrates. Most of the arti-cles do not specify the botanical species of the analyzed chilior the maturity grade. In commercial dried Capsicum spp.,moisture varies from 5.5 to 13.0 g/100 g of fresh sample whilein fresh Capsicum spp. the moisture varies from 89.90 to94.1 g/ 100 g fresh sample, depending the variety analyzed.Orellana-Escobedo et al. (2013) found the highest differencesin carbohydrate (between 69.9 and 58 g/100 g), similar toEsayas et al. (2011) who found similar quantities ofcarbohydrates and crude fiber in common dried Capsicum

spp. Finally, only Abdou Bouba et al. (2012) found minorquantities of carbohydrates in bird Capsicum (7.0 g/100 g)calculated as available carbohydrates without the incorpora-tion of the fiber fraction, but without consider it. Protein is theless variable parameter both in dried and fresh samples, andthe fat portion ranges between 4.2 and 13.8 g/100 g in driedsamples and from 0.16 to 0.54 g/100 g in fresh samples. It is

important to highlight that the maturity grade affected theproximate chemical composition, color, total capsaicinoids,and ascorbic acid contents in analyzed samples (Zaki et al.2013; Martínez et al. 2007).

There are several groups of valuable compounds inCapsicum spp. including carbohydrates, which constitute ap-proximately 85 % of the dry weight, polyphenols (0.5 % of dryweight) and minor but important compounds such ascapsaicinoids, carotenoids, and vitamins (Arimboor et al.2015). The physico-chemical parameters and mineral composi-tion depend directly on the maturity stage of the fruit. Thesoluble solid content and titratable acidity increase during rip-ening as well as the fat, the protein, and the ascorbic acid con-tent (Martínez et al. 2007). The flavor profile of Capsicum spp.from several parts of the world has been reported in manystudies describing the changes in volatile organic compoundsduring ripening, showing that the producer aroma compoundsdiffer significantly during the maturity process (Liu et al. 2009).

In the food industry, the presence of pungent principles(capsaicinoids) in oleoresins of hot Capsicum spp. may repre-sent a limitation for its application as a food dye and restrictsthe exploration of a large number of high yielding pungentvarieties. Efforts to improve extraction methods to preparenon-pungent oleoresins from pungent Capsicum fruits by theselective removal of capsaicinoids are in continuous researchmostly focused in the development of fractionated extractiontechniques (Arimboor et al. 2015).

Drying as Pretreatment for Extraction

Chemical Pretreatments

The use of chemical pretreatments such as sodium and potas-sium hydroxide, potassium meta bisulphate, potassium car-bonate, methyl and ethyl ester emulsions, and ascorbic andcitric acid have been suggested to obtain better quality char-acteristics of dehydrated Capsicum spp. (see Table 2).

Therefore, the treatment with ethyl oleate and citric acidsolutions before the drying process can reduce the drying timeand the mass-transfer resistance both in red pepper (Doymazand Kocayigit 2012) and green pepper (Doymaz and Ismail2013). Red C. annuum L. previously treated with a solutioncontaining NaCl, CaCl2, and Na2S2O3 prior to drying at 70 °Cshowed the best firmness and color quality (Vega-Gálvez et al.2008). The use of a pretreating solution with 2 % of ethyloleate and 5 % of K2CO3 was found to be effective to providethe best yield and color quality in Capsicum spp. dried at50 °C. Moreover, pretreated fruits dried faster and were foundto have the highest drying rate. Color analysis showed that redcolor is preserved better in pretreated fruits and in slices(Doymaz and Pala 2002). Similar results were obtained inoven-drying previously treated with ethyl oleate and citric

52 Food Bioprocess Technol (2017) 10:51–76

acid solutions where the drying, rehydration, and color char-acteristics were significantly influenced by air temperatureand pretreatments (Doymaz and Kocayigit 2012). The colorof paprika is drastically affected by the drying process of redpeppers through non-enzymatic reactions that gives the pow-der a brown tonality. To solve this, the dip in a solution of 2 %of ethyl oleate solutions plus 2 % of NaOH and 4 % of K2CO3

and an air drying temperature of 60 °C is a good solution thatretains the red color of the original fruit (Ergüneş and Tarhan2006). Sodium metabisulfite at a drying temperature of 70 °Ccan provide a more bright red color because the sulfite inhibitsthe non-enzymatic browning reactions. Moreover, soaking theCapsicum in sodium metabisulfite combined with CaCl2 pro-duced the highest color stability (Wiriya et al. 2009).

Blanching inactivates deteriorative enzymes such as perox-idase isoenzymes and reduces the microbiological rate andimproves the color characteristic of the fruits (Schweiggertet al. 2006). In non-pungent C. annuum L. shreds in boiling

water (3 min) followed by pre-treatments with 0.20 % of po-tassiummetabisulfite and 0.50 % of citric acid for 5 min beforedrying in a solar poly tunnel produced a dehydrated productwith high color quality and stability cid (Sharma et al. 2015).Blanching of sweet C. annuum L. at 95 °C for 5 min improvedthe drying rate by reducing the process time (Vengaiah andPandey 2007). In pungent Capsicum spp., blanching slightlyreduces the initial capsaicinoids content but avoids their oxi-dation during storage (Schweiggert et al. 2006).

Osmotic Dehydration

Osmotic dehydration is a process for the partial removal ofwater in which cellular materials are placed in a concentratedsolution of soluble solute through semi-permeable membraneresults in the equilibrium conditions in both sides of mem-brane (Arvanitoyannis et al. 2012). It has been used for im-proving the quality of fruit products and reducing energy

Table 1 Proximate chemical composition (g / 100 g) of different commercial Capsicum spp.

Common name Fresh/dried Moisture Ash Protein Fat Fibera Carbohydrateb Reference

Ancho D 10.1 ± 0.1 7.8 ± 0.0 12.0 ± 0.1 9.8 ± 0.1 nd 60.2 ± 2.3 Orellana-Escobedo et al. 2013

Bako D 9.0 ± 0.2 7.3 ± 0.1 8.7 ± 0.4 9.5 ± 0.1 26.0 ± 1.3 39.5 ± 0.9c Esayas et al. 2011

Bird D 9.3 ± 0.1 9.4 ± 0.2 9.4 ± 0.8 11.1 ± 0.2 nd 60.8 ± 0.1 Abdou Bouba et al. 2012

Chipotle D 8.8 ± 0.2 6.9 ± 0.2 12.7 ± 0.0 8.6 ± 0.8 nd 63.0 ± 1.2 Orellana-Escobedo et al. 2013

Chipotle meco D 8.5 ± 0.4 9.5 ± 0.2 15.2 ± 0.5 9.1 ± 0.0 nd 57.7 ± 1.1 Orellana-Escobedo et al. 2013

De arbol D 5.6 ± 0.2 8.8 ± 0.6 12.7 ± 0.6 13.4 ± 1.0 nd 59.4 ± 0.4 Orellana-Escobedo et al. 2013

Guajillo D 9.1 ± 0.6 7.5 ± 0.0 12.8 ± 0.1 12.4 ± 0.1 nd 58.0 ± 0.6 Orellana-Escobedo et al. 2013

Habanero D 13.0 ± 2.0 7.5 ± 0.0 13.5 ± 0.4 4.6 ± 0.2 nd 61.3 ± 2.4 Orellana-Escobedo et al. 2013

Jalapeño D 10.1 ± 0.9 7.3 ± 0.1 14.3 ± 0.4 4.2 ± 0.2 nd 64.0 ± 0.8 Orellana-Escobedo et al. 2013

Marako fana D 9.2 ± 0.1 5.3 ± 0.6 11.8 ± 0.1 11.2 ± 0.2 27.3 ± 0.2 35.3 ± 0.6c Esayas et al. 2011

Mirasol D 9.9 ± 0.2 9.6 ± 0.1 14.1 ± 0.4 7.5 ± 0.1 nd 59.0 ± 0.5 Orellana-Escobedo et al. 2013

Morita D 10.7 ± 0.9 8.6 ± 1.4 14.1 ± 0.6 7.6 ± 0.2 nd 59.0 ± 1.3 Orellana-Escobedo et al. 2013

Oda haro D 8.8 ± 0.1 7.3 ± 2.2 9.2 ± 0.2 9.2 ± 0.4 28.6 ± 0.8 37.1 ± 2.1c Esayas et al. 2011

Pasado D 8.9 ± 0.5 7.2 ± 1.2 12.6 ± 0.3 5.4 ± 0.1 nd 66.2 ± 1.0 Orellana-Escobedo et al. 2013

Pasilla D 7.6 ± 0.2 5.8 ± 0.0 12.3 ± 0.9 13.8 ± 0.3 nd 60.5 ± 0.8 Orellana-Escobedo et al. 2013

Pepper D 5.70 ± 0.1 4.35 ± 0.1 11.7 ± 0.1 12.7 ± 0.1 2.6 ± 0.0 62.9 ± 0.0c Otunola et al. 2010

Piquin D 5.5 ± 0.1 7.3 ± 0.0 13.7 ± 0.3 11.0 ± 1.0 nd 62.5 ± 1.1 Orellana-Escobedo et al. 2013

Puya D 7.0 ± 0.2 7.8 ± 0.1 13.2 ± 0.3 8.1 ± 0.7 nd 63.8 ± 1.3 Orellana-Escobedo et al. 2013

Serrano D 11.2 ± 0.4 5.8 ± 0.0 12.8 ± 0.6 2.3 ± 0.1 nd 67.9 ± 0.8 Orellana-Escobedo et al. 2013

Tres venas D 9.0 ± 0.0 7.0 ± 0.2 13.3 ± 0.6 9.6 ± 0.1 nd 61.0 ± 0.3 Orellana-Escobedo et al. 2013

Arnoia (green ) F 92.7 ± 1.4 0.40 ± 0.08 0.80 ± 0.15 0.22 ± 0.04 1.63 ± 0.24 3.84 ± 0.78 Martínez et al. 2007

Arnoia (green B) F 93.7 ± 0.1 0.33 ± 0.02 0.71 ± 0.07 0.16 ± 0.03 1.31 ± 0.15 3.51 ± 0.18 Martínez et al. 2007

Arnoia (red) F 89.90 ± 1.2 0.62 ± 0.09 1.13 ± 0.21 0.54 ± 0.15 1.62 ± 0.15 6.23 ± 1.36 Martínez et al. 2007

Green bell pepper F 92.56 0.3 1.05 0.38 0.73 4.98 Faustino et al. 2007

nd not determinedaWhen determined calculated as crude fiberbDetermined as total carbohydrates at least otherwise statedcAvailable carbohydrates

Food Bioprocess Technol (2017) 10:51–76 53

consumption. It is used to reduce the water content of the foodfrom 30 to 70% as an upstream step of the dehydration of foodbefore they are subjected to further processing such as freezing,freezing-drying, vacuum drying, and air drying (Levent andFerit 2014). Since osmotic dehydration as a pretreatment ofmany processes improves nutritional, sensorial, and functionalproperties of food without changing its integrity, it wasexploited as a pretreatment to Capsicum spp. dehydration.During osmotic treatment, mass transfer occurs throughsemi-permeable cell membranes present in biological mate-rials, which offers the dominant resistance to the process.The state of the cell membrane can change from being partiallyto totally permeable, and this can lead to significant changes intissue architecture. Various osmotic agents such as sucrose,glucose, fructose, corn syrup, sodium chloride, and their com-bination have been used for osmotic dehydration (see Table 3).

