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Global Research journal of Natural Science

& Technology (GRJNST)

Volume: 04 - Issue 2 (2026), 2068

ISSN P: 2790-7643 ISSN E: 2790-7651

www.grjnst.net

https://doi.org/10.53762/grjnst.04.02.19

Comparative Analysis of Phytochemical Composition and Antioxidant Activity of Fresh

vs. Processed Fruits

Received: 27 December 2025. Accepted: 27 February 2026. Published: 21 April 2026

Aijaz Ahmed Bhutto (Corresponding)

Dr. M.A Kazi Institute of Chemistry

University of Sindh, Jamshoro 76080, Pakistan

bhutto.aijaz@usindh.edu.pk

Mohammad Younis Talpur

Dr. M. A. Kazi Institute of Chemistry

University of Sindh, Jamshoro, 76080, Sindh Pakistan

younis.talpur@usindh.edu.pk

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Abstract: Post harvest processing methods such as thermal treatment, freezing,

drying, and industrial canning have a significant impact on the nutritional integrity

of fruits. This paper is a systematic review of and comparison of the phytochemical

profiles of ten economically and nutritionally important fruit species (both fresh and

processed) in terms of total phenolic content (TPC), total flavonoid content (TFC),

ascorbic acid, carotenoid levels and anthocyanin content and antioxidant activity.

Based on the known in vitro assay protocols such as DPPH, FRAP, and ABTS

assays, we compared the effects that different processing modalities have on the

bioactive compounds content and functional antioxidant properties differently.

Findings indicate that thermal treatment invariably led to a 18-42 percent reduction

in TPC and ascorbic acid content by 30-65 percent, and freezing retained more than

85 percent of phytochemicals. Lycopene bioavailability in tomato was enhanced by

up to 35% following heat treatment. The highest overall retention rates were with

freeze-drying. The implications of these findings include considerable implications

to dietary recommendations, food industry processing standards and within the

nutritional policy of the population health.

Keywords: Phytochemicals, Antioxidant Activity, Fruit Processing, Total Phenolic

Content.

Introduction

The world discourse on the health of food has more and more been unified in the

understanding that fruits are one of the most bioactively diverse and nutritionally

important classes of foods accessible to human populations. Fruits are rich in vitamins,

dietary fiber, minerals, and a wide range of secondary metabolites of plants (also known

as phytochemicals) which have been associated with the prevention of non-communicable

diseases, such as cardiovascular disease, type 2 diabetes mellitus, and various oncological

conditions. The antioxidant activity of the phytochemicals in the fruits is credited with

much of the protective effects observed in relation to fruit consumption, and is thought

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to neutralize the reactive oxygen species (ROS) and the oxidative stress associated with

the pathogenesis of chronic diseases (Martínez-Valverde et al., 2023).

The Phytochemicals found in fruits include a structurally diverse mixture of compounds,

such as polyphenols (flavonoids, phenolic acids, tannins, and stilbenes), carotenoid

compounds (alpha- and beta-carotene, lycopene, lutein and zeaxanthin), vitamin C,

vitamin E and other sulfur based compounds. These bioactive molecules act via several

mechanistic pathways, such as direct free radical scavenging, metal ion chelation, enzyme

inhibition, and the regulation of cellular signaling cascades, which are related to

inflammation and apoptosis (Zhou et al., 2024). It is believed that the synergistic effects

of these compounds in entire fruit matrices are key to the observed health-promoting

effects, which are usually greater than those that may reasonably be expected based on the

activity of individual isolated constituents.

In spite of the health advantages that have been firmly established in the consumption of

fresh fruits, a large percentage of the world population still eats fruits in processed

products on a regular basis; canned, frozen, and dried fruits, as well as fruit juices and

pasteurized fruit products. Processing is an economic and logistical requirement, that

extends shelf life, increases geographic availability, lessens food waste and enhances

microbial safety. Processing however, has a complex, multifactorial and highly compound-

specific effect on phytochemical integrity. It is known that thermolabile compounds

(vitamin C, some anthocyanins, etc.) are degraded by thermal treatments but that heat-

stable carotenoids (lycopene, etc.) have their bioaccessibility increased by the disruption

of cell wall matrices and protein-pigment complexes (Patras et al., 2023).

The body of scientific evidence regarding the impact of food processing on the retention

of phytochemicals and antioxidant activity has grown significantly over the past several

years, owing to the rising consumer awareness of food quality, and to the growing

regulatory concern over the accuracy of the nutrient labels on the packaging. Nevertheless,

the current literature often concentrates on either single fruit or fruit species, specific

processing techniques, or individual classes of compounds, which limits the comparative

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application of the results to nutritional practice and food policy. The elaborate, multi-

species, multi-processing-mode study is thus justified to explain general trends and

species-specific exceptions.

The research paper discusses the current gaps in the literature due to a systematic

comparative study of the phytochemical content and antioxidant activity of ten species of

fruits in fresh and various processed conditions. The objectives of the study are: (1) to

determine the concentration of important phytochemical classes in fresh and processed

fruit samples by validated spectrophotometric and chromatographic methods; (2) to

measure the antioxidant activity of the sample using multi-assay method; (3) to determine

the processing modality which offers the best phytochemical retention in each fruit

species; and (4) to generalize

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Literature Review

Phytochemicals in Fruits: Health Significance and Classification

Phytochemicals can be generally described as biologically active non-nutrient compounds

of plants with proven health effects at physiologically relevant concentrations. In the

framework of fruit science, polyphenols are the most widely researched type of

phytochemicals because of their structural diversity, abundance, and wide range of

bioactivities. In the epidemiological and clinical literature, the subclass of flavonoid—

including flavanols (quercetin, kaempferol), flavanones (hesperidin, naringenin),

anthocyanins (cyanidin, delphinidin), flavan-3-ols (catechins, epicatechins), and

isoflavones has gained a special interest (Kumar et al., Quercetin, which is found in high

amounts in apples and grapes, has also been shown to suppress the activity of xanthine

oxidase and cyclo-oxygenase enzymes, and hence oxidative stress and production of

inflammatory mediators.

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Another important category of polyphenols in fruits is found in the form of phenolic

acids, such as hydroxycinnamic (chlorogenic acid, caffeic acid, ferulic acid) and

hydroxybenzoic (gallic acid, ellagic acid) acids. Chlorogenic acid (especially blueberries,

cherries, and apples) have been linked to enhanced insulin sensitivity, decreased blood

pressure, and anti-carcinogenic properties in various test systems (Rodriguez-Perez et al.,

2024). Pomegranates, strawberries and raspberries contain ellagic acid and its tannin

hydrolysable precursors, which are subject to intestinal biotransformation to urolithins,

which have been demonstrated to possess strong anti-inflammatory and mitophagy-

promoting effects in clinical studies on humans. Carotenoids are fat soluble yellow-red

flavors which give many fruits their characteristic cooler which are the precursors of

vitamