Critical Reviews in Biotechnology, 28:285–296, 2008

Copyright
c Informa UK Ltd.

ISSN: 0738-8551 print / 1549-7801 online
DOI: 10.1080/07388550802368895

Processing of Apple Pomace for Bioactive Molecules

Shashi Bhushan, Kalpana Kalia, Madhu Sharma, Bikram Singh,
and P. S. Ahuja
Institute of Himalayan Bioresource Technology (CSIR), Palampur, Himachal Pradesh, India
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The growth of horticulture industries worldwide has generated huge quantities of fruit wastes
(25%–40% of the total fruits processed). These residues are generally a good source of carbohy-
drates, especially cell wall polysaccharides and other functionally important bioactive molecules
such as proteins, vitamins, minerals and natural antioxidants. “Apple pomace” is a left-over
solid biomass with a high moisture content, obtained as a by-product during the processing of
apple fruits for juice, cider or wine preparation. Owing to the high carbohydrate content, apple
pomace is used as a substrate in a number of microbial processes for the production of organic
acids, enzymes, single cell protein, ethanol, low alcoholic drinks and pigments. Recent research
trends reveal that there is an increase in the utilization of apple pomace as a food processing
residue for the extraction of value added products such as dietary fibre, protein, natural an-
tioxidants, biopolymers, pigments and compounds with unique properties. However, the central
dogma is still the stability, safety and economic feasibility of the process(s)/product(s) developed.
This review is mainly focused on assessing recent research developments in extraction, isolation
and characterization of bioactive molecules from apple pomace, along with their commercial
utilization, in food fortification.
For personal use only.

Keywords apple pomace, biocatalyst, dietary fibre, natural antioxidant, pectin, pigment

INTRODUCTION worldwide (Table 1). It has high moisture content (70%–75%)

The research and development efforts on value addition and
efficient utilization of nutritionally rich agro-industrial residues
such as whey, sugar beet pulp, cassava bagasse, apple po-
mace, citrus waste, coffee pulp/husk, etc are gaining momen-
tum around the world. The overall strategy for the utilization
of such food processing residues is generally built around fer-
mentative or non-fermentative product development. In the case
of fermentative utilization, the residues are used as microbial
substrates for the development of a variety of value added prod-
ucts (Kennedy, 1994; Joshi and Pandey, 1999; Pandey et al.,
2000; Lynd et al., 2005; Venus and Richter, 2007; Vendrus-
colo et al., 2008). In non-fermentative concepts, agro-industrial
residues are processed for the extraction of bioactive molecules
(Larrauri, 1999; Laufenberg et al., 2003; Schieber et al., 2003;
Nawirska and Kwaúniewska, 2005).
“Apple pomace” is a left-over solid residue (25%–30% of
the total processed fruits) obtained after the extraction of ap-
ple juice. Several million tonnes of apple pomace are generated
Address correspondence to Shashi Bhushan, Institute of Himalayan
Bioresource Technology (CSIR), Post Bag-6, Palampur-176 061,
Himachal Pradesh, India. Tel.: +911 894233339387; Fax: +911
894230433; E-mail: sbhushan@ihbt.res.in
and biodegradable organic load (high BOD and COD values).
High moisture content makes apple pomace bulky and suscep-
tible to microbial decomposition. Huge piles of apple pomace,
outside or near the industrial unit, not only violate the pollution
control norms and industrial safety issues, but also lead to public
health hazards. Owing to high transportation costs (being bulky)
and the generation of foul smells (fast biodegradation by natural
flora), direct dumping is also a problem. Earlier, sun energy was
used for drying apple pomace in the open to reduce the bulk.
This method makes apple pomace darker (enzymatic or oxida-
tive browning) and unfit for value addition, especially for human
food fortification. In addition to sun drying, a number of meth-
ods have been reported for reducing the bulk of apple pomace
and to preserve it for further utilization (Fenton and Kennedy,
1998; Constenla et al., 2002; Sun et al., 2007). The selection of
a particular method for pomace drying depends upon its energy
cost, change in nutritional profile and intended purpose.
Intrinsic to apple fruit nutrients, pomace also contains numer-
ous phytochemicals in the form of simple sugars, pectin, dietary
fibres and natural antioxidants. In the past, apple pomace dried
for use as animal feed, fuel for boilers or added to soil as a con-
ditioner (Singh and Narang, 1992; Takahashi and Mori, 2006).
Since 1980, it has been extensively studied as a substrate for
microbial growth and used in the production of value added

285
 286 S. BHUSHAN ET AL.

