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Effect of pasteurization and storage on

bioactive components of Aloe vera gel

Article in Nutrition & Food Science · March 2013

DOI: 10.1108/00346651311313553

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Bioactive
Effect of pasteurization and components of
storage on bioactive components Aloe vera gel
of Aloe vera gel
175
Hamed Saberian, Zohreh Hamidi-Esfahani and Soleiman Abbasi
Department of Food Science and Technology, Tarbiat Modares University,
Tehran, Iran

Abstract
Purpose – Aloe vera gel has nutritional and therapeutic properties due to presence of bioactive
components. The aim of this study is to investigate the effect of pasteurization and storage time at
different temperatures on some bioactive components of Aloe vera gel juice.
Design/methodology/approach – Aloe vera gel juice (Aloe barbadensis Miller) was pasteurized at
908C for 1 min and stored up to 30 days at 4 and 258C. The effect of pasteurization and storage time on
glucomannan, vitamin C, DPPH inhibition (percent) and color of the juice was evaluated.
Findings – The results showed that pasteurization reduced vitamin C content and antioxidant
activity 16% and 57%, respectively. During storage at 4 and 258C, vitamin C and glucomannan
contents reduced from 84.47 to 54.96 and 46.82 mg vitamin C/100 g dm and from 2.11 to 1.77 and
1.71 g/L, respectively. DPPH inhibition (percent) and Browning index (BI) increased significantly at
both storage temperatures, which was more intensive at 25 than 48C.
Originality/value – This paper is believed to be the only one which investigates the effect of thermal
pasteurization and storage time at different temperatures on some bioactive components of
Aloe vera gel juice.
Keywords Aloe vera, Pasteurization, Storage, Functional properties, Temperature, Nutrition
Paper type Research paper

1. Introduction
More than 400 species of Aloe plants have been identified, but a few of them have
medicinal or economic value (Shelton, 1991). Aloe barbadensis Miller, also known as
Aloe vera (L.) Burm. f., or simply Aloe vera, is the most widely used and commercially
available Aloe because of its nutritional and therapeutic properties (Reynolds, 2004)
and has often been referred to as the miracle plant, healing plant, wand of heaven and
plant of life (Olariu, 2009). It belongs to the lily (Olariu, 2009) and originated from warm
and dry climates of Africa, and then transplanted to the Far East and Western
hemispheres (Agarry et al., 2005).
The Aloe vera leaf has two major liquids including a yellow exudate produced by
the bundle sheath cells of skin containing mainly anthraquinones, which are used as
cathartics, and parenchyma cells containing mucilaginous gel (Femenia et al., 2003).
This clear inner gel has over 75 bioactive components especially phenol compounds,
vitamins, bioactive polysaccharide (Reynolds, 2004). Many of the medicinal effects of
gel have been attributed to its bioactive polysaccharide, namely glucomannan
(Hamman, 2008), but most researchers claim that the observed effects may be due to Nutrition & Food Science
Vol. 43 No. 2, 2013
pp. 175-183
q Emerald Group Publishing Limited
The authors are grateful to the Department of Food Science and Technology in Tarbiat Modares 0034-6659
University (Tehran, Iran) for financial support of the project. DOI 10.1108/00346651311313553
NFS the synergistic actions of the 75 known ingredients (Vega-Galvez et al., 2011). Aloe vera
43,2 gel has been utilized as a resource of functional foods, especially in healthy drinks and
beverages, a medicine and also in the cosmetic and toiletry industries (Ramachandra
and Srinivasa, 2008).
Conventional pasteurization is the most common processing technique for preserving
food products such as Aloe vera gel (Qian et al., 2005). In addition to the intended effects
176 on pathogenic and spoilage microorganisms and deteriorative enzymes, it is known that
thermal processing may affect food quality (Farnworth et al., 2001). Aloe vera gel
contains unstable compounds such as glucomannan, and if it is not stabilized against
Aloe bacterial contamination and endogenous hydrolyzing enzymes, this bioactive
component undergoes rapid degradation (Eberendu and McAnalley, 1996).
The aim of this study was to investigate the effect of pasteurization and storage
time at different temperatures on some bioactive components of Aloe vera gel juice,
especially its glucomannan.

2. Materials and methods

2.1 Materials
All reagents were purchased from Merck (Merck KGaA, Darmstadt, Germany) and all
measurements were done in triplicate.