The addition of small quantities of sodium chloride to os-motic solutions increased the driving force of the drying processand synergistic effects between sucrose and sodium chloridehave been reported (Ade-Omowaye et al. 2002). In general,osmotic dehydration pretreatment has a positive effect in thecolor and other quality attributes of the final dehydratedCapsicum both in the case of red (Falade and Oyedele 2010)and green fruits (Quintero-Chávez et al. 2012). In redCapsicum, a pretreatment with sucrose solutions from 40 to60° Brix resulted in a significant effect on the conservation ofredness value (a*), a mark of color quality of dry red Capsicum(Falade and Oyedele 2010). The osmotic dehydration in green-yellow Padron pepper (C. annuum L. Longum) could be con-sidered as an impregnation process because the solid gain, the

weight reduction, and the water loss increased with the temper-ature and salt concentration. However, the concentration ofNaCl did not develop significant changes in color working atlowest temperatures (25 °C) (Chenlo et al. 2006). The greatlimitation of osmotic dehydration is that the final product showsan intermediate humidity and the product cannot be consideredshelf stable (Levent and Ferit 2014). Most studies related withthe color stability have been done in red fruits. However, ingreen fruits, an important conservation of color was also found(Chenlo et al. 2006) using a combination salt–sorbitol (Ozenet al. 2002; Ozdemir et al. 2008). Salt–sorbitol combinationsignificantly increases weight loss, solids gain, and tissue brixand decreased water activity in green fruits. Moreover, the pres-ence of sorbitol hindered the entrance of salt into the productimproving the sensorial properties of the final product (Ozenet al. 2002; Ozdemir et al. 2008). Temperature is another im-portant parameter to take account because high temperaturescan affect the structure of the fruit in a similar way thatblanching and room temperatures are preferred.

Red C. annuum (varieties Sombo, Rodo, Bawa and Tatase)osmotically dehydrated with sucrose or a combination of su-crose–salt was tested, resulting to higher sucrose concentrationsthat gave better results while improved solute gain were obtain-ed using binary mixture with lower processing time, energy,and cost. Moreover, this pretreatment protects the fruits againstthe discoloration in the posterior sun drying process (RajiAbdul Ganiy et al. 2010). The salt (NaCl) also has differenteffects on osmotic dehydration, depending of the salt source,concentration, temperature, and time. Best results have beenfound with saturated sea salt at 25 °C (Levent and Ferit 2014).

Table 2 Chemical pretreatments used before drying Capsicum spp.

Species Common name Treatment conditions Chemical pretreatments Reference

C. annuum Kahramanmaras Room temperature / 1 min a) 2 % ethyl oleate +4 % K2CO3 Doymaz and Pala 2002b) 2 % ethyl oleate +5 % K2CO3

a

c) 2 % ethyl oleate +6 % K2CO3

C. annuum L. Charliston Room temperature / 1 min a) 2 % ethyl oleate +3 % K2CO3a Doymaz and Kocayigit 2012

b) 0.5 % Citric acid

C. annuum L Green bell pepper Room temperature / 1 min a) 2 % ethyl oleate +3 % K2CO3a Doymaz and Ismail 2013

b) 0.5 % Citric acid

C. annuum L. Paprika 23 °C / 1 min a) 2 % Ethyl oleate (33 °C) Ergünes and Tarhan 200660 °C / 1 min b) 2 % ethyl oleate +2 % NaOH (23 °C)

c) 2 % ethyl oleate +2 % NaOH +4 % K2CO3 (23 °C)

d) 2 % Ethyl oleate (60 °C)

e) 2 % ethyl oleate +2 % NaOH (60 °C)

f) 2 % ethyl oleate +2 % NaOH +4 % K2CO3 (60 °C) a

C. annuum L Lamuyo variety 25 °C /10 min 20 % NaCl +1.0 % CaCl2 + 0.3 % NaS205 Vega-Gálvez et al. 2008

C. annuum L Huarou Yin 90 °C / 3 min a) 0.3 % NaS205 Wiriya et al. 2009b) 0.1 % ascorbic acid

c) 0.3 % NaS205 + 1 % citric acid

d) 0.3 % NaS205 + 1 % CaCl2a

aBest attributes found

54 Food Bioprocess Technol (2017) 10:51–76

In the case of red Capsicum spp. in addition to the combi-nation sorbitol–salt (Quintero-Chávez et al. 2012), the mostwidely used is the combination sucrose–salt (Ade-Omowayeet al. 2002; Zhao et al. 2013). In general, the use of combinedsolutes results in better sensory quality attributes ofdehydrated Capsicum spp. than the solutes alone (Ade-Omowaye et al. 2002). The presence of high levels of sucrosereduces the saltiness and the presence of salt enhances thesucrose sweetness (Sacchetti et al. 2001). Osmotically treatedCapsicum spp. in a mixed solution (10 % NaCl +50 % su-crose) had the best dehydration effect reaching 66 % in waterloss and preventing the penetration of salt to some extentbecause of the existence of sugar showing the preferable colorboth in traditional drying and in microwave drying. However,the samples pretreated with osmotic dehydration and dried bymicrowaves at 60 W needed less drying time (Zhao et al.2013). Similar results were obtained in other research whereCapsicum was osmotically pretreated previously to be driedby microwaves concluding that the pretreatment can be usedas criteria for faster drying thereby maintain final productquality (Swain et al. 2012). Similar results were observed with

the combination of pulsed electric fields and partial osmoticdehydration with sucrose–salt before air drying that enhancedthe mass transfer rates and preserved the color quality of redCapsicum spp. (Ade-Omowaye et al. 2003).

In intermediate humidity products, such as pickled, theosmotic pretreatment with sucrose–salt reduced the dryingtime to obtain a pickled product with 45 % humidity.Moreover, the acceptability was no influenced by the treat-ment (da Silva et al. 2012). The presence of 2.5 % of NaClat pH 3.0, 40 °C and at chili/brine ratio 1:2 resulted in capsa-icin extraction up to 25 % in Habanero chili (C. chinense)(Hernández et al. 2009).

Drying

To maximize the color quality in red Capsicum spp. fruitsand to provide remunerative profits, it is necessary toharvest at maturity stage, when completely red fruit coloris achieved (Krajayklang et al. 2000). There are manytechniques to improve the shelf life of a product as theuse of refrigeration, freezing as well as modified

Table 3 Osmotic dehydration treatments used in Capsicum spp.

Species Common name Treatment conditions Osmotic solution Fruit: solutionratio (w/w)

Reference

C. annuum Red paprika 25 °C5 h

Sucrose (5 to 45 %) andNaCl (0 to 15 %)

1:25 Ade-Omowaye et al. 2002

C. annuum L. Red bell pepper 30 °C, 30, 60 min Sucrose: NaCl (21.86:2.02 ) 1:10 Ade-Omowaye et al. 2003

Moema 25 °C90 min

Sucrose /NaCl 1:3 (w/w) 1:4 Da Silva et al. 2012

C. annum

C. frutescens

Rodo (hot)Tatase (bell)SomboBawa

20, 30, 40 °C9 h

Sucrose (40, 50, 60 brix)Sucrose 50 brix + NaCl

(5, 10, 15 %)

1:10 Raji Abdul Ganiy et al.2010

C. chinense Habanero 40, 60 °C18 min

NaCl (7.5, 5, and 2.5 %) 1:21:31:4

Hernández et al. 2009

C. annuum L. Maras (red pepper) 25, 35, 45 °C30, 60, 90, 120 min

Common salt (20 %, 30 %,saturated)

Sea salt (20 %, 30 %, saturated)

1:20 Levent and Ferit 2014

C. chiense Moema 25 °C, 90 min Sucrose: NaCl 1:3 1:4 da Silva et al. 2012

C. annuum

C. frutensces

RodotatasheShomboBawa

9 h30 °C

Sucrose (40, 50, 60 brix) – Falade and Oyedele 2010

C. annum Jupiter (green bell) 20. 25, 30. 35, 40 °C36 to 600 min30 °C, 240 min

NaCl (0–10 g) and/orsorbitol ( 0–10 g)

5.5. g NaCl +6 g Sorbitol

1:3 Ozdemir et al. 2008

– Fresh green bell pepper 20, 35, 50 °C15, 30, 60, 90, 120,

1200 min

NaCl (2–10 %) and /orSorbitol (0–10 %)

1:3, 1:6, 1: 4.5 Ozen et al. 2002

C. annum L. Verdel (green bell) 6 to 60 minRoom temperaturePressure (4.05, 44.66

and 85.33 kPa)

Sorbitol (7, 24, 41 brix) orNaCl ( 30, 40, 110 g/L)

1:4 Quintero-Chávez et al.2012

C. annuum L. Fresh chili 30 °C, 8 h 10 % NaCl +50 % sucrose 1:10 Zhao et al. 2013

Food Bioprocess Technol (2017) 10:51–76 55

atmospheres. However, drying is still the most commonpreservation method for most vegetables and spices.Drying has been successfully applied to decrease physi-cal, biochemical, and microbiological deterioration offood products due to the reduction of moisture contentto an appropriate level, which allows safe storage over along period which results in substantial reduction inweight and volume, minimizing packing, storage, andtransportation costs (Doymaz and Kocayigit 2012).However, drying is one of the most time and energy-consuming processes in the food industry. The quality ofdried Capsicum fruits is assessed by a wide number ofdifferent parameters such as color, hotness, ascorbic acidcontent, and volatile flavor compounds (Toontom et al.2012). To accelerate the drying process with the aim ofimproving the final quality of the dehydrated final prod-uct, mostly in color terms, some pretreatments can beimplemented both in sun and industrial drying.

Between these treatments, the use of chemical pretreat-ments and osmotic dehydration seen above is the most com-mon. Usually, the term drying is used for drying under theinfluence of non-conventional energy sources like sun andwind, and dehydration is considered the process of removalmoisture by application of artificial heat under controlled con-ditions of temperature, humidity, and air flow.

Drying involves heat and mass transfer that results in re-versible and irreversible changes (either physical or as a resultof chemical or biochemical reactions) in the product (Tunde-Akintunde and Afolabi 2010). Drying temperature as well asthe drying method used influence different factors such as thedrying rate, the drying time, and the effective moisture diffu-sivity. Usually, there is an inverse relationship between airtemperature and drying time (Kooli et al. 2007). Increaseddrying temperature results in reduced drying times and ratesas well as increase moisture diffusivity, but in worse qualityfinal products (Tunde-Akintunde and Afolabi 2010). In gen-eral, the quality characteristics of dried Capsicum spp. dependon cultivar, maturity, and storage conditions of the fresh fruitsas well as the drying method used (Topuz et al. 2009). Energyconsumption and quality of dried products are critical param-eters in the selection of drying process. Moreover, to reducethe energy utilization and operational costs in the process, it isnecessary and appropriate election of the drying technique.Among the technologies, sun drying, hot drying, and freezedrying have great scope for the production of quality driedCapsicum spp.

Sun Drying

Capsicum spp. fruits on harvesting have moisture content of65–80%, depending on whether partially dried on the plant orharvested while still succulent; this must be reduced to 10% toprepared dried spice. Traditionally, this can be achieved by

sun-drying fruits immediately after harvesting without anyspecial form of treatment. In the traditional open sun drying,the product is exposed directly to the sun allowing the solarradiation to be absorbed by the material. It is one of the mostpracticed, traditional, and oldest methods employed for foodpreservation. This technique requires an area with a large,open space, and long drying times (usually more than 10 days)(Elkhadraoui et al. 2015). It is hardly dependent on the abilityof sunshine, and it is susceptible to contamination with foreignmaterial, insects, and fungal infestations, resulting in low-quality products (Fudholi et al. 2013; Topuz et al. 2009).This low quality includes red color fading, development ofbrowning pigment, and loss of carotenoids (Vega-Gálvezet al. 2008). However, the sun’s free energy for drying in openair is counterbalanced by a multitude of disadvantages whichreduce not only the quantity but also the quality of the finalproduct. To improve dried chili quality, some mechanical andsolar dryers have been introduced for drying chili in order todecrease drying time (Vega-Gálvez et al. 2008). Betweenthese technologies, there are promising applications of prom-ising solar energy systems. The mild temperature mediates theselection and proliferation of microbiota that may contributeto enzymatic PG activity modifying the pectin fraction. Theenzymatic activity generates rises in the calcium pectate frac-tion which favors the drying of the fruit with an initial lowcontent of soluble pectins and calcium pectate. The changes intexture help during the transfer of moisture facilitating thedehydration process helping bioactive compounds such as ca-rotenoids and capsaicinoids to remain almost unaltered. Onthe other hand, when the soluble pectins increases during de-hydration, the process is delayed negatively affecting the ca-rotenoid content, responsible of red color of the fruits(Gallardo-Guerrero et al. 2010).