TABLE 1 Joshi, 2006; Attri and Joshi, 2006; Gullón et al., 2007). Ven-
World apple pomace generation druscolo et al. (2008) recently reviewed in detail the research
and development efforts currently underway on apple pomace
Quantity throughout the world for fermentative product development.
Country (thousands of tonnes) Reference Since 1985, a continuous increase was observed in the num-
Spain 20 Gullón et al., 2007 ber of publications in peer reviewed journals per year (Figure 1a)
Germany 250 Endreß, 2000 dealing with apple pomace utilization. In this period, R & D ini-
New Zealand 20 Lu and Foo, 1998 tiatives were more focused, on apple pomace conversion into
India 3–5 HPMC, 2007 value added product development (Joshi et al., 1996; Masoodi
Brazil 13.75 Villas-Bôas et al., 2003 et al., 2002; Sudha et al., 2007), or on extracting the bioactive
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Iran 97 Pirmohammadi et al., 2006 constituents for food fortification or food enrichment (Fig. 1b).
Japan 160 Takahashi and Mori, 2006 Efforts were also made to understand the addition of apple po-
United States 27 Roberts et al., 2004 mace on the nutritional profile of prepared food products and
on regulatory mechanisms, as evident from the percentage of
publications in the area of nutrition and dietetics as well as bio-
chemistry and molecular biology (Masoodi et al., 2002; Zhang
products such as organic acids, enzymes, single cell proteins, et al., 2003; Sembries et al., 2003; Devarajan et al., 2004;
low alcoholic drinks, ethanol, biogas, pigment and baker’s yeast Lantto et al., 2006; Sehm et al., 2006; Fridrich et al., 2007;
(Hang et al., 1982; Sharma, 1983; Kalia et al., 1992; Wiacek- Gutzwiller et al., 2007). The fact is also supported by the utiliza-
Zychlinska, et al. 1994; Hang and Woodams, 1995; Shojaosadati tion and disposal hierarchy of food processing residuals (Brandt
and Babaeipour, 2002; Seyis and Aksoz, 2005; Bhushan and and Martin, 1994), according to which recovery of human
For personal use only.

FIG. 1. R & D publications on apple pomace: in relation to (a) year-wise and (b) subject-wise percentage (Source: Web of Sciences,
2008).
 PROCESSING OF APPLE POMACE FOR BIOACTIVE MOLECULES
consumption products from agricultural residues has been pri-
oritized at the second position of the management strategies.
Besides fermentative product development, since the 1920s, ef-
forts have continuously focused on the isolation of bioactive
molecules from apple pomace, with the execution of a pectin
extraction industrial unit in the USA and Europe. In addition
to pectin, apple pomace is also used for extraction of dietary
fibre, xyloglucan, natural antioxidant and aromatic compounds
(Carson et al., 1994; Almosnino et al., 1996; Barwal and Kalia,
1997; Watt et al., 1999; Lu and Foo, 1997; Schieber et al., 2003;
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Canteri-Schemin et al., 2005; Figuerola et al., 2005; Royer et al.,
2006; Guyot et al., 2007). Apple pomace has also been consid-
ered for new applications such as textile dye removal (Robinson
et al., 2002), press aid (Roberts et al., 2004), as a heavy metal
absorbent (Nawirska, 2005) and as a protein stabilizer (Lantto
et al., 2006). Thus, value addition of such residues will not only
provide the means to industrial economic stability but will also
help in reduction of environmental pollution, judicious man-
agement of natural bioresources and availability of nutrition-
ally enriched food products, as well as industrially important
biomolecules.
APPLE PROCESSING AND ITS BY-PRODUCTS
The world production of apples in 2006–2007 was 46.1 mil-
For personal use only.
lion tonnes, with more than a 50% (24.5 million tonnes) con-
tribution from China, followed by the USA. China is slowly
capturing the world apple market in fresh, as well as pro-
cessed, products. India is far behind and holds ninth posi-
tion but is among the top 25 apple producing countries in the
world with a total production of 1.3 million tonnes, (MFPI,
2006). Out of the total production, about 71% of the fruit is
marketed as being fresh, while 25%–30% of the fruit is pro-
cessed into juice, cider, and frozen and dried processed products
(Figure 2).
Apple juice concentrate (AJC) is the major processed product
throughout the entire world (64% of the total of all processed
FIG. 2. World apple production and processing status.
287
apple products) and is prepared either from specific processing
varieties or low grade and culled fruits that are otherwise un-
suitable for the fresh market. Global apple juice production was
estimated at 1.4 million metric tonnes during 2004–2005, with
China’s contribution alone being 600,000 tonnes. India produces
4500 tonnes per annum of AJC and this is equivalent to about
0.64% of the total world production. The total recovery of the
juice in industrial production process (Figure 3) is about 70%–
75%, generating 25%–30% apple pomace and 5%–11% sludge
(liquid waste obtained after clarification and sedimentation with
bentonite). Apple pomace consists mainly of apple skin/flesh
(95%), seeds (2%–4%) and stems (1%). In India, around 3000–
5000 tonnes of apple pomace are generated annually, with 2000–
3000 tonnes in Himachal Pradesh alone. Sludge contains apple
juice (5–6◦ Brix TSS) and bentonite particles (added during clar-
ification of the juice), and there seems to be a direct loss of about
10% juice, which can be recovered easily by adoption of efficient
processing techniques.
NUTRITIONAL PROFILE OF APPLE POMACE
The apple fruit is highly nutritious and contains carbohy-
drates, proteins, minerals and natural antioxidants. These fruits
are invaluable in terms of:
• Elimination of certain harmful substances from the
body.
• Prevention of decomposition of protein matter in the
alimentary canal.
• The malic acid being beneficial to the bowels, liver and
brain.
• Help in prevention of anemia, constipation, stomach
disorders, hypertension, rheumatic afflictions, etc.
As a fresh fruit, apple pomace contains significant amounts
of phytochemicals such as carbohydrates, proteins, vitamins and
minerals. The nutritional profile (Table 2) of dried apple pomace
reveals a high content of carbohydrates with a fermentable sugar
 288 S. BHUSHAN ET AL.
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For personal use only.
FIG. 3. Industrial processing of apple juice concentrate (AJC).
content of up to 50% (Hang and Walter, 1989; Bhushan, 2002;
Gullón et al., 2007). The apple pomace carbohydrates mainly
consist of galacturonic acid (49%–64%), arabinose (14%–23%)
and galactose (6%–15%), with minor amounts of rhamnose, xy-
lose and glucose (Mehrländer et al., 2002). Besides carbohy-
drates, apple pomace is rich in proteins, vitamins (A and C)
and minerals (P, K, Mn, Ca, Mg and Fe) (Hang and Walter,
1989; Johr et al., 1960; Kennedy, 1994). It is a good source of
natural antioxidants (catechins, procyanidins, caffeic acid, phlo-
ridzin, phloretin glycosides, quercetin glycosides, chlorogenic
acid, etc). Apple pomace, including seeds, contains polypheno-
lics (Table 3) with the strong antioxidant activity of quercetin
glycosides, phloridzin and its oxidative products (Lu and Foo,
2000; Schieber et al., 2002; Guyot et al., 2007). The seed con-
tains about 80% of fatty acids, the majority being linoleic acid
and oleic acid.
PROCESSING OF APPLE POMACE FOR BIOACTIVE
MOLECULES
Dietary Fibre
“Dietary fibres” are the plant cell wall compounds (non-starch
polysaccharides), which are resistant to hydrolysis by digestive
enzymes in humans. They are intrinsic and intact in plants and
consist of mainly cellulose, hemicelluloses, pectic substances
and lignins (Trowell, 1974). Diets with inadequate dietary fibre
content are generally associated with constipation, diverticu-
losis, cardiovascular disease and cancer (Trowell, 1985). Di-
etary fibres are known to exert a number of protective effects
on cardiovascular diseases, colorectal cancer, obesity and di-
abetes (Salmeron et al., 1997; Ellegárd et al., 2000). Fibres
bind excess hydrochloric acid, cholesterol and gastric juices,
increase the faecal bulk and stimulate intestinal peristalsis (Jenk-
ins et al., 1998; Nawirska and Kwasniewúka, 2005). The fruit-
and vegetable-based fibres are known to have better nutritional
quality than cereal formulations, because of the presence of sig-
nificant amounts of associated active compounds (flavonoids,
carotenoids, higher fibre content) in balanced composition (sol-
uble/insoluble dietary fibre ratio, water- and fat-holding capac-
ities, lower energy value and phytic acid content).
Apple pomace contains a significant amount of non-starch
polysaccharides (35%–60% dietary fibre), with a high amount
of insoluble fibre (36.5%) as well as soluble fibre (14.6%) (Chen
et al., 1988; Gallaher and Schneeman, 2001; Villas-Bôas et al.,
2003; Sudha et al., 2007). The main constituents of these dietary
fibres are pectins (5.50%–11.70%), cellulose (7.20%–43.60%),
hemicelluloses (4.26%–24.40%), lignins (15.30%–23.50%) and
gums. Cellulose, pectin and lignin are water insoluble, while
galacturonic acid and hemicelluloses are water soluble. A num-
ber of fibre enriched bakery products were prepared by adding
dried apple pomace powder on a wheat flour replacement basis.
 PROCESSING OF APPLE POMACE FOR BIOACTIVE MOLECULES 289