2.2 Sample preparation

Leaves of Aloe vera (Aloe barbadensis Miller) were provided by experimental farm of
Tarbiat Modares University, Tehran, Iran. The homogenous leaves were selected
according to size, ripeness, color, and freshness. Aloe latex (the yellow-colored liquid)
was extracted by cutting the base of the leaves and allowing them to drain for 1 hour.
The epidermis was separated; pulp cut into small pieces and blended in a mixer. The
blended sample was filtered through a four-fold cloth to remove every fiber and obtain
a clear gel. pH of gel was adjusted to 3.5 with citric acid and kept under cool conditions
in a refrigerator (48C) until thermal processing.

2.3 Thermal treatment and storage

Aloe vera gel juice (about 20 mL) was transferred into Pyrex tubes and thermally
treated (pasteurized) in a water bath at 908C for 1 min to ensure the inactivation of
spoilage microorganisms (Qian et al., 2005). Samples were immediately cooled in a
water bath with ice. Pasteurized samples were stored at 4 and 258C for 30 days.
Microbiological and quality analyses were performed after processing at intervals
during the storage period.

2.4 Microbiological analysis

Samples were analyzed for numbers of mesophilic aerobic microorganisms, acid
resistant bacteria, lactic acid bacteria, moulds and yeasts.

2.5 Quality parameters

2.5.1 Physicochemical determinations. pH was directly measured using a pH-meter.
Moisture content was determined by AOAC (1990) method no. 934.06.
2.5.2 Determination of glucomannan. The Aloe vera polysaccharide was
determined according to Eberendu and McAnalley (1996) with some modifications.
First, Aloe vera gel samples were filtered through a Whatman 0.45 mm syringe filter. Bioactive
1 mL of every sample was diluted four times and transferred into test tubes. Next, 5 mL of components of
0.2 M NaOH and then 1 mL of 2 £ 102 4 M aqueous Congo Red solution were added and
slightly mixed. Finally, the absorbance of samples was measured at 540 nm using Aloe vera gel
spectrophotometer.
2.5.3 Vitamin C (L-ascorbic acid). L-ascorbic acid was determined by the 2,
6 dichlorophenol-indophenol titration method according to AOAC (2000). A total of 177
10 þ 0.1 g of Aloe vera gel was weighed and diluted to the volume of 50 mL. Vitamin C
content is expressed as mg vitamin C/100 g d.m.
2.5.4 Determination of DPPH radical scavenging activity. The free radical scavenging
activity of samples was determined using 2,2,-diphenyl-2-picryl-hydrazyl (DPPH)
method according to Tezcan et al. (2009) with some modifications. An aliquot of 1 mL of
0.006 percent DPPH radical in ethanol was added to a test tube with 1 mL of the sample.
The reaction mixture was mixed on a vortex mixer for 30 s and left to stand at room
temperature in the dark for 30 min. The absorbance was measured at 517 nm using the
spectrophotometer. Ethanol was used to calibrate the spectrophotometer. The control
sample was prepared without adding the sample. Total antioxidant activity was expressed
as the percentage inhibition of the DPPH radical and determined by equation (2):
DPPH inhibition ð%Þ ¼ ð1 2 ðAbssample =Abscontrol ÞÞ ð1Þ
where Abs is absorbance.
2.5.5 Color. The sample color was measured before pasteurization and during
storage by Hunter – Lab ColorFlex, A60-1005-654 45/0 model colorimeter. Color value
was expressed as L * (whiteness or brightness/darkness), a * (redness/greenness) and
b * (yellowness/blueness). Also, browning index (BI) (equation (2)) was calculated from
the Hunter L *, a *, b * values and used to describe the color changes during storage
(Zakipour and Hamidi-Esfahani, 2011):
BI ¼ ½100ðX 2 0:31Þ
=0:17 ð2Þ
X¼ ða* þ 1:75 L* Þ=ð5:645 L* þ a* 2 3:012 b* Þ ð3Þ

2.6 Statistical analysis

Results were analyzed by analysis of variance using SAS 9.1 Statistical Software and
the LSD test with 95 percent confidence interval was used to compare the means of the
tests.

3. Results and discussion

3.1 Physicochemical properties
The initial moisture content of Aloe vera gel juice was about 99.27 percent. Since the
moisture content of this product is very high, so other components are in very low
concentrations.