Recently, attempts have been made to develop solarequipment’s to improve upon the sun-drying techniqueswhich lead to better use of available solar radiation, re-duction in drying time and cleaner and better qualityproduct, free from dust, dirt, and insect infestation.Greenhouse dryers showed that the moisture contentedcould be reduced to 16 % in 17 h instead of 24 h fortraditional open sun dryers with a rapid payback of theinvestment (Elkhadraoui et al. 2015). A solar drying byforced convection was used to dry C. annum resulting in a49 % saving in drying time compared with open sun dry-ing (Fudholi et al. 2013). The use of solar tunnel dryersgave acceptable moisture content and capsaicin recoveryin C. frutenscens (Yaldiz et al. 2010). Finally, a studycomparing a sun traditional oven (50 and 70 °C) andmicrowave oven (210 and 700 W) drying behaviors ofred bell pepper slices concluded that both the method aswell as the temperature had a significant effect on themoisture loss rate being microwave the best method(Arslan and Özcan 2011).

56 Food Bioprocess Technol (2017) 10:51–76

Conventional Drying

Hot air drying is probably the most popular technique fordrying Capsicum spp. in developed countries due to this rel-atively short drying time, uniform heating, and hygienic char-acteristics of the final product compared with sun drying.Usually, temperature ranges from 45 to 70 °C reducing dryingtime to less than 20 h. This temperature range provides max-imum color values and minimizes the loss of volatile oils anddiscoloration (Arslan and Özcan 2011). However, hot air dry-ing causes structural and physicochemical damages for theoverheating during the second stage of drying as a result ofshrinkage phenomenon which is taken place during the dryingprocess. A lab-scale tray dryer using a one-temperature regime(50, 60 and 70 °C) provided a darker color for dried chili andlow values of lightness (L*), chroma (C*), and hue angle (H*)compared with those when a two-stage temperature processwas used (Arslan and Özcan 2011).

Drying influences the chemical composition of green pep-pers (C. annuum L.) due to volatilization of some compo-nents, oxidation processes, and protein denaturalization, buttemperature does not have a significant role in this phenome-non (Faustino et al. 2007). The radical scavenging activityshowed higher antioxidant activity at high temperatures (80–90 °C) respect to low temperatures (5–70 °C), and the ASTAcolor was affected by temperature and presented the lowestcolor value at 90 °C. Moreover, all chromatic parameters (L*,a*, b*, C*, and H) were affected by temperature. In addition,the development of the Maillard reaction which occurs inassociation with other events could contribute to color andantioxidant capacity (Vega-Gálvez et al. 2009).

Other Drying Techniques

Freeze-drying supports enzyme deactivation, thus rapidlyinhibiting enzymatic oxidation offering a superior productquality. However, the final products result strongly hygro-scopic that may adversely affect the moisture content andthe water activity during storage. Besides, the freeze-dryingincreases the porosity and consequently the surface area of theproduct is highly exposed to the damaging activity of freeradicals and oxygen. However, this sensitivity to the processdepends on the freeze-drying conditions, the variety ofCapsicum spp., and the part of the fruit processed (Materska2014). Furthermore, freeze-drying of C. annuum Linn. Var.acuminatum Fingarh resulted in more bright red color thanother drying methods without affecting the capsaicin concen-tration and with a similar sensorial profile than fresh chili.However, in spite of its great quality, freeze-drying is still avery expensive method to implant at industrial level (Toontomet al. 2012).

Instant Controlled Pressure Drop stops the thermal degra-dation and induces swelling and possibility the rupture of the

cell walls. This technique can be used as an alternative withhigh-quality final product decreasing the operation cost(Téllez-Pérez et al. 2012) with a direct impact on active mol-ecules and functional activity in green C. annuum L. (Poblanopepper). Moreover, results showed that the process could pre-serve the main nutritional and sensorial characteristics of theraw material as phenolics and flavonoids (Téllez-Pérez et al.2013).

Reflectance Windows™ drying helps to prevent qualitydegradation of the product. In the case of Capsicum spp., thisseems to be a promising processing method showing a goodoverall color quality, except for the ASTA values comparablewith freeze-dried samples (Topuz et al. 2009).

Finally, far-infrared radiation has significant advantagesover conventional drying such as higher drying rate, energysaving, uniform temperature distribution, and reduced dryingtime, giving a better quality product (Saengrayap et al. 2015).

Conventional Extraction Procedures

Water Extraction

Water, used as an extraction solvent, mainly extracts the hy-drophilic compounds present in vegetable materials. It is asafe and cheap solvent widely used mainly in cooking andin the preparation of water infusions. Moreover, in somecases, water can enhance the nutritionally value of somespices. Water can be added hot (boiling point) or cold (at roomtemperature) to the drying plan extracts refluxing for a periodof time. After that, extracts are filtered and/or centrifuged. Insome cases, the extracts can be freeze-dried before being an-alyzed (Aliakbarlu et al. 2014; Gonçalves et al. 2013).

The traditional water extraction techniques usually includemaceration with or without stirring, mild heating, or heatingunder reflux. It is the simplest and oldest extraction method.This technique requires generally long extraction times andlarge amounts of sample and water. The most traditional useis as infusions and the hydro-distillation (El Asbahani et al.2015). However, the heating process may destroy thermal-sensitive compounds. To improve the efficiency, ultrasound-assisted extraction, pressurized hot water extraction, negativepressure cavitation-assisted extraction, and pulsed electricfields have been used to assist the process (Meng andLozano 2014). In general, the inedible portions of the fruitsare removed from the edible portion and washed in distillatedwater. Then, fresh fruits can be previously dried and powderedor not. The aqueous extract is prepared homogenizing the fruitin distillated water, and it can be boiled (100 °C, 10 min) ornot and then centrifuged to obtain the supernatant. The super-natant is usually cool, filtered, and freeze-dried prior to use. Atechnique of simultaneous steam distillation and solvent ex-traction in a Likens-Nickelson apparatus has been used to

Food Bioprocess Technol (2017) 10:51–76 57

determine the capsaicinoids content in a C. chinense Jackcultivar (Habanero) (Pino et al. 2007). Water, which is a verypolar solvent, has a poor extraction capacity of capsaicinoidsthat are non-polar. This reduced effectiveness is accentuated inthe case of the less polar capsaicinoids such asdihydrocapsaicin, homocapsaicin, and homodihydrocapsaicin(Barbero et al. 2008).

Distillation with water is, despite its low efficiency, thesimplest form to obtain an essential oil. The essential oil formsan azeotropic mixture with water. Subsequently, after the con-densation, they can be separated easily by decantation. Thesample is exposed to temperatures close to 100 °C which canlead to changes in thermolabile compounds. Prolongedheating in contact with water can lead to hydrolysis of esters,polymerization of aldehydes, or decomposition of other com-ponents, reducing the quality in the final product (El Asbahaniet al. 2015). As in almost all the extraction methods, the com-pounds extracted and the antioxidant activity obtained dependon the extraction time and the temperature (Stanojević et al.2015; Sintim et al. 2015). When water distillation has beencompared with other water extraction procedures, such assubcritical water extraction, results showed better extractingvalues with subcritical water (Gahungu et al. 2011).

Due to extremely high temperatures, steam distillation un-der reduced pressure (0.0125 MPa) at 55 °C for 4 h has beensuccessfully used to avoid production of artificial reactionflavor compounds (Jang et al. 2008) (Table 4).

Pressurized hot water extraction (PHWE) is an attractiveand environmentally friendly alternative for extraction since,at elevated temperatures, the viscosity and high surface ten-sion of water decrease, while diffusivity and solubility of thecapsaicinoids increase. However, the PHWE requires a so-phisticated instrumentation since it needs an application ofhigh pressures and temperatures (Bajer et al. 2015).However, this method has resulted more efficient to extractcapsaicinoids than traditional Soxhlet extraction using metha-nol as solvent (Bajer et al. 2015).

Maceration

Mixing of a solvent and a solid matrix is one of the oldest andsimplest technologies to transfer some compounds from afood matrix (in this case the fruit) to a solvent bulk. The plantextracts can be prepared directly from fresh fruit or from pre-viously dried fruits. To avoid thermal degradation, macerationis carried out at room temperatures or under vacuum. Thebasic procedure consists of transferring the sample into thesolvent for a long period, protected from light and with onedaily agitation. Themost common solvents used are methanol,ethanol, water or a mixture of them (Sasidharan et al. 2011; deAguiar et al. 2015) although maceration with vegetable oils isincreasing. More expensive methods such as homogenizationwith liquid nitrogen (Jang et al. 2008) and carbonic

maceration (Liu et al. 2014) have been used at experimentallevel. (Table 4). Generally, after maceration, the solid materialdissolvent in the solvent phase is separated by filtration orcentrifugation.

Olive Oil Maceration

The addition of aromatizers to the olive oil influences severalcharacteristics and properties. Their inclusion improves oliveoils sensorial characteristics, but concentration must be kept atlow or moderate levels in terms of sensorial acceptability byconsumers to avoid over-aromatization (Sousa et al. 2015).There is an increasing demand for top quality, healthy, andinnovative food products. Consequently, flavored oils andoil macerated extract methodologies have been increasing de-veloping. Flavored extra-virgin olive oil is the most aromaticoil commercialized. It could be done with essential oils, fruits,herbs, mushrooms, spices, and vegetables. These flavoringscould be added to the olive oil before or after the oil extraction(Sousa et al. 2015).

Cold pressing process, used to obtain extra-virgin olive oil,has been analyzed in two ways to flavor the final product:mixing the Capsicum seeds or fruits with olives before thepressing to obtain the flavored olive oil or by cold pressingof the Capsicum seeds. In the first case, the extraction is madeof the mixture of olives and Capsicum powder and the yieldand functional properties (as antioxidant) of this extraction islow (Baiano et al. 2009). In the second case, seeds are coldpressed alone to obtain the oil but the yield and the consumeracceptance are low for the high pungency of the oil (Yılmazet al. 2015). The production of flavored oils by maceration fora long time period (between 1 and 3 months) in darkness andat room temperature is the most common technique to con-serve the nutritional and bioactive quality of oils (Caporasoet al. 2013; Sousa et al. 2015; Ciafardini et al. 2006; Ciafardiniet al. 2004) (see Table 4).

In order to accelerate the olive oil aromatization, new tech-niques have been developed as the use of supercritical fluidextraction (SFE) with supercritical CO2 to extract oil from redfruits of Capsicum spp. The Capsicum oleoresin was thenadded to an extra-virgin olive oil and its oxidative stabilitywas evaluated. Results showed that the Capsicum oleoresinproduced by SFE can be used to produce stable flavored oliveoils (Gouveia et al. 2006). Other accelerated methods such asultrasound-assisted extraction (UAE) (10–20 % w/v, 20 min)and microwave-assisted extraction (MAE) (10–20 % w/v, 1–60s) have been employed using 10–20 % of red chili powder.The content of capsaicinoids extracted by traditional infusionwas higher than the non-conventional techniques. However,the production of flavored olive oils using technologies suchas ultrasounds and microwaves could allow the production ofhigh quality oils with an important time reduction (Paduanoet al. 2014).

58 Food Bioprocess Technol (2017) 10:51–76

Vegetable Oil Maceration

Themaceration of dehydratedCapsicum spp. in vegetable oilshas been recently carried out using some comestibles (olive,corn, sunflower, safflower, coconut, and palm) and medicinaloils (mustard, neem, ricinums, ground nut, and ginger) at dif-ferent temperatures and extraction times and then filtered andcentrifuged to separate the solids to obtain the non-aqueousextracts of chili (Guadarrama-Lezama et al. 2012; Amruthraj2014). The extraction of components from C. chinense (BhutJolokia) in different vegetable oils depends on the solubility ofcapsaicin and other compounds present in the oil, and it wasalso an efficient solvent to extract capsaicinoids for biologicalapplications (Amruthraj 2014).

Vegetable oil extracts of C. annuum L. showed high anti-oxidant activity without the use of organic solvents potentiallyharmful to the environment. Results depended on the extrac-tion conditions and the refined vegetable oil employed so that,in general, lower temperatures resulted in higher antioxidantactivity and the best vegetable oil was corn oil due to itsparticular fatty acids profile (Guadarrama-Lezama et al.2012) (see Table 4).

The use of ozonated oil has been developed with industrialpurposes. However, this oil must not be consumed because thehazards related with ozone. Basically, the mixture with thevegetable oil and the chili powder is bubbled with oxygengas and ozone until a grease with antimicrobial properties isobtained (Özyildiz et al. 2013).

Cold Pressing

Cold pressing is the most antique process to obtain oilsconsisting only in a mechanical pressing without heating.

This method has the advantage or little or no heat generationduring the process giving good quality products that do notrequire refining but gives low yields (Reyes-Jurado et al.2015). It has traditionally been employed to obtain virgin oliveoil and in the last years to produce avocado oil and other oilsseeds (Dos Santos et al. 2014; Febrianto and Yang 2011). Inthe case of essential oils, it has been traditionally used toextract essential oils from citrus fruits. During extraction, oilsacs break down and release volatile oils. Then the oil is re-moved mechanically by cold pressing yielding a watery emul-sion. Oil is recovered by centrifugation (El Asbahani et al.2015). Capia pepper seed oil was produced by a cold pressingtechnique; however, the extraction yield was very low com-pared with the traditional Soxhlet extraction with hexane andthe resulted oil was not accepted by consumers (Yılmaz et al.2015).

Organic Solvent Extraction

The plant material is macerated in an organic solvent; theextract is usually concentrated by removing the solvent underatmospheric or reduced pressure (El Asbahani et al. 2015).Manymaterials including oils, fats, and proteins are recoveredfrom diverse biological sources by this type of extraction. Thebioactive compounds in Capsicum fruits vary in chemicalstructure between cultivars; the solvent characteristics have astrong effect on the compounds present in the extract and theirtested activity (Bae et al. 2012b) (see Table 5).

Experimental data indicates that the concentration and ac-tivity of bioactive compounds in natural foods may be directlyrelated to solvent properties such as lipophilic and hydrophilicsolvents and their respectively polarity. Hence, lipophilic ca-rotenoids are better extracted in non-polar solvents, such as

Table 4 Various experimental conditions to obtain Capsicum spp. oleoresins by non-conventional maceration

Maceration type Common solvent used Temperature (°C) Pressure (MPa) Time required Reference

Solvent maceration Liquid nitrogen to preventthe loss of volatiles(pretreatment)

Deionized water (steamdistillation underreduced pressure)

55(steam distillation

under reducedpressure)

0.0125(steam distillation

under reducedpressure)

4 h(steam distillation

under reducedpressure)

Jang et al. 2008

Solvent maceration Carbonic maceration 30 0.2000 30 h Liu et al. 2014

Flavored olive oil Olive oil Room temperature Room pressure 3 months Sousa et al. 2015

Oil infusion Olive oil Room temperature Room pressure 7–30 days Caporaso et al. 2013;Paduano et al. 2014

Flavored olive oil Olive oil Room temperature Room pressure 14 days Ciafardini et al. 2004

Non-aqueous extracts Corn, Sunflower andsafflower oils

60–70-80 Room pressure 5–10 min Guadarrama-Lezamaet al. 2012

Extraction invegetable oils

Mustard, sunflower, olive,coconut, palm, castor,neem, ground nut andgingelly oils

65 Room pressure 1 h Amruthraj 2014

Food Bioprocess Technol (2017) 10:51–76 59

hexane, but flavonoids being hydrophilic are easily extractedin polar solvents as water (see Table 5). However, the antiox-idant activity of each extract does not represent the totalamount of bioactive compounds present in the whole fruitsince the fruit has a wide variety of compounds with variouspolarities and they cannot be extracted totally by only onesolvent (Bae et al. 2012b). There are many physical and chem-ical differences between the diverse biological materials.However, oils (edible and industrial) and other useful lipidicmaterials (vitamins, nutraceuticals, fatty acids, phytoesterols,etc.) can be extracted by mechanical pressing, solventextracting, or a combination. The preparation of the materialto be extracted differs, but generally cleaning and drying aresteps required. For solvent extraction, the plant material isdissolved in a solvent to separate the oil from the insolublemeal. Many solvents have been evaluated for commercialextraction.

In a typical chemical extraction process, solvents are usedfor dissolving reactants, solvating molecules, extracting prod-ucts, and separating mixtures. However, the major part of theorganic solvents currently found in industry is characterizedby several dangerous effects for the human health and envi-ronment (Vian et al. 2014). The solvent extraction is the mostcommonly used technique to obtain oleoresins fromCapsicum spp. usually used as color additive. The currentEuropean regulation (Commission regulation N° 231/2012)allows the use of ethyl acetate, methanol, ethanol, acetone,hexane, and isopropanol with a solvent residue of not morethan 50 mg/kg individually or in combination and no more of10 mg/Kg of solvent residue in dichloromethane. Meanwhile,FDA (2006) allows the use of acetone, ethanol, ethylenedichloride, hexane, isopropanol, methanol, methylenechoride, and trichloroethylene. The extracted profile obtaineddepends on the solvent polarity and other physico-chemicalproperties of the particular solvent (see Table 5) and from theextraction conditions (time and temperature) (see Table 6).Therefore, less polar solvents as hexane, acetone, and metha-nol can extract carotenoids easily (Arimboor et al. 2015) andare less recommended to extract capasaicinoids than polaraprotic solvents such as acetone and acetonitrile and evenfor polar protic solvents such as methanol and ethanol(Amruthraj 2014).

At high temperatures and long times, some reactions canoccur and extraction rates of some compounds can decrease(El Asbahani et al. 2015). Solvent polarity could significantincrease the extraction efficiency of specific lipophilic or hy-drophilic compounds in different peppers as well as antioxi-dant activity. Hexane and methanol have a fairly narrow boil-ing point that makes them easy to recover (Yılmaz et al. 2015;Wang and Weller 2006). However, both are considered toxicsfor human health and for the environment and their presencein food is strictly regulated (with a maximum allowed of290 ppm) (de Aguiar et al. 2013; Fernández-Ronco et al. 2013;T

able5

Solventcharacteristicsandrelativ

epolarity

ofcommonly

used

solventsin

Capsicumspp.extractio

n.Adapted

from

Jouybanetal.2011andReichardt

2004.

Solvent

Meltin

gpointat

0.1013

MPa

(°C)

Boilin

gpointat

0.1013

MPa

(°C)

Flashpointat

0.1013

MPa

(°C)

Refractiveindex

at25

°CAbsoluteviscosity

at25

°C(cP)

Density

at25

°C(g/cm

3 )So

lubilityin

water

at25

°C(%

w/w)

Relativepolarity

n-hexane

−95

69−22

1.372

0.31

0.659

9.5×10

−4

0.009

Diethyl

ether

−116

34.5

−45

1.352

0.24

0.715

6.9

0.117

Ethyl

acetate

−84

83−4

1.375

0.72

0.867

7.7

0.228

Methylene-chloride

−95

40Would

notflash

1.421

0.44

1.326

1.30

0.309

Acetone

−95

56−18

1.357

0.33

0.79

Total

0.355

Acetonitrile

−44

81.6

61.342

0.38

0.782

Total

0.460

Isopropanol

−89

82.6

11.7

1.377

2.1

0.785

Total

0.546

Ethanol

−114

7813

1.359

1.08

0.789

Total

0.654

Methanol

−98

6415

1.326

0.6

0.792

Total

0.762

Water

0100

Would

notflash

1.332

0.89

0.998

Total

1.000

60 Food Bioprocess Technol (2017) 10:51–76

Menchini et al. 2009). Other organic solvents used to obtainCapsicum oleoresins are acetone, acetonitrile, isopropanol,and ethanol (Table 6) (Dorantes et al. 2000; Chinn et al.2011; de Aguiar et al. 2014). However, besides the organicsolvent used, the capsaicinoids yield depends on the part ofthe fruit examined. There are few solvents that can be useddirectly to obtain carotenoids and capsaicinoids from freshfruits (high moisture samples). These solvents should be hy-drophilic such as methanol, ethanol, and acetone (Dong et al.2014). Nowadays, the use of non-polluting and non-toxicsolvents is imperative in most countries, so solventbyproducts from petroleum are being substituted from lesstoxic alternatives, such as ethanol, particularly in productsfor human consumption (Arimboor et al. 2015). Hence, aJapanese research of the organic solvent residues levels from145 food additives, 23 health food materials, and 19 com-mercial health food products found that ethanol was thedominant commercial solvent followed by methanol, acetone,isopropanol, and ethyl acetate. In this study, three samples ofcommercial paprika oleoresin were analyzed showing thattwo of them were extracted with ethanol and one with meth-anol (Uematsu et al. 2008).

N-Hexane is the most traditional solvent for oil extrac-tion (Table 6). It is a non-polar solvent mostly used forthe extraction of vegetable oils. Hexane has a fairly nar-row boiling point range of approximately 63–65 °C, and itis an excellent oil solvent in terms of oil solubility andease of recovery and recycling. However, acute inhalationexposure of humans to high levels of hexane causes mildcentral nervous system effects and chronic exposure isassociated with polyneuropathology (Wang and Weller2006). Hexane leads to the extraction of dyes and sub-stances (carotenoids) in Capsicum oleoresins and showshigher extraction yield than ethanol in some studies ofconventional solvent extraction due to the low relativelypolarity of the solvent (Fernández-Ronco et al. 2013).Typically, dried red Capsicum is extracted as a mash ina large heated volume of n-hexane; the extracted liquid isrecovered and the hexane is evaporated or distilled fromthe sample leaving an oleoresin and recovering the sol-vent. However, capsaicinoids are partially soluble in hex-ane. For this to obtain non-pungent Capsicum oleoresins,only mild or non-pungent fruits can be used (Richins et al.2010). Finally, conventional extraction with n-hexane isused as a solvent in non-conventional methods such asMAE (Williams et al. 2004), UAE (Fernández-Roncoet al. 2013) and SFE (Duarte et al. 2004), although withless favorable results to extract capsacinoids in compari-son with other more polar solvents such as acetone.