TABLE 2
Proximate composition of apple pomace

Composition Composition
Constituents (dry weight basis) Constituents (dry weight basis)

Moisture (%) 3.90–10.80 Alcohol-soluble fraction of carbohydrate

Protein (%) 2.94–5.67 Saccharose (%) 3.80–5.80
Total carbohydrate (%) 48.0–62.0 Glucose (%) 19.50–19.70
Fibre (%) 4.70–51.10 Fructose (%) 48.30
Insoluble fibre 36.50 Xylose, mannose and galactose (%) 1.20–4.40
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Soluble fibre 14.60 L-malic acid (%) 2.60–3.20

Fat (ether extract, %) 1.20–3.90 Arabinose and rhamnose (%) 7.90–6.0
Pectin (%) 3.50–14.32 Glucooligosaccharides (%) 3.40–3.80
Ash (%) 0.50–6.10 Xylooligosaccharides (%) 3.0–3.70
Minerals Arabinooligosaccharides (%) 0.20–0.40
Phosphorus (%) 0.07–0.076 Uronic acid (%) 2.70–3.40
Potassium (%) 0.43–0.95 Alcohol-insoluble fraction of carbohydrate
Calcium (%) 0.06–0.10 Glucan (%) 41.90–42.90
Sodium (%) 0.20 Starch (%) 14.40–17.10
Magnesium (%) 0.02–0.36 Cellulose (%) 7.20–43.60
Copper (mg/kg) 1.10 Polysaccharides of xylose, mannose and galactose (%) 13.0–13.90
Zinc (mg/kg) 15.00 Polysaccharide of arabinose and rhamnose (%) 8.10–9.0
Manganese (mg/kg) 3.96–9.00 Acid detergent lignin (%) 15.20–20.40
Iron (mg/kg) 31.80–38.30 Uronic acid (%) 15.3
For personal use only.