3.2 Microbiological assays

Microbiological assays showed that all pasteurized samples during storage at two
temperatures exhibited appropriate levels of microbiological safety according to World
Health Organization (WHO, 1999). The results are in agreement with the previous
reported results in literature for acidic fruit juices (Qian et al., 2005).
NFS 3.3 Quality assessment
43,2 3.3.1 Glucomannan. An assay for qualitative and quantitative determination of
glucomannan, a bioactive polysaccharide which is the special polysaccharide of
Aloe vera gel (Eberendu and McAnalley, 1996) was used. This method uses a complex
agent, namely Congo Red to react with b 2 1, 4 link of glucomannan present in the Aloe
vera gel and gives a color change. The color change is compared with standard color
178 changes and the difference between color changes is dependent on the amount of
glucomannan.
Glucomannan could not be spectrophotometrically measured before thermal
processing because after NaOH addition, it precipitated alone or along with other
colloids, which reached their isoelectric point and formed the yellow sediment. However,
after pasteurization, this phenomenon did not occur because the thermal treatment
probably changed the electrostatic charge of colloids, which prevented the sediment
formation after NaOH addition. The results showed that the amount of glucomannan
after pasteurization (time ¼ first day) was 2.11 þ 0.035 g/L (288 þ 4.8 mg
glucomannan/g d.m. of fresh gel juice) that is similar to the glucomannan content
(268 mg glucomannan/g d.m. of fresh Aloe filets) reported by Femenia et al. (2003), but
higher than the glucomannan content (113-139 mg glucomannan/g d.m. of fresh Aloe
filets) reported by Rodriguez-Gonzalez et al. (2011). It seems logical that Aloe vera gel
without fibers has a higher amount of glucomannan than Aloe vera fillet because the
fillet has a lot of fibers, but in the clear gel fibers were removed. The bioactive
polysaccharide of Aloe vera gel decreased significantly during storage at both
temperatures, which this decline was much higher at 258C than 48C (Figure 1).
After 30 days storage at 4 and 258C, the glucomannan content decreased by
8.30 and 26 percent, respectively, indicating three times higher losses at 258C than at 48C.
Xiu et al. (2006) reported that enzymes in Aloe vera gel juice lead to the degradation
of the polysaccharide. After pasteurization of Aloe vera gel, its enzymes as well as
some bacteria and fungi probably remained and produced enzymes that could

Figure 1.
Effect of storage time
at 4 and 258C on the
glucomannan
hydrolyse glucomannan. Meanwhile, microbes can grow and produce enzymes at 258C Bioactive
better than 48C resulting higher losses of the glucomannan. components of
3.3.2 Vitamin C. Figure 2 shows the effect of thermal treatment and storage times on
vitamin C content. Before pasteurization, the initial vitamin C content (time ¼ 0) of Aloe vera gel
Aloe vera gel was 84.47 þ 1.76 mg/100 g d.m which is lower than the amount
(126.37 mg/100 g d.m) reported by Vega-Galvez et al. (2011). The results indicated that
there was a significant difference ( p , 0.05) in vitamin C content between thermally 179
treated (time ¼ first day) and untreated (time ¼ 0) samples so that vitamin C retention
after pasteurization was 84 percent. Ascorbic acid is a thermo labile nutrient, but
resistant to low pH. Since vitamin C is generally degraded by oxidative processes which
are stimulated in the presence of light, oxygen, heat, peroxides and enzymes, vitamin C
content decrease during pasteurization could be due to the high temperatures applied
during pasteurization (Burdurlu et al., 2006). Vitamin C retention has been used as an
indicator of fruit juice quality as well as a shelf-life marker for chilled fruit juices. When
vitamin C retention decreases to 50 percent of its initial amount, shelf-life ends
(Morales-de la Pena et al., 2011). The analysis of vitamin C content during storage
showed that there is a soft deterioration at both temperatures. Vega-Galvez et al. (2011)
reported that vitamin C of Aloe vera gel decreased during storage at refrigerator.
The retention of vitamin C after 30 days of storage at 48C and 258C were 65 and
55 percent, respectively, indicating that vitamin C degradation at room temperature is
higher than refrigerated temperature. Ascorbic acid retention in citrus juices after
storage at 288C, 378C and 458C was about 54.5-83.7, 23.6-27 and 15.1-20.0 percent,
respectively, due to the formation of hydroxyl methyl furfural (HMF) which is one of
the decomposition compounds of ascorbic acid degradation (Burdurlu et al., 2006).
Similar results were reported by Esteve et al. (2005) that ascorbic acid of a Spanish
orange juice varied with storage time more significantly at 108C than 48C.
3.3.3 DPPH inhibition. Antioxidant capacity of fruit juices is related to the
composition and concentration of bioactive compounds such as vitamins, phenols,
carotenoids or flavonoids (Morales-de la Pena et al., 2011). The influence of thermal
treatment on DPPH inhibition is shown in Figure 3. Initial percentage of DPPH