Acetone and methanol are used both to extract carot-enoids (Hornero-Méndez and Mínguez-Mosquera 2001)and capsaicinoids (Amaya-Guerra et al. 1997; Amruthraj2014) for their medium relative polarity. However, atT

able6

Someexperimentalconditio

nsto

obtain

Capsicumspp.oleoresins

byorganicsolventextraction

Solventu

sed

Extractiontype

Temperature

(°C)

Tim

erequired

Sample(g)/solvent

(mL)ratio

Reference

Ethanol

(70%)

Maceration

7030

min

1:10

Gao

etal. 2005

Ethanol

n-H

exane

Methanol

Maceration

Not

specified

Not

specified

Not

specified

Menichini

etal. 2009

Ethanol

Twostages

extractio

n40–50

%ethanolatroom

temperature

95%

ethanolat9

0°C

30min

follo

wed

by120min

1:10

followed

by1:4

Dongetal.2014

Ethanol,acetone,acetonitrile

Macerationwith

shaking

501h

1:6.6

Chinn

etal.2011

Hexane

Soxhletextraction

696h

1:30

DeAguiaretal.2013

Hexane,ethanol

Maceration

25–65

48–72

h1:1

Fernández-Ronco

etal.2012

Hexane,ethanol,acetone

andmethanol

Maceration

Untilboiling

6h

1:30

DeAguiaretal.2014

Hexane,ethylacetate,acetone,

methanol,methanol/w

ater

(80:20,v/v)

Soxhletextraction

6010

h1:25

Bae

etal. 2012a,b

Isopropanol

Macerationwith

shaking

Room

temperature

15min

1:1

Dorantesetal.2000

Methanol

Maceration

Room

temperature

144h×3tim

es1:10

Confortietal.2007

Methanol,ethanol,or

mixture

ofalcoholand

water

Maceration

Depending

thesolventu

sed

1h

1:1

Sasidharan

etal.2011

Food Bioprocess Technol (2017) 10:51–76 61

industrial level, these solvents are not commonly usedbecause their toxicity and flammability. Acetone is con-sidered the best polar aprotic solvent for the efficient ex-traction of capsaicinoids for pharmacological and biolog-ical purposes (Amruthraj 2014). Acetonitrile is notallowed in foods, but it has been used to extractcapsaicinoids and carotenoids for scientific purposes (deAguiar et al. 2013; Al Othman et al. 2011; Arimboor et al.2015). Isopropanol was used to obtain lipidic extracts ofCapsicum with antimicrobial properties (Dorantes et al.2000). It has also been used combined with n-hexane todetermine tocopherols in Capsicum spp. a (Abbeddouet al. 2013) and capsaicinoids by HPLC (Caporaso et al.2013) and with methanol–water to determine carotenoids(Daood et al. 2002).

Finally, ethanol is the most used organic solvent in the foodindustry because of their low toxicity both to humans andenvironment (Singh et al. 2014). Although the extraction yieldof carotenoids is not higher, some authors have reported agood extraction yield of capsaicinoids with ethanol/watermixtures both in maceration (Amaya-Guerra et al. 1997; AlOthman et al. 2011) and Soxhlet extraction (de Aguiar et al.2014). However, to extract color pigments (carotenoids), othersolvents such as acetone (Chinn et al. 2011) or hexane(Fernandez Ronco et al. 2012) provided oleoresins with bettercolor quality.

Because of their low toxicity, ethanol has been widelyresearched as an organic solvent in Capsicum extractions.Therefore, despite not having the best extraction yield, ithas been optimized in many researches. Therefore, Gaoet al. 2005 tried to optimize the process of extractingcapsaicinoids in C. annuum L. with a mixture of 70 %ethanol/water at 70 °C, 0.5 h and a stock ratio 1:10(mass/volume) in a three-step extraction. Differentethanol/water solutions have been used to extract selec-tively carotenoids and capsaicinoids in dried hotCapsicum spp. (Santamaría et al. 2000) and fresh hotred Capsicum (Dong et al. 2014) with great yield rates.Most of the solvents are employed to extract from previ-ously dried fruits in order to avoid the water oxidationand to improve the solubilization of the lipophilic sub-stances. However, there is interest in the development ofextraction methods with fresh fruits in order to reduce thetotal costs of the process and to improve the colorantcapacity since carotenoids may be partially destroyed dur-ing the drying process. The first step with a 40–50 %ethanol in fresh fruits (with high water content) was usedto extract most of capsacinoids and the second step with95 % of ethanol was used to extract carotenoids by Donget al. 2014. In the case of dried Capsicum (Santamaríaet al. 2000), the extraction of capsaicinoids was per-formed with a 30 % ethanol solution followed by a carot-enoids extraction with a 96 % ethanol solution.

Emergent Technologies for Extraction

There is an increasing demand for new extraction techniqueswith the properties of amenable to automation, shortened ex-traction time, and reduced organic solvent consumption whichnot only prevents pollution but also reduces sample prepara-tion costs. (Yu et al. 2009). Between them, microwave-assisted extraction (MAE), ultrasound-assisted extraction(UEA), and supercritical fluid extraction (SFE) are the mostimportant technologies developed during the last years.

Microwave-Assisted Extraction

MAE is mostly used to extract water from the particles todry them or as a pretreatment before or at the same timethat conventional dehydration processes. For example, it iscommonly used in combination with air-drying systemduring drying increasing the drying rate of the final prod-uct and the quality of the dried product obtained (Arslanand Özcan 2011; Swain et al. 2012). However, MAE effi-ciency and selectivity depend significantly on the dielectricconstant of the extraction solvent mixture, namely itschemical polarity that defines the compounds extracted(Csiktusnádi Kiss et al. 2000).

The MAE process allows higher recovery of analytes andreproducibility than conventional techniques. Also, it is aneasy to use and relatively low-cost technology (Yu et al.2009). Many MAE techniques have been developmental fornatural products such as open system microwave heating withwater or organic solvents, compressed air microwave distilla-tion, vacuummicrowave hydrodistillation, solvent-freemicro-wave extraction, and microwave hydrodiffusion and gravity(Mason et al. 2011). However, in Capsicum spp. extraction,the most used technique is the MAE extracted with hexane asorganic solvent where the fruit is immersed in a non-absorbingmicrowave solvent such as hexane and irradiated by micro-wave energy (Gogus et al. 2015). Here, the MAE is an alter-native extraction method because of their considerable savingin processing time, solvent consumption, and energy having ahigh extraction rate of volatile compound in Capsicum spp.compared with other traditional techniques and even withMAE extracted with water as solvent, probably because thelow solubility of volatile compounds (Gogus et al. 2015).However, the severe thermal stress and localized high pres-sures produced during MAE cause the cell rupture more rap-idly than in conventional extraction. Because of that, it isconvenient that volatile oils are dissolved in the organic sol-vent before being separated by liquid–liquid extraction(Mason et al. 2011; Gogus et al. 2015). Normally, the sample(between 1 and 10 g) is immersed in non-microwave absorb-ing solvent (such as hexane) for 3–30 min, and then micro-waves are applied into a low volume of solvent (aprox.40 mL). To avoid these drawbacks, recently, microwave-

62 Food Bioprocess Technol (2017) 10:51–76

vacuum drying and far infrared-assisted microwave vacuumthat are a free solvent extractions have been successfully usedfor drying red Capsicum spp., but not to extract its compo-nents. In MAE extraction, no degradation of capsaicinoidsfrom fresh samples of Capsicum spp., at laboratory level,was observed up to 150 °C being a faster method comparedwith traditional ones and the better parameters to extractcapsaicinoids have been the following: 125 °C, 500 W,25 mL of 100 % of ethanol as solvent, 0.5 g of triturated freshfruits, and 5 min of extraction time (Barbero et al. 2006).Similar results were obtained using acetone at 30 % wherethe extraction time was considerably reduced (Williamset al. 2004). However, in a research to extract capsaicin fromC. frutescens with different methods, MAE showed a lowerextraction recovery probably because the lack of proper stir-ring of the solvent during the extraction process (Nazari et al.2007). Microwave vacuum drying was initially developed toincrease the extraction yield of oxidable components such asantioxidants, which are important bioactive compounds inCapsicum spp. in this technology, the extraction can be per-formed at lower temperature and the air in the extraction sys-tem is mostly pumped out, so the oxidation is avoided orreduced since there is little oxygen in the process of extraction(Yu et al. 2009). Far infrared could be used as an additionalenergy source to assist microwave-vacuum drying improvingthe drying rate and giving a better quality product with lowerchanges of all color parameters, shrinkage coefficient, andhardness while rehydration ability was found higher.Consequently, the optimum condition for drying Capsicum

spp. was a microwave power of 300 W under absolute pres-sure of 21.33 kPa with the applied far-Infrared power of300 W (Saengrayap et al. 2015).

Finally, osmotic dehydration has been successfully used asa pretreatment before MAE for faster drying, thereby main-taining final product quality (Swain et al. 2012).

Ultrasound-Assisted Extraction

Ultrasounds can enhance the extraction process by increasingthe mass transfer between the solvent and plant material. Inaddition, the collapse or cavitation bubbles leads to better celldisruption via the formation of microjets due to asymmetricalbubble collapse near a solid surface. This allows for improvedsolvent penetration into the plant body and can also breakdown cell walls (Mason et al. 2011). The method is inexpen-sive, simple, and an efficient alternative to conventional ex-traction techniques that increase the extraction yield, reducethe process time and the operating temperature, and even in-crease the quality of the extract (Wang and Weller 2006).However, these benefits depend on the nature of the plantmatrix. In hot varieties of C.annuum and C. chinense, UAEhas been successfully used to extract capsaicinoids usingmethanol as solvent at relatively low temperatures (50 °C)

and short time (10–15 min) and 360 W of power (Barberoet al. 2014; Barbero et al. 2008; Sganzerla et al. 2014).Ethanol (a much less toxic solvent) has also been used toextract capsaicinoids from dried C. frutescens on a pilot scale,but the yield extraction was lower than the obtained fromtraditional industrial scale maceration with organic solvents.However, the results showed that the potential use of UAEcould shorten the extraction time and lower the operating tem-perature which resulted in lower operation costs (Boonkirdet al. 2008).

This method has been successfully used to extract colorantsfromCapsicum spp. varieties obtained a high concentration incaroteonids using hexane and ethanol as solvents in contrast tothe classical process of maceration using the same solventsmostly because the extraction time was reduced (Fernández-Ronco et al. 2013).

Supercritical Fluid Extraction

SFE has been applied as an alternative to traditional methodsfor the extraction and fractionation of active compounds,mainly lipid compounds from natural matrices. Carbon diox-ide is the most common supercritical fluid employed. It be-haves a critical temperature of 31.35 °C and critical pressureof 7.39MPa expanding to fill its container like a gas but with adensity like that of a liquid. It is an appropriate solvent for theextraction of bioactive compounds from biological substratesdue to its low cost, non-toxicity, non-flammability, inertness,and good extraction capacity although limited to dissolvecompounds with high molecular weight, regardless of theirpolarity such as carotenoids. Indeed, the critical properties ofCO2 are moderated when compared with other green solventsallowing SFE to be carried out with low-cost energy for pres-surization and temperatures that do not damage the targetcompounds (Santos et al. 2015; Araus et al. 2012). The ob-tained extracts in red C. annuum L. vary from orange, lightred, to intense dark red, depending on process conditions, andhave several advantages over extracts conventionally obtainedwith organic solvents because they do not have residues oftoxic solvents and it is possible to vary the content ofcapsacinoids and carotenoids by changing the extraction pa-rameters (Perva-Uzunalić et al. 2004). Extraction using CO2 isgenerally carried out with two-stage separation of extracts intoa pungent oleoresin and an essential oil fraction. The extract isviscous, pasty, and semisolid and thus difficult to recover fromseparation vessels (Catchpole et al. 2003). The main disadvan-tage of this technology in comparison to the conventionalmethods is the high investment requirements related to thehigh pressure operation, although results have demonstratedthe economic feasibility of the process to obtain bioactivecompounds from Capsicum oleoresin (Fernández-Roncoet al. 2013). The development of commercial SFE systemsrequires the economical evaluation of the process at different

Food Bioprocess Technol (2017) 10:51–76 63

levels. At the initial stage, the cost of equipment and cost offabrication can be approximate but needs to make into accountboth the capacity and the inflation in a detailed way. In thecase ofC. chinense (Habanero) oleoresins is possible to obtainoleoresins free of organic solvents that may be used for humanconsumption without risk. Moreover, the difference betweenthe manufacturing cost and the probably selling price presentsa good perspective for industrial application (Rocha-Uribeet al. 2014). The particle size, moisture content, and oil con-tent affect the SFE. A decrease in particle size increases theextraction efficiency and moisture levels higher than 18 %decrease the efficiency (Nagy and Simándi 2008).

Fractionation of spice extracts with SFE using CO2 as sol-vent using 2, 3, or 4 separator vessels at different pressures andtemperatures to obtain fractions with different characteristicshas been reported (Jarén-Galán et al. 1999). These authorsfound that, in the case of hot fruits, an orange-yellow pungenttasting extract containing capsaicin and few carotenoids wasextracted in the first stages at lower pressures. However, in thesecond stages, at higher pressures, a darkness extract contain-ing carotenoids and fatty acid was obtained. In sweet fruits,approximately 62 % of total carotenoids in raw material wasrecovered in the extract, and the rest remained occluded in theheat exchanger (Ambrogi et al. 2002). For the analysis of shortand long chain free fatty acids from seeds of Capsicum spp.,SFE with supercritical CO2 using ethanol as co-solvent wassuccessfully employed (Li et al. 2011).

Only few researches have been reported using others ex-traction fluids, such as sub-critical propane and dimethylether. Propane is cost-effective alternative to SFE using CO2

due to the lower operating pressures and lower energy con-sumption required. Dimethyl ether has similar advantages topropane (Catchpole et al. 2003).

Results showed that subcritical dimethyl ether was as ef-fective at extracting the pungent principles as supercriticalcarbon dioxide, although a substantial amount of water wasalso extracted. However, subcritical propane was less effec-tive, obtaining a half yield of capsaicinoids than supercriticalCO2 and subcritical dimethyl ether (Daood et al. 2002;Catchpole et al. 2003). The yield of paprika extracts(C. annuum L.) was found fairly constant with subcriticalpropane at different conditions and resulted superior thanCO2 to extract carotenoids and thocopherols (Gnayfeed et al.2001).

However, the partial extraction of pungent compoundswith subcritical propane may be useful for the production ofoleoresin with slight pungency and high color intensity.Subcritical propane may be useful to extract fatty acids, to-copherols and carotenoids (Gnayfeed et al. 2001).

SFE with CO2 has been used to extract both carotenoidsand capsaicinoids from varieties of C. annum, C. frutescens,andC. chinense under different process conditions. In general,Capsicum oleoresins are mainly composed by lipid matter,

carotenoids, and capsaicinoids. Between carotenoids,capsorubin, capsainthin, zeaxanthin, β-cryptoxanthyn aremainly responsible from the red color while β-carotene actsmainly as an antioxidant. Most commercial valued oleoresinsare those that present at high red coloring capacity that iscaused by carotenoids pigment with or without pungent flavordue to the presence of absence of capsaicinoids (Fernández-Ronco et al. 2011). In few cases, tocopherols have been ana-lyzed (Abbeddou et al. 2013).

Capsicum oleoresin has one of the highest carotenoids pig-ment concentration of products derived from natural sources.The use of supercritical CO2 reduces the isomerization anddecomposition of carotenoids and increases the extractionyield because of the low polarity of these compounds(Barros et al. 2015). In non-pungent varieties of C. annuumL., the highest extraction yield was found at 60 °C, 24 MPa,and 12 min and with 0.2–5 mm of particle size. Applyingthese conditions, the extraction yield was 97.0 and 68.1 %for tocopherols and carotenoids, respectively (Romo-Hualdeet al. 2012). In pre-pelletized Jalapeño peppers (C. annuumL.), the flakes were conditioned to low moisture, ground fine-ly, and pelletized at high pressure before the SFE using CO2 assolvent. The oleoresin obtained after 4 h at 40 °C and 12 MParesulted in a light yellow tinge (del Valle et al. 2003).

A fractioned extraction has been used in other non-pungentC. annuum L. varieties using 1 % of ethanol or acetone as co-solvent to obtain highest red pigment yields. During thefractioned extraction, in the first step, at low pressure, thecompound extracted is almost exclusively β-carotene whilein the second stage, at high pressure, carotenoids responsiblefrom red color (capsorubin, capsanthin, zeaxanthin, and β-cryptoxanthyn) were mainly obtained (Jarén-Galán et al.1999). Ethanol as co-modifier and tapping solvent has alsoused in hot Capsicum varieties allowing the inclusion of adifferential precipitation step without extract of chlorophyllpigments that may present as contaminants in dried fruit sam-ples (Richins et al. 2010). Recently, avocado oil has been usedas co-solvent in simultaneous extraction using SFE to extractcapsainthin from C. annuum L. with a clean technology. Forthat, freeze-dried avocado and Capsicum fruits were separate-ly packaged in a single-bed extractor in which supercriticalCO2 was passed through the bed at 50 °C and 40 MPa. Thetarget compounds were simultaneously extracted from bothsources extracted giving intensely red-colored oil containing280–460 μg/g capsanthin with an extraction yield of around30–50 % of capsanthin was extracted (Barros et al. 2015).

The extraction of capsainthin has been successfully im-proved with the use of triolein-entrained SFE with CO2 assolvent because the triolein increases the solubility ofcapsanthin independent on system temperature and pressurebecause of their nonpolar interactions between the 18-C fattyacid chains in triolein and the 22-C nonpolar core of caroten-oids (Araus et al. 2012).

64 Food Bioprocess Technol (2017) 10:51–76

Capsaicinoids has been extracted both in hotC. annuum L.,C. chinense, C. baccatum, and C. frutescens varieties. In thecase of red hot C. annuum L. with high pressure apparatususing near and supercritical CO2 results showed that thehighest yield of oil and capsaicin content was found at 60 °Cand 35 MPa (Kwon et al. 2011). Moreover, at lab level, SFEwith supercritical CO2 has been used to extract capsacinoidswith a wide concentration range (Peusch et al. 1997; Sato et al.1999). In C. frutescens SFE with supercritical CO2 as solvent,high extraction yields in oleoresins and capsaicinoids wereobtained at pressures around 20–22MPa. The extract obtainedgave similar pungency virgin olive oil than commercial fla-vored olive oils without modification of the original oil color(Duarte et al. 2004).

In native Brazilian Capsicum spp., the highest concentra-tion of capsaicinoids was found in C. frutescens. Using super-critical carbon dioxide, the highest concentration was found atthe lower pressures, indicating that it may be a competitionbetween capsaicinoids and other soluble compounds for su-percritical CO2 at higher pressures being the best extractionconditions 15 MPa and 40 °C. Moreover, the extraction kinet-ics of capsaicinoids showed that these substances are extractedin the first minutes of extraction, when mass transfer rate ishigh (de Aguiar et al. 2013).

Recently, SFE has been successfully assisted by ultra-sounds to extract capsaicinoids from C. frutescens L. usingCO2 as solvent increasing the global yield of the oleoresin upto 30 % when compared to SFE without changing the totalcapsaicinoids and phenolics profile. The best operating con-ditions resulted 360 W for 60 min and 40 °C, 15 MPa, and1.673 × 10−4 kg/s (Santos et al. 2015). Moreover, the highestextraction rates were obtained for high CO2 flow rates, lowparticle diameter, and low extraction bed volume. These re-sults could be explained by the high importance of the con-vective phenomenon under these conditions (Silva andMartínez 2014).

The extraction of capsinoids with SFE using supercriticalCO2 has been studied in sweet C. chinense where highcapsaicinoids and low capsaicinoids concentrations are natu-rally presents. SFE process with CO2 resulted in low extrac-tion yields when compared to solvent extraction with metha-nol and acetone. However, the extract obtained with the low-est CO2 density had a high concentration of capsinoids, sug-gesting that the SFE using CO2 without co-solvent processcan be used to obtain concentrated extract of these extractsfor further application in food and pharmaceutical industry (deAguiar et al. 2014).

Pelletization can increase the volumetric yield of SFE withsupercritical CO2 at 40 °C and 32MPa, in greenC. annuum L.(jalapeño). Sample compactation by pelletization appeared tobe slightly less effective when using flakes than ground ma-terial, independent on the initial sample moisture. A reductionin the particle size of the pellets improved the mass transfer

but caused a reduction in packing density, and these two fac-tors had opposite effects on the volumetric yield of the process(mass of recovered solute per unit time and per unit volume ofextraction vessel), which was also affected by prepelletizingsample conditioning. (Uquiche et al. 2005).

Others

Application of high intensity pulsed electric field (PEF), a lowthermal short time process induces membrane perme-abilization in various biological cell systems due to electricalbreakdown of the membrane. The resultant membrane perme-abilization affects heat and mass transfer processes in subse-quent unit operations such as extraction with improved qualityof the extracted juices. InC. annum L. (paprika), an extractionyield about 91 % and good quality of juice was obtained fromthe fruit mass after a pretreatment with electric pulsed electricfields followed by pressing at 10 MPa (Ade-Omowaye et al.2001). As a drying pretreatment, PEF was applied toC. annuum L. with electric field strength of 1.0–2.5 kV/cm using a fixed pulse width of 30 μs and at a pulse fre-quency of 100 Hz. The pretreatment resulted in cell mem-brane disruption without visible changes in the surfacestructure of the fruit. The reduction in drying time byPEF pretreatment was beneficial to color quality of driedred pepper (Won et al. 2014).

Pressurized liquid extraction (PLE) is a relatively nov-el extraction method in which temperature and pressureare utilized to accelerate the extraction of compoundsoriginated from solid or semisolid samples. The applica-tion of high temperature and high pressures modifies thephysical properties of the extraction solvents, with theeffect of increasing selectivity in the extraction workingabove the boiling point of the solvent and enhances theextraction efficiency because it decreases the viscosity ofthe solvent, allowing a better penetration of solvent mol-ecules into the sample matrix. PLE uses less organicsolvent, and the extraction time is reduced comparedwith traditional techniques. However, the equipment ishighly expensive due to its precision. In addition, PLEcan be performed in the absence of oxygen and lightwhich reduce the oxidative degradation of sensible com-pounds such as antioxidants (Liu et al. 2014; Barberoet al. 2006). PEL has been used with water, ethanol, ormethanol (0–20 % in water) to extract capsaicinoids inC. annuum varieties of different pungency showing thatmethanol was the best extraction solvent (Barbero et al.2006). Finally, PEL has method has been optimized withmethanol as solvent at 100 °C and 10.3 MPa to extractcapsaicinoids from a dried C. annuum cultivar (Liu et al.2014). However, methanol is not a good solvent for thefood and pharmaceutical industry for its high toxicityand flammability (Jin et al. 2004).

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Bioactivity

Health Benefits from Capsicum spp.

Capsicum spp. are one of the most consumed vegetablesworldwide, mainly due to the diversity of culinary purposesand its handling plasticity. The consumption of both fresh anddry as spice seems to have positive effects on human health(Tundis et al. 2011). From a nutritional point of view,Capsicum spp. fruits are generally considered good sourcesof most essential nutrients. They are rich in antioxidants,colors, flavor, and vitamins such as vitamins E, A, C, and Bcomplex. The quality of the fruit depends on its chemicalcomposition. Among most important factors that affect com-position, environmental and growth conditions, variety, ripe-ness, maturity, and handling can be found (Martínez et al.2007). The total soluble solid content and acidity increasesduring ripening. The fat, ash, and protein contents are gener-ally higher in red pepper than in green peppers (see Table 1).In the case of Arnoia peppers (C. annuum L. var. annuum cv.Arnoia), potassium was the most abundant mineral both ingreen (2.80 g/100 g) and red peppers (251 g/100 g)(Martínez et al. 2007). Moreover, the quantity and composi-tion of bioactive compounds found in Capsicum spp. dependsfrom the extraction method and solvent used in its extraction(see Table 7). This results in a difficult accurately measure ofbioactive metabolites in different Capsicum varieties.