Sources: Chen et al., 1988; Hang and Walter, 1989; Kennedy, 1994; Joshi and Sandhu, 1994; Masoodi and Chauhan, 1998; Bhushan, 2002;
Constenla et al., 2002; Schieber et al., 2003; Marcon et al., 2005; Virk and Sogi, 2004; Nawirska and Kwaúniewska, 2005; Sudha et al.,
2007; Gullón et al., 2007)

Chen et al. (1988) investigated the effect of fibre source and
fibre concentration on physical and baking properties of bread,
cookies and muffins. They used commercially available fibres
of apple (spray dried), wheat bran and oat bran at 0%, 4%, 8%
and 12% concentration with uniform particle size (130 µm).
The results showed that apple fibres had a higher content of
dietary fibre (61.90%), lipids (2.45%), and lower amounts of
proteins (7.25%) and ash (1.27%), than other fibre sources. Irre-
spective of fibre source, a significant increase in water absorp-
tion, product weight and dough development time was observed
with the increase in fibre concentration. A chemical analysis of
the finished product showed that the bakery products prepared
by adding apple fibres had a higher dietary fibre content than
other sources. Hence, the authors have advocated the addition
of apple fibres (up to 4%) for food enrichment without compro-
mising product quality and acceptance by consumers. In another
study, cakes were prepared with direct incorporation of apple po-
mace in the wheat flour and the baking properties were evaluated
(Masoodi et al., 2002). They used dried apple pomace powder
of different particle sizes (30 µm, 50 µm and 60 µm) with a
wheat flour replacement basis at 0%, 5%, 10% and 15% con-
centration. A significant increase in batter viscosity (82.6 poise,
113.6 poise and 307.6 poise) was observed with an increase in
pomace concentration (5%, 10% and 15% pomace) compared
to the control (64.6 poise) at uniform particle size (30 mesh).
The results revealed a significant increase in cake weight with
the increase in pomace concentration and particle size, which
caused the batter properties to deteriorate to an unacceptable
level. Cakes were also prepared by direct incorporation of finely
ground apple pomace (150 µm) in wheat flour at 0%, 10%, 20%
and 30% on a flour replacement basis (Sudha et al., 2007). The
rheological properties of the dough revealed an increase in wa-
ter absorption, resistance to extension and pasting temperature,
with the increase of apple pomace content in the dough. They
observed a significant increase in cake weight and texture, and
a decrease in cake volume with the increase in concentration of
apple pomace. Nutritional characteristics showed that products
prepared with apple pomace incorporation (25% apple pomace
blends) had significantly higher amounts of dietary fibre (14.2%)
as compared to the control (0.47%). It was inferred that apple
pomace not only improved the dietary fibre content of the baked
product, but also increased the phenolic content (3.15 mg g−1 ) at
a 25% addition over the control (2.07 mg g−1 ). However, these
findings still require validation on a product safety and stability
scale before recommendations for commercial food fortification
or nutritionally enriched product development can be made. It
is worth mentioning that there is still no accessible report on
apple pomace derived food products for human consumption on
a commercial scale. Hence there seems to be a wide scope in
this segment of food processing.
 290 S. BHUSHAN ET AL.

TABLE 3 delivery (Holst et al., 2006). The gelation properties of pectin

Total phenolic compounds identified in apple pomace fractions are a major factor in terms of its suitability for a particular appli-
and seeds cation and gelation these properties are known to be influenced
by the length of side-chain branches, the degree of acetylation
Apple pomace and the pattern of esterification (Constenla et al., 2002).
Catechin Quercetin 3-arabinofuranoside Apple pomace consists of approximately 10%–15% pectin
Epicatechin Quercetin 3-rhamnoside on a dry weight basis (Endreß, 2000; Wang et al., 2007). Pectin
p-Coumaric acid Quercetin-O-pentoside from dried apple pomace is usually extracted with diluted min-
p-Coumaroylquinic acid Sinapic acid-O-glucoside eral acids at elevated temperatures to solubilize the protopectin,
Caffeic acid-O-glucoside Apigenin* followed by alcohol precipitation (May, 1990). The dried apple
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Chlorogenic acid
Caffeoylquinic acid
Cyanidin 3-glucoside
Dicaffeoylquinic acid
Ferulic acid
3-Hydroxyphloridzin
Kaempferol-O-glucoside
Procyanidin B2
Phloridzin
Phloretin
Phloretin xyloglucoside
Quercetin
Quercetin 3-diglucoside
Cyanidin 3-glucoside
For personal use only.
Chrysoeriol*
Eriodictyol-hexoside*
Eriodictyol*
Hesperidin-O-pentoside*
Luteolin*
Luteolin-7-O-glucoside*
Luteolin-7-O-galactoside*
Naringenin*
Naringenin-7-O-rutinoside*
Naringenin-O-hexoside*
Naringenin-O-hexoside*
Naringenin-O-glucuronide*
Naringenin-7-O-neohesperidoside*
Naringenin-7-O-glucoside*
pomace is ground to powder (particle size 80 mesh) and extru-
sion is performed (by twin-screw extruder) at different speeds,
feed rates and moisture content (Hwang et al., 1998). Extrusion
has been found to exert a more solubilizing effect than the com-
mercial acid extraction process. However, it is effective only in
increasing the pectin yield with a lower degree of esterification,
which has limited industrial application. The authors have ob-
served a cohesive effect of pH of the buffer, extraction time and
solid:liquid ratio on pectin yield. The pomace obtained from
Granny Smith apples, on a laboratory scale, was dried at differ-
ent temperatures (60◦ C, 70◦ C, 80◦ C and 105◦ C) to evaluate the
effect on pectin characteristics, composition, colour and gelpoint
temperature (Constenla et al., 2002). Galacturonic acid content
seemed to be unaffected by the drying temperature; however, sig-

Quercetin 3-rutinoside Protocatechuic acid* nificant effects were noticed on pectin colour and the gelpoint
Quercetin 3-galactoside Quercetin-O-pento-hexoside* temperature (maximum at 80◦ C). Extremes of drying temper-
Quercetin 3-glucoside Rhamnetin* ature resulted in more brownish pectin colour, either owing to
Quercetin 3-xylanoside Rhamnetin 3-glucoside* enzymatic browning (at a lower drying temperature, 60◦ C) or
Quercetin-O-hexoside Salicylic acid* caramelization (at a higher drying temperature, 105◦ C). Canteri-
Quercetin Schemin et al. (2005) used nine apple varieties to study the effect
3-arabinopyranoside of variety, particle size and kind of acid used for extraction on
Apple seed pectin yield. The apple pomace flour with a particle size higher
Catechin Phloridzin than 600 µm showed a lower pectin yield (maximum between
Epicatechin Phloretin 106 µm and 250 µm) and a statistical difference was noticed
p-Coumaric acid Phloretin xyloglucoside among the varieties with respect to pectin yield. However, this
p-Coumaroylquinic acid Quercetin 3-galactoside has little relevance to industrial apple pomace processing, as
Chlorogenic acid Quercetin-3-O-glucoside there is always a combination of number of varieties. Among
Procyanidin B2 Quercetin-3-O-rhamnoside the acid used for pectin extraction, citric acid showed the highest
average yield (13.75%) and was reported to be more econom-
*Reported for first time in apple or apple pomace fraction. ical and safe. Wang et al. (2007) used a microwave-assisted
Sources: Lu and Foo, 1997, 2000; Schieber et al., 2002, 2003; Sánchez- extraction system, at a frequency of 2450 MHz, for extraction
Rabaneda et al., 2004; Guyot et al., 2007, Cetkovic et al., 2008. of pectin from dried apple pomace powder (particle size 0.6 ×
10−3 to 1.5 × 10−3 ) and the analytical data was verified using
Pectin response surface methodology. Microwave-assisted extraction
The utilization of apple pomace for pectin production is con- reduced the pectin extraction time considerably. The authors
sidered to be one of the most practical approaches. Fruit pectins concluded from the experimental results that “extraction time
have been employed as gelling agents, thickener texturizers, and buffer pH” and “pH and solid:liquid ratio” had significant
emulsifiers and stabilizers in food processing, and the cosmetics effects on pectin yield and on the degree of methylation. The op-
and pharmaceutical industries (Thakur et al., 1997). In the food timum predicted yield (0.158 g g−1 dried apple pomace powder)
industry, pectin is used in products such as jams and jellies; fruit was recorded with pH 1.01 and HCl buffer, 499.4 W microwave
preparation; bakery jellies; confectionery; yogurt and acidified power, 20.8 min extraction time and 0.069 solid:liquid ratio. Re-
milk drinks; beverages; and frozen foods. The non-toxicity and cently, the effect of extraction conditions on yield and purity of
biocompatibility of fruit pectin render its application as a car- apple pomace pectin by alcoholic precipitation was investigated
rier for drugs, e.g. for oral administration of colon-specific drug (Garna et al., 2007). The extraction was carried out using two
 PROCESSING OF APPLE POMACE FOR BIOACTIVE MOLECULES
different conditions of pH and process time by heating 50 g of
apple pomace in 1 L of acidic water, with continuous stirring
at 250 rev min−1 in a jacketed stainless steel reactor at a tem-
perature of 90◦ C. The low pH (1.5) with the longer extraction
period (3 h) resulted in a high yield of pectin (8.9%), while a
slight increment in pH (2.0) with a decrease in extraction time
(2 h) produced pectin with a high content of galacturonic acid.
The researchers were able to obtain pectin with a high degree of
methylation from both extraction conditions, but with a brown
hue that developed owing to the oxidation of phenolic com-
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pounds. Efforts were made to improve the colour of the apple
pomace pectin by bleaching it with an alkaline peroxide (Re-
nard et al., 1996). The process helped in improving the extracted
pectin colour, but led to a loss of polyphenolics and to a degra-
dation of the major part of the pectins, resulting in a lower pectin
yield. Later, Schieber et al. (2003) developed a method for the
combined recovery of pectin and polyphenolics. This process
helped the production of a light coloured pectin with the simul-
taneous recovery of phenolic compounds, which can be used as
a source of antioxidants. At present, fruit pectins with varying
properties are in great demand and hence there is a need for
processes for the development of products with tailored proper-
ties at an industrial level, and this is a great challenge for R&D
personnel.
For personal use only.
Xyloglucan
In addition to pectin, apple pomace is also a good source
of cellulose and hemicelluloses – xyloglucan mostly fuco-
galactoxyloglucans, representing up to 18% of apple cell wall
polysaccharides (Renard et al., 1995; Watt et al., 1999; Caili
et al., 2006). Cellulose is used for the production of various
industrial products (methylcellulose, hydroxypropylcellulose,
carboxymethylcellulose) having wide commercial applications.
However, the exploitation of xyloglucan extraction from apple
pomace is at an infant stage. Xyloglucans are not solubilized dur-
ing the hot acidic treatment employed for pectin extraction. Re-
nard et al. (1995) suspended depectinized apple pomace (1 g) in
100 mL alkali solution (pH 7.0) and centrifuged after incubation
for 15 min. They found that the increased alkali concentration
and extraction time resulted in high xyloglucan production.
Xyloglucan was extracted with 4 M KOH from a commercial
apple pomace source with and without pectinase pretreatment
(Watt et al., 1999). Different oligosaccharides were isolated from
the 1,4-β-D-glucanase hydrolysate and characterized using 13 C
NMR spectroscopy. Enzymatic treatment of the apple pomace
resulted in the production of lower molecular weight xyloglucan,
with a lower viscosity compared to non-enzymatic treatment. An
almost similar composition of neutral sugars was found in all
the purified xyloglucan fractions, except for a slight variation
owing to apple variety and type of pomace used for extraction.
The intrinsic viscosity of the purified xyloglucan was reported
to be 244 mL g−1 . The results were comparable to earlier re-
ports of Renard et al. (1995) (240 mL g−1 ), despite different
extraction procedures. In both studies, the yield of xyloglucan
291
was reported to be influenced by the liquid:solid ratio, time of
extraction, concentration of alkali, type of pomace and pro-
cess temperature. Furthermore, ultrasound-assisted extraction
(50 kHz frequency with an input power of 160 W) was observed
to reduce the time by a factor of three over the alkali extraction
method (Caili et al., 2006). The three independent variables,
i.e. liquid (34.4):solid (1) ratio, KOH concentration (3.3 M) and
extraction time (2.5 h) were optimized using response surface
methodology. The conversion of xyloglucan into compounds
such as thickening agents and texture modifiers or as a source
of biologically active oligosaccharides having medicinal prop-
erties, is possible. Therefore the extraction of xyloglucan from
apple pomace can provide a new means for the development of
high-value biomolecules.
Natural Antioxidants
A range of polyphenolics compounds have been isolated
from apple pomace, such as cinnamic acid and its deriva-
tives, epicatechin, epicatechin dimer (procyanidin B2), trimer,
tetramer and oligomer, quercetin-3-glycosides phloridzin and
3-hydroxyphloridzin (Table 3). The nature of procyanidin
present in apple pomace has been established by 13 C-NMR
spectroscopy, by the acid-catalyzed degradation reaction with
phloroglucinol and by electro-spray mass spectrometry (Foo and
Lu, 1999). Earlier, these authors had investigated Gala apple po-
mace as a potential source of polyphenols (Lu and Foo, 1997).
The fraction was obtained by extracting apple pomace powder
with 70% aqueous acetone. The extract was concentrated and
left-over residue defatted with hexane, concentrated and freeze
dried before separation on Sephadex LH-20 using methanol as
solvent. Out of the total polyphenols present (7.24 g kg−1 DM),
quercetin glycosides accounted for more than half (4.24 g kg−1
DM), followed by phloridzin and its oxidative products. The elu-
tion order of the seven quercetin glycosides from apple pomace
was established by Schieber et al. (2002) using HPLC stationary
phase with hydrophilic endcapping system. The separation was
achieved within 8 min for all compounds and the elution order
was found to be Q-3-O-rutinoside, Q-3-O-galactoside, Q-3-
O-glucoside, Q-3-O-xyloside, Q-3-O-arabino-pyranoside, Q-
3-O-arabino-furanoside and Q-3-O-rhamnoside. The predom-
inant quercetin glycosides detected were Q-3-O-galactoside,
Q-3-O-arabinosoide, Q-3-O-glucoside and Q-3-O-xyloside.
In the subsequent year, Schieber et al. (2003) reported a pro-
cess for the simultaneous recovery of pectin and polyphenols
from apple pomace. The acidic extract of apple pomace (pH 2.8,
TSS 3.1◦ Brix) preheated to 60◦ C was passed through an Amber-
lite XAD 16 HP resin- (1.77 L) containing glass column at a flow
rate of approximately 10 bed volumes per hour. Pectins present in
the extract easily pass through the resin and pectin-containing ef-
fluent was collected at the bottom. Pectin residues were removed
with distilled water until no pectin could be recovered by alco-
hol precipitation. The process resulted in a light coloured pectin
with a slight increase in the degree of esterification and galactur-
onic acid units. The adsorbed phenolic compounds were eluted
 292 S. BHUSHAN ET AL.