Figure 2.
Effect of storage time
at 4 and 258C on
vitamin C content
NFS
43,2

180

Figure 3.
Effect of storage time
at 4 and 258C on DPPH
inhibition (percent)

inhibition (time ¼ 0) was 54 þ 0.85 percent and after pasteurization (time ¼ first day),
it decreased to 23 þ 2.8 percent, less than 50 percent of initial inhibition percentage.
Pasteurization of grapefruit provoked a significant decrease ( p , 0.05) of DPPH
inhibition and its loss was 40 percent (Igual et al., 2010).
The main cause of depletion of natural antioxidants is generally thermal treatments
(Anese et al., 1999). As shown in Figure 3, during storage at 4 and 258C, the percentage
of DPPH inhibition increased significantly ( p , 0.05), which this increase was much
higher at 258C than 48C. After 30 days, DPPH inhibition (percent) at 4 and 258C were
28.9 þ 1.34 percent and 36.7 þ 0.45 percent, respectively, (Figure 3). The increase in
the activity may be explained by the strong tendency of polyphenols to undergo
polymerization reactions, whereby the resulting oligomers possess larger areas
available for charge delocalization (Morales-de la Pena et al., 2011). Also Klimczak et al.
(2007) reported that increase in the antioxidant activity is usually ascribed to
Maillard’s reaction products.
3.3.4 Color. Color is the first quality factor of a food that a consumer appreciates and
has an important influence on its acceptance. Color is also an indicator of changes of
foods that occur during storage or processing (Esteve et al., 2005). As shown in Figure 4,
BI of Aloe vera gel juice stored at both temperatures increased but this increase was
considerably higher at 25 than 48C. In some fruit juice concentrates, the accumulation of
brown color during thermal processing is attributed to enzymatic browning, but during
storage, it is mainly due to nonenzymatic browning, which include caramelization,
ascorbic acid degradation and Maillard reaction (Babsky et al., 1986). HMF is one of the
decomposition products of ascorbic acid, which are suggested to act as a precursor of
brown pigments. Burdurlu et al. (2006) reported that after an eight-week storage, HMF
content of citrus juice concentrates at 458C increased more significantly than 288C which
had a significant correlation with the ascorbic acid loss. The addition of Aloe vera gel
into film-forming chitosan solution at higher levels yielded blend films with darker
appearance (brown), which can be attributed to the anthraquinones oxidation from the
 Bioactive
components of
Aloe vera gel

181

Figure 4.
Effect of storage time
at 4 and 258C on BI

skin, because they are difficult to be completely removed (Khoshgozaran-Abras et al.,

2012). Therefore, it seems that the most important reasons for more color changes at 258C
than 48C were more vitamin C degradation, Mailard reaction and anthraquinones
oxidation at 258C.

4. Conclusions
Pasteurization performed at 908C for 1 min could safe Aloe vera gel juice and this
safety remained stable during storage. During storage at 4 and 258C glucomannan
significantly decreased due to the hydrolysis of bioactive polysaccharide by enzymes.
Vitamin C content decreased because of ascorbic acid degradation and Mailard
reaction. DPPH inhibition percentage decreased by pasteurization due to the effect of
high temperature, but increased during storage, which can be ascribed to Maillard’s
reaction products. Also BI was increased at both storage temperatures due to vitamin C
degradation, Mailard reaction and anthraquinones oxidation. The rate of changes was
much more considerable at 25 than 48C due to direct influence of increasing
temperature on the reactions’ speed. Regarding the bioactive polysaccharide of
Aloe vera gel, which is the main component of the gel and its significant decrease
during storage at 25 than 48C as well as color changes that were more remarkable at
25 than 48C, it can be concluded that the characteristics of Aloe vera gel were better
retained during storage at 48C rather than 258C.

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Corresponding author
Zohreh Hamidi-Esfahani can be contacted at: hamidy_z@modares.ac.ir

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