Thus, besides being used fresh or uncooked in many diets,fruits are subjected to several industrial transformations toconvert them to preserves, condiments, species, etc. Much ofthe nutraceutical value resides in their low calorie content andhigh antioxidant levels, especially ascorbic acid (vitamin C)and β-carotene (provitamin A). In fact, fruits are one of theagricultural products with the highest ascorbate content. Onehundred grams of Capsicum fruits provide approximately25 % of the recommended daily amount of vitamin A, and50 g of fresh fruit is enough to exceed the daily recommenda-tion for vitamin C (Palma et al. 2015; Guil-Guerrero et al.2006). Moreover, in the case of Jalapeño (C. annuum L.),the profiles of antioxidant capacity were completely differentfor green and red peppers and were more abundant in redfruits. Chlorophyll a (67.71 μg/g) and free all-trans-lutein(4.39 μg/g) were the major pigments in raw green peppers,whereas free all-trans-capsanthin (37.60 μg/g) was the mostabundant in raw red pepper. The heat treatment generated atleast 12 compounds, mainly pheophytins and cis isomers ofcarotenoids and reduced the fruits antioxidant capacity(Cervantes-Paz et al. 2012).

Capsicum fruits exhibit a high antioxidant activity due to awide variety of compounds such as phenols, flavonoids,capsaicinoids, and carotenoids. A strong correlation betweenthe presence of bioactive compounds in Capsicum spp. andtheir antioxidant activity evaluated by means of DPPH

radical-scavenging (Bae et al. 2012a, b), and ABTS (Xinet al. 2014) has been found in some studies, but not in others(Rahiman et al. 2013). This strong antioxidant activity playsan important role in the prevention of cardiovascular diseases,cancer, and neurological disorders. However, the content andbiodisponibility depend on the fruit ripening stage and thepost-harvesting process, so freezing and boiling process neg-atively influenced the content of these active compounds(Loizzo et al. 2015).

Water extracts from hot fruits of C. annuum L. (Tepin) andC. chinense (Habanero) have prevented ferrous lipid peroxi-dation in rat’s brain. However, Tepin fruits are more potentinhibitors than Habanero fruits; meanwhile, unripe Tepinshowed the highest protective ability due to its higher totalphenolic content and ferrous chelating activity; furthermore,while ripening decreased the antioxidant properties in Tepin, itshowed a reversed effect in Habanero increasing its antioxi-dant capacity (Oboh et al. 2007). The consumption ofC. frutensces (cayenne pepper) and their mixtures with garlicand ginger may help to modulate oxidative stress caused byhypercholesterolemia in rats (Otunola et al. 2010). Hot peppersuch as Jalapeño, Scotch Bonnet, and Bhut Jolokia showedidentical non-pungent water-soluble components similar tosweet peppers, regardless of their pungency level. This isprobably due to the biosynthetic origin of most of these com-pounds in the chloroplast. Moreover, hot chili contained otheranti-inflammatory compounds than capsaicinoids (Liu et al.2009).

Antioxidant Activity

Antioxidant Compounds in Capsicum spp.

As previously mentioned above, Capsicum spp. are known tobe a good source of different phytochemicals including vita-mins A and C, phenolic compounds, flavonoids and caroten-oids, among others. More than 125 volatile compounds havebeen identified in fresh and processed Capsicum fruits, al-though the significance of these compounds for the aroma isnot yet well known (El-Ghorab et al. 2013). Hot Capsicumvarieties are the only plants that are able to producecapsaicinoids, responsible for their characteristic hot taste.The concentration of these compounds depends on cultivar,maturity stage, growing conditions, and postharvest manipu-lation. In this way, the concentration of phenols varied from745 to 1032 mg/100 g, flavonoids from 201 to 389 mg/100 g,and ascorbic acid from 694 to 2153 mg/100 g in somejalapeño and Serrano cultivars (Alvarez-Parrilla et al. 2011).Some hot C. annuum L. (Jalapeño and Serrano), both freshand processed, are good sources of antioxidants such as phe-nolics and ascorbic acid (Alvarez-Parrilla et al. 2011).Moreover, these two varieties showed a lipid-protective effect

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mainly due to the presence of the phenolic compounds(Alvarez-Parrilla et al. 2012).

Capsaicinoids which consist on capsaicin, dihydrocapsaicin,nordihydrocapsaicin, homodihydrocapsaicin, and homocap

Table 7 Bioactive products from Capsicum spp. extracted with different solvents and extraction methods

Solvent used Extraction methods Bioactive compound extracted Reference

None Cold pressing Capsaicinoids Yılmaz et al. 2015

Acetone +Ethanol/water (9:1)

Fractioned extraction CapsaicinoidsCarotenoids

Amaya-Guerra et al. 1997

Ethanol/Water Fractioned extraction CarotenoidsCapsaicinoids

Santamaría et al. 2000; Dong et al. 2014

Acetonitrile HPLC Capsaicin and Dihydrocapsaicin Al Othman et al. 2011

Isopropanol + methanol/water HPLC Carotenoids Daood et al. 2002

Isopropanol + n-hexane HPLC Tocopherols Abbeddou et al. 2013

Acetone Maceration Carotenoids Hornero-Méndez and Mínguez-Mosquera2001

Acetonitrile Maceration Capsaicinoids Chinn et al. 2011

Comestible and medicinal oil Maceration Capsaicionoids Amruthraj 2014

Ethanol Maceration Capsaicinoids De Aguiar et al. 2014

Ethanol/Water Maceration Capsaicinoids Gao et al. 2005

Isopropanol Maceration Capsaicinoids Dorantes et al. 2000

n-Hexane Maceration Capsaicinoids Fernández-Ronco et al. 2013; Richinset al. 2010

Olive oil Maceration Capsaicinoids Paduano et al. 2014

Refined vegetable oil Maceration Carotenoids Guadarrama-Lezama et al. 2012

Water Maceration Capsaicinoids Pino et al. 2007

Acetone MAE Capsaicinoids Williams et al. 2004

Ethanol MAE Capsaicinoids Barbero et al. 2006

Methanol PLE Capsaicinoids Liu et al. 2014; Barbero et al. 2006

Water PHWE Capsaicinoids Bajer et al. 2015

CO2 SFE CarotenoidsCapsaicinoids

Perva-Uzunalić et al. 2004

CO2 + dimethyl ether SFE Carotenoids Catchpole et al. 2003

CO2 SFE Carotenoids Barros et al. 2015

CO2 SFE Capsaicinoids Kwon et al. 2011; Peusch et al. 1997;Sato et al. 1993; Duarte et al. 2004;de Aguiar et al. 2013; de Aguiaret al. 2014

CO2 + ethanol SFE CarotenoidsFree fatty acids

Ambrogi et al. 2002; Li et al. 2011

Propane SFE CarotenoidsTocopherolsFree fatty acids

Gnayfeed et al. 2001; Romo-Hualdeet al. 2012

CO2 SFE + UAE CapsaicinoidsPhenols

Santos et al. 2015

n-Hexane Soxhlet Capsaicinoids Fernández-Ronco et al. 2013; Richinset al. 2010

Ethanol UAE Capsaicinoids Boonkird et al. 2008

Hexane–methanol UAE Carotenoids Fernández-Ronco et al. 2013

Methanol UAE Capsaicinoids Barbero et al. 2014Sganzerla et al. 2014

MAEmicrowave-assisted extraction, PLE pressurized liquid extraction, PHWE pressurized hot water extraction, SFE supercritical fluid extraction,UAEultrasound-assisted extraction

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saicin can exert multiple pharmacological and physiological ef-fects. They are biosynthesized by condensation of fatty acids andvanillyllamine, and the placenta of the fruit is the major site fortheir biosynthesis. Capsaicinoids are stable in both polar andnonpolar solvents and produce the sensation of burning in thebody when comes into contact (Luo et al. 2011). In plants, cap-saicin purportedly plays a role in preventing microbial infectionsand suppressing unsuitable infestations as well as being a deter-rent to depredators (Singletary 2011). In human health, it hasdemonstrated benefits as a topical pharmaceutical to mitigatepain and other neurological conditions. However, the capacityof dietary capsaicin to manage gastrointestinal distress is unclear,due to the lack of understanding its apparent contradictory ac-tions within various segments of the gastrointestinal tract.Capsaicin’s pungency has limited its use in clinical trials to sup-port its biological activity (Reyes-Escogido et al. 2011). In thelast years, a lot of data has been published about linking capsaicinand hotCapsicum spp. to improve weight loss and weight main-tenance as a promising alternative, as well as leasing glucoseintolerance and insulin resistance. (Sharma et al. 2015;Singletary 2011). Consumed worldwide, the capsaicin presentin hot Capsicum spp. has a long story of controversy aboutwhether its safety. However, the US Food and DrugAdministration considered that theCapsicum fruits are complete-ly safe, but not pure capsaicin that it is considered not totally safe.High consumptions of hot fruits may be a risk factor for gastriccancer, but this is not entirely demonstrated (Bode and Dong2011).

Capsinoids are less pungent than its analogs ofcapsaicinoids found in some sweet cultivars of Capsicumspp. They consist mainly on capsiate, dihydrocapsiate, andnordihydrocapsiate, presenting a similar structure ascapsaicinoids. In the last years, capsinoids have been relatedwith a strong thermogenic activity and visceral fat burningand also have been successfully used to control weight man-agement without the inconvenient of capsaicinoidsconsumption.

The health benefits of non-pungent components fromCapsicum spp. such as carotenoids have been poorly ex-plored. Carotenoids are responsible for the yellow-orange-red color of Capsicum spp. and are composed by more than50 identified structures being the most important β-carotene,α-carotene, capsanthin, capsorubin, crytpocapsin, α-cryptoxanthin, β-cryptoxanthin, lutein, antheraxanthin,violxanthin, and zeaxanthin (Giuffrida et al. 2013). These pig-ments are commonly found in photosynthetic plants, algae,and microorganisms and play an important role in protectingtissues from light and oxygen.β-Carotene,α-carotene, andβ-cryptoxanthin are important sources of provitamin A.Carotenoids bearing a k-ring as end group have been shownto have strong reactive oxygen scavenging potential and aninverse relationship between human chronic disease incidentsand intake of carotenoids has been found. Other health

benefits such as cancer prevention and eye protection havealso been described (Arimboor et al. 2015; Abdel-Aal et al.2013).

Capsanthin, which is the major carotenoid present inCapsicum spp., is present in an acylated form with fatty acids.This carotenoid does not possess provitamin A activity but hasbeen shown to be effective as a free radical scavenger. It hasbeen proved that it has a plasma HDL cholesterol raising ef-fect in plasma (Aizawa and Inakuma 2009).

Phenolic compounds are found in considerable quantitiesin many fruits and vegetables and thus form an integral part ofthe human diet. Their consumption is related with reduced riskof cardiovascular diseases and certain types of cancer. InC. baccatum L. var. pendulum, the antioxidant activity waspositively correlated with the amount of phenolics found ineach sample (Kappel et al. 2008). In C. annuum L. cultivars,the phenolic content depended strongly on the cultivar, matu-rity stage, part of the fruit analyzed, and the drying process,freeze drying being the most conservative method althoughthe loss of phenolics could be higher than 50 % (Materska2014).

As previously said, all Capsicum spp. are a rich source ofan extensive diversity of bioactive compounds with potentialhealth benefits. However, the bioactivity depends on the ma-turity stage, the specific variety of Capsicum spp., and theextraction method (Conforti et al. 2007; Gahungu et al.2011; Jang et al. 2008; Aliakbarlu et al. 2014).