TABLE 4

Apple pomace (mg kg−1 DM)

Schieber et al., 2003

Apple seed (mg kg−1 DM)
Phenolic compounds Lu and Foo, 1997 Drum dried Lyophilisate Schieber et al., 2003

Catechin nd* nd 2,400 nd

Epicatechin 640 nd 9,300 9.6
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by North Carolina State University on 03/11/13

Caffeic acid 280 nd nd nd

3-Hydroxyphloridzin 270 nd nd nd
Chlorogenic acid nd nd 14,300 119.8
Phloretin-xyloglucoside 170 nd nd 47.4
Phloretin nd nd nd 6.3
Phloridzin 1,420 nd 11,400 1,915.0
Quercetin 3-galactoside 1,610 224.2 nd 12.9
Procyanidin B2 nd nd 9,300 17.0
Quercetin 3-glucoside 870 27.6 3,900 5.9
Quercetin 3-xyloside 530 114.4 1,800 nd
Quercetin 3-arabinoside 980 223.0 1,100 nd
Quercetin 3-rhamnoside 470 297.5 4,700 25.0
p-Coumaroylquinic acid nd nd 1,800 9.4
*nd = not detected
For personal use only.

with a two bed volume of methanol, and after solvent removal in
vacuo, residues were lyophilized at 0.075 mbar for 80 h. The to-
tal lyophilisate contained about 12% of phenolic compounds and
a fraction was characterized by HPLC using a stationary phase
with hydrophilic endcapping. The main phenolic compounds
detected were phloridzin, chlorogenic acid and quercetin-3-O-
glycosides, with appreciable amounts of catechin and procyani-
din. The authors also investigated the phenolic profile of wet
and three-stage drum-dried apple pomace. Industrial drying of
apple pomace has been reported to have a non-significant ef-
fect on the total yield or phenolic profile except for flavanols,
which were adversely affected. Sánchez-Rabaneda et al. (2004)
identified about 60 phenolic compounds in different apple po-
mace fractions (peel, core, seed, calyx, stem and soft tissues)
using LC/MS/MS and also verified the structure of the different
compounds. Twenty-two new phenolic compounds were char-
acterized from apple or apple pomace, especially naringenin,
luteolin and their derivatives. Factors such as pressure, temper-
ature, ethanol concentration and extraction time were found to
influence the subcritical extraction of polyphenols from apple
pomace (Adil et al., 2007). The phenolic profile of apple seeds
seems to be limited (Table 4) in comparison to apple skin (Lu
and Foo, 1998; Schieber et al., 2003). The effect of enzymatic
treatment on the yield of phenolic compounds was also investi-
gated under different extraction conditions (Zheng et al., 2008).
The pectinase treatment of apple pomace improved the extrac-
tion of polyphenolics compounds under optimized conditions of
enzyme reaction, i.e. pH 3.6, enzyme to pomace ratio of 12%,
37◦ C and an 11 h reaction period. This resulted in higher yields of
total phenolics (about 28%) and flavonoid content (about 50%)
in comparison to the control.
Polyphenols from apple extracts have been shown to inhibit
tumour-cell proliferation in vitro (Eberhardt et al., 2000). The
antioxidant properties of apple pomace polyphenols were eval-
uated using a β-carotene/linoleic acid system, free radical scav-
enging activity using DPPH (1,1-diphenyl-2-picryl-hydrazyl)
and superoxide anion radical scavenging activities by a cellular
xanthine/xanthine oxidase system as a superoxide source (Lu
and Foo, 2000). Epicatechin and quercetin 3-glycosides from
pomace showed the highest activity, while phloridzin exhibited
moderate activities in comparison to Vitamins C and E. Ex-
cept for phloridzin (0.60 EC50 ), all of the apple polyphenols
exhibited good DPPH-scavenging properties, two to three times
higher than vitamins C (0.35 EC50 ) or E (0.30 EC50 ). EC50 is the
amount of antioxidant necessary to decrease the initial DPPH
concentration by 50%. Apple pomace polyphenols were found
to be effective superoxide scavengers, in comparison to vita-
mins C and E; however, among themselves, procyanidins were
observed to be superior to quercetin 3-glycosides, chlorogenic
acid, 3-hydroxyphloridzin and phloridzin.
These studies revealed that the polyphenols responsible for
the antioxidant activity in apple are still present in the po-
mace, and can easily be extracted for food fortification or nu-
traceutical product development. Apple pomace can therefore
 PROCESSING OF APPLE POMACE FOR BIOACTIVE MOLECULES
become an inexpensive and readily available source of dietary
antioxidants.
Source of Biocatalyst
The intrinsic enzymes system of apple pomace had been em-
ployed for biocatalysis. Biocatalysis can be defined as the uti-
lization of natural catalysts, such as enzymes (either isolated
or intrinsic to the living cells) to perform chemical transforma-
tions on organic compounds. Since the 1980s there have been
a few research groups working on the production of volatile
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by North Carolina State University on 03/11/13
aldehydes and alcohols by apple peripheral tissues from in situ
or added linoleic acid or linolenic acid (Paillard, 1979; Ambid
and Fallot, 1980; Drawert et al., 1986). Almosnino and Belin
(1991) explored an apple pomace intrinsic enzymes system for
the bioconversion of polyunsaturated fatty acids in the produc-
tion of aromatic compounds. Fresh, oxidized (exposed to air)
and frozen Golden Delicious apple pomace (10 g each) was
used as an enzyme source for the chemical transformation of
technical linoleic acid (75% linoleic acid) in 150 mL of McIl-
vaine buffer reaction volume using Tween 80 as an emulsifier
(37.5 µl). The reaction was carried out at 18◦ C for 4 h. Sulphur
dioxide and ascorbic acid were used to preserve and reduce
the browning of the apple pomace (caused from oxidation by
polyphenol oxidase). Micronized fresh apple pomace preserved
For personal use only.
with 60 p.p.m. of SO2 , and 500 p.p.m. of ascorbic acid was found
to increase the yield of hexanal and 2,4-decadienal by 56% and
95%, respectively. The endogenous enzymes system contained a
lipoxygenase that converted linoleic acid to 9,13-hydroperoxide
(Kim and Grosch, 1979) and these intermediates were further
catalyzed by hydroperoxide lyase, resulting in the formation of
volatile compounds, i.e. hexanal and 2,4-decadienal (Schreier
and Lorenz, 1982; Hatanaka et al., 1986). Hexanal formation
required both the lipoxygenase and hydroperoxide lyase en-
zymes, while formation of 2,4-decadienal only depended on
the presence of lipoxygenase for conversion into hydroperox-
ides (Almosnino and Belin, 1991). Later, the process was scaled
up in 5 L and 10 L Trimix reactors, in a 2 L buffered medium
containing 33% or 50% apple pomace and 0.5% or 2% linoleic
acid (Almosnino et al., 1996). The biocatalysis, occurring dur-
ing biochemical transformation, was monitored by the quantifi-
cation of hexenal (reflecting the expression of the double en-
zyme system, i.e. lipoxygenase and hydroperoxide lyase) and
2,4-decadienal production (reflecting the expression of lipoxy-
genase and β-scission decomposition). The authors found that
the reaction at 25◦ C, with continuous oxygen flow to vessel
headspace, resulted in high productivities of both aromatic com-
pounds. The best hexenal yield (160 mg kg−1 ) was obtained in
an alkaline pH (9.0) after 48 h, exhibiting the pH optima of
the enzyme complex. However, both the enzymes appeared to
be unaffected by acidic conditions in reaction buffers pH 1.4–
4.8.
Besides enzyme systems, apple pomace also contains min-
eral elements such as iron, zinc, copper, calcium and magnesium
(Table 2). As these micro-elements act as cofactors in many
293
biosynthetic processes, they can be investigated for new appli-
cations either in biotransformations or bioconversions.
Pigment Molecules
Oxidation of polyphenols by polyphenoloxidase leads to the
development of yellow–orange coloured juices or ciders during
processing in the presence of oxygen, which has a direct correla-
tion with the concentrations of hydroxycinnamic acid, flavanols
and dihydrochalcones, i.e. phloridzin and phloretin xylogluco-
side (Lea, 1984; Nicolas et al., 1994). Phloridzin is a phenolic
compound present in apple pomace and its enzymatic oxida-
tion has been shown to produce yellow products (Lu and Foo,
1997; Ridgway et al., 1997; Schieber et al., 2003). Phloridzin,
using apple leaf enzyme systems, was first converted into 3-
hydroxyphloridzin and later into the corresponding o-quinones
(Raa and Overeem, 1968). A brown reddish colour was ob-
tained, and it was proposed that these products resulted from
the phloroglucinol nucleus and quinones in head-to-tail oxida-
tive coupling reactions. Recent development in instrumentation
and analytical techniques has led to the identification and gener-
ation of structural information on compounds such as phloridzin
oxidation products (POP). With the help of mass spectrometry
and NMR, monomeric structures resulting from oxidation and
intramolecular oxidative coupling with simultaneous ring rearo-
matisations of POP to a colourless compound (POPi), which
were then converted to yellow pigment products (POPj), were
elucidated (Le Guernévé et al., 2004). The oxidation of phlo-
ridzin by mushroom polyphenol oxidase in POP production of
yellow pigment, as an alternative to tartrazine for application as
a food dye, was evaluated (Guyot et al., 2007). Mushroom PPO
was added to pre-incubated phloridzin (dissolved in phosphate
buffer 100 mM, pH 6.5) at 30◦ C. The reaction time was 0–52 h,
and the solution was immediately stabilized by the addition of
200 µL of o-phosphoric buffer and 100 µL of sodium fluoride
solution. They studied oxidative product formation kinetics, the
effect of enzyme concentration, the colouring power of POPj,
stability as influenced by pH and temperature, and radical scav-
enging activities. The POPj molecule exhibited strong colouring
power, and 30 mg L−1 was found enough to reach half saturation
of the colour at pH 3. Researchers observed that an acidic pH
(2.2–5.0) had no pronounced effect on the colour characteris-
tics; however, an alkaline pH (≥ 6.0) resulted in the formation
of orange pigment from the yellow. the free radical scavenging
activity of phloridzin and its oxidative products was also exam-
ined. The studies revealed that phloridzin and POPj had a very
weak antioxidant activity, while POPi had similar activity to that
of epicatechin, ascorbic acid and Trolox. A similar free radical
scavenging activity of phloridzin was observed by Lu and Foo
(2000). The food and cosmetic industries are always looking for
new highly water-soluble yellow pigments and the phloridzin
pigment could be an alternative food dye. However, significant
research efforts would still be required to make this a viable
process on an industrial scale.
 294 S. BHUSHAN ET AL.
CONCLUSIONS
The processing of apple pomace and other industrial food
residues for extraction of bioactive molecules can play an im-
portant role in efficient bioresource utilization and can provide a
method for the nutritional enrichment of foods at a low cost. As
per the reviewed literature, target molecules from apple pomace
are dietary fibre, pectin (especially tailored polysaccharides with
unique functional properties) and, more importantly, natural an-
tioxidants. Dietary fibres from fruit residues have more benefi-
cial effects on human health than cereals owing to the presence
Critical Reviews in Biotechnology Downloaded from informahealthcare.com by North Carolina State University on 03/11/13
of associated bioactive molecules. Polysaccharides tailored for
unique functional properties can be employed for the develop-
ment of industrial molecules such as bioabsorbents, biopolymers
or compounds for effluent treatment.
A lot of interest has been generated regarding natural antiox-
idants, owing to their role as a free radical scavenger, as these
are associated with a number of degenerative diseases such as
cancer and atherosclerosis. The recovered antioxidants from ap-
ple pomace showed good free radical scavenging activities, and
hence could be used as a nutraceutical and a food supplement.
In addition to these bioactive molecules, the biocatalytic prop-
erties of apple pomace can be explored for the conversion of
low value substrates to high value product formation. However,
further R&D efforts are required for product development and
For personal use only.
for the assessment of their safety and stability with respect to
associated human health benefits. It is also worth mentioning
that apple crops are heavily sprayed with chemicals owing to
their susceptibility to insect and pathogen attacks, and therefore
a toxicity analysis of apple pomace will be required prior to
processing.
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