Antioxidant Activity in Different Extracts

Capsicum spp. hydrophilic fraction represents the main con-tribution to the total antioxidant activity of the fruits and main-ly depends on the cultivar and maturity stage of the fruit.Although small differences in the lipophilic fraction have beenfound under different cropping systems (organic or conven-tional) and the growth medium (soil or soilless) showinghigher lipophilic antioxidant content under organic conditions(López et al. 2014). The composition of lipophilic compoundsand the phenol profile varies during the different stages ofripening in C. annuum fruits modifying the antioxidant activ-ity. The radical scavenging activity increases with increasingof phenolic content for green and red chili extracts. Red pep-per fruits had potent antioxidant property (~450 CI μg/mLDPPH) (Conforti et al. 2007).

In general, there is scarce information about bioactivity ofwater extracts of Capsicum spp. (Aliakbarlu et al. 2014).Antioxidant activity of chili pepper varies, depending on thevariety, maturity stage, and technique used to measure thiscapacity (Gahungu et al. 2011; Jang et al. 2008; Aliakbarluet al. 2014). However, some studies have reported that aque-ous extracts of pepper can protect tissues from oxidative stressand from lipid peroxidation from diverse causes (Oboh et al.2007; Otunola et al. 2010). This protective effect from the

68 Food Bioprocess Technol (2017) 10:51–76

aqueous extracts is attributed to the presence of antioxidants,especially ascorbic acid and polyphenols (Oboh et al. 2007).Moreover, the consumption of C. frustescens may help tomodulate oxidative stress caused by hyper-cholesterolemiain rats (Outunola et al. 2014). Volatile compounds in waterextracts of some Habanero chili pepper cultivars have beenidentified and showed that orange and brown varieties have ingeneral higher amounts of esters that enhance their flavor-relevant chemical composition (Pino et al. 2007). In high hotScotch bonnet chili pepper, at the red stage were identified 70volatile compounds (Gahungu et al. 2011). However, in greenchili pepper, only 12 volatile compounds were found as majoraroma compounds (Jang et al. 2008).

Barbero et al. 2008 compared different extraction solventsto recover capsaicinoids from hot Cayenne pepper and foundthat water, which is a very polar solvent, has a poor capacity ofextraction. This reduced effectiveness is accentuated in thecase of less polar capsaicinoids such as dihydrocapsaicin,homocapsaicin, and homodihydrocapsaicin in comparisonwith more polar capsaicinoids as nordihydrocapsaicin andcapsaicin. On the other hand, Bajer et al. 2015 compared theextraction of capsaicinoids from ten chili samples usingPHWE and Soxhlet extraction obtaining better extractionyields in the first case with lower times and no contaminantsolvents (Bajer et al. 2015). In addition, in vitro studies aboutwater extracts of macerated Capsicum spp. reported anantimetastasic activity against human breast cancer cells(Kim et al. 2014) as well as antiobesity activity (Back et al.2013). Moreover, water extracts of bell Pepper showed ACEinhibitory activity (Kwon et al. 2011). Water extract ofC. pubescens showed ability to inhibit lipid peroxidation inrat’s brain homogenates probably due to its phenol contentand reducing power (Oboh et al. 2007). However, the mostfrequently use of water extracts from hot chili peppers is as asource of natural pesticides and insecticides for agricultureand even to reduce some human wildlife conflicts (Parkerand Osborn 2006).

Olive oil aromatized withC. frutescens showed an increasein its content of all the isoforms of vitamin E (from 186.4 incontrol to 198.6mg/kg olive oil) increasing also the nutritionalvalue but the phenolic content decreased from 345 in controlto 336 mg caffeic acid/kg olive oil in hot chili pepper (Sousaet al. 2015). However, the effect of the addition of hot chilipepper in the antioxidant potential is still unclear. Caporasoet al. (2013) found a significant increase in the infused oliveoil. However, other authors observed that the antioxidant po-tential decreases with the addition of dried chili (Baiano et al.2009; Sousa et al. 2015). Equally, the content of phenoliccompounds has been also reported lower than in unflavoredolive oil (Sousa et al. 2015; Baiano et al. 2009). The infusiontime and the dried chili pepper concentration are crucial for theproduction of flavored oils. Hence, the content ofcapsaicinoids in chili flavored oil is maximum at 7 days, and

no significant increase was observed for longer infusion timesand antioxidant activity is reduced after the first week of stor-age (Caporaso et al. 2013). The volatile composition of theolive oil was influenced by the concentration of dried chilipepper added as an increase in hexanal related to oxidationprocesses. The presence of dried chili pepper in enriched oliveoils with antioxidant compounds also modified its volatilesprofile. However, capsaicinoids and aroma compounds wererapidly released within the first week of chili infusion(Caporaso et al. 2013). Other study showed that, in long timestorage periods, the presence of a hot chili oleoresin improvedthe oxidative stability of the extra virgin olive oil (Gambacortaet al. 2007).

In the case of several cultivars of C. annuum extractedby Soxhlet with different organic solvents (hexane, ethylacetate, acetone, methanol, and methanol/water), thehighest levels of capsaicinoids and carotenoids werefound with hexane but the maximum level of flavonoidswere found with methanol (Bae et al. 2012b). Moreover,methanol extracts from C. annuum L. seeds showedstrong anti-proliferative activity against MCF7, MKn45,and HCT116 tumor cells at a concentration of 500 μg/mL due to an increase in apoptosis (Jeon et al. 2012).C. chinense (Habanero) ethanol extracts showed highamounts of antioxidant properties since the extractcontained compounds such as carotenoids, vitamins, phe-nolics, and capsaicinoids (Castro-Concha et al. 2014).C. annuum and C. chinense cultivars showed differentlevels of antioxidant activity and reducing activity basedon their content of capsaicinoids, carotenoids, flavonoids,and phenolics. The maximum antioxidant activity wasshowed using hexane as solvent while the inhibition ofdeoxyribose degradation was higher in methanol (Baeet al. 2012b). The extraction with acetonitrile of aC. fruscences variety presented more phenolics in com-parison when hexane was used as extraction solventresulting in a higher antioxidant activity (Nascimentoet al. 2014). In fact, polar aprotic solvents such as aceto-nitrile and acetone were more efficient to extractcapsaicinoids than non-polar solvents (Amruthraj 2014).Similarly, ethanolic extracts showed high flavonoids andphenol contents (Rahiman et al. 2013), but in some cases,low DPPH activity was found (Rahiman et al. 2013).Ethanolic and butanolic extracts from C. baccatum

contained potential antioxidant and anti-inflammatorycompounds which were tested against oxidative andinflammation-related pathological processes. The appro-priate use of solvents could be potentially usefulto measure and to extract the maximum antioxidantand nutritional value of Capsicum (Bae et al. 2012b).C. chacoense Hunz contained similar amounts of capsa-icin in comparison with C. baccatum and C. annuum L.Moreover, dichloroethane and ethanol extracts of

Food Bioprocess Technol (2017) 10:51–76 69

C. chacoense and C. baccatum L. elicited inhibition onthe araquidonic acid pathway in ear edema and could beused as well in human nutrition as phyto-preventives(López et al. 2014).

Seed extracts from C. annuum L. showed relatively lowantioxidant activity and polyphenolic content, but exertedhigh antiproliferative effects on tumor cells, even at low con-centrations (Jeon et al. 2012). Finally, Capsicum spp. oleo-resins have exhibited superoxide anion radical scavenging ac-tivity as well as antiproliferative activity inMCF7, HT-29, andHeLa cell lines (Šaponjac et al. 2014).

Antimicrobial Activity

The microbial safety of foods is one of the major concerns toconsumers, regulatory agencies, and food industries. Manyfood preservation strategies have been used traditionally forthe control of microbial spoilage in foods, but the contamina-tion is still an issue when is not adequately performed.Although many synthetic antimicrobials are approved in mostcountries, the recent trends are for the promotion and use ofnatural preservatives, which are safe, effective, and sensoryacceptable. Plants contain innumerable constituents and arevaluable sources of new and biologically active moleculeswith antimicrobial properties against a widespread variety ofbacteria, yeast, and molds. However, the variations in qualityand quantity of their bioactive compounds are the major dis-advantage in their use in foods. Further, phytochemical com-pounds added to foods may be lost by various processingtechniques such as high pressures and temperatures (Singhet al. 2014).

The antimicrobial activity in Capsicum spp. seems to bestrongly related with the presence of natural antioxidants suchas carotenoids than with the pungent components of the fruits.For that, some studies as Dorantes et al. (2000) found thatsweet red pepper (C. annuum L.) had more antimicrobial ac-tivity against common food pathogens than hot peppers suchas Jalapeño.

Fresh and heated water extracts of different Capsicum spp.have exhibited different degrees of inhibition against Bacilluscereus, Bacillus subtilis, Clostridium sporogenes, Clostridium

tetani, and Streptococcus pyogenes (Cichewicz and Thorpe1996). Moreover, a little activity against B. cereus, Listeriamonocytogenes, and Escherichia coli at high doses (150 mg/ml) has been found in Iranian red chili (C. annuum L.)(Aliakbarlu et al. 2014). The crude juice of C. frutenscenshas shown higher antimicrobial activity against E. coli,Salmonella typhi, and B. subtilis than organic solvent extractsas petroleum-ether, chloroform, isopropanol, and ethanol(Abdou Bouba et al. 2012). However, n-hexane and chloro-form extracts from C. frutescens L. showed inhibitory activityagainst Pseudomonas aeroginosa, Klebsilla pneumonia,Staphylococcus aureus, and Candida albicans (Gurnani

et al. 2015). Other research showed that water extracts ofcayenne pepper inhibited the presence of Enterobacter

aerogenes and L. monocytogenes (Kumral and Sahin 2003).In the case o C. baccatum L. var. pendulum, the ethanol

extract did not show any antimicrobial activity and the onlyantifungal activity was found in immature seeds ethanol ex-tracts (Kappel et al. 2008).

The antimicrobial activity of capsaicin microcapsules oncommon microorganisms for food preservation such asBacillus cinerea and Aspergillus niger have been also investi-gated. The factors affecting the antimicrobial effects, includingthe microcapsule concentrations, pH values, and release behav-ior have also been examined. The shelf life of low pH foods(acidic) tends to be better than high pH foods (alkaline) wheremicroorganism can better growth. The optimum antimicrobialeffect in Capsicum oleoresins was found at pH 5.0 in short-term storage foods (Xin et al. 2014). Supercritical CO2 andethanol extracts from C. annuum and C. frutenscens showedantibacterial effect against Streptococcus sobrinus andS. salivarius. Therefore, there are promising materials for newantiseptical agents for oral care products (Pilna et al. 2015).

Conclusions

Oleoresins are natural products that consist of complexmixtures mainly of lipophilic molecules. C. oleoresinshave been used for several applications in pharmaceutical,cosmetic, agricultural, and food industries mainly for itsensorial attributes (color and pungency). Moreover,Capsicum spp. oleoresins are rich in bioactive compoundswith antimicrobial and antioxidant activities such as carot-enoids and capsaicinoids that can be used as a natural ad-ditive. However, Capsicum spp. oleoresins have been tra-ditionally extracted by organic solvents that potentiallyrisk both for the environment and human health.Research shown here display the increase of green extrac-tion technologies at the laboratory level. Nonetheless, mostof these non-conventional extraction technologies requirehigh production costs, making it unfeasible for its devel-opment in emergent countries that are the main Capsicum

spp. producers. In this regard, non-conventional extractionwith vegetable oils becomes a good green and cost-efficient alternative to extracting Capsicum compounds.Finally, there is a strong relation between the extractiontechniques and the compounds extracted in the case ofCapsicum spp. oleoresins due to the different polarity ofthe extraction solvents. Differences in the polarity of thebioactive compounds to be extracted allow the productionof Bintelligent^ oleoresins rich in the selected bioactivecompound. Further research should be carried out to un-derstand this matter.

70 Food Bioprocess Technol (2017) 10:51–76

Acknowledgments This research was partially supported by ConsejoNacional de Ciencia y Tecnología (Conacyt) through a postdoctoral grantwith the agreement number 290807-UV.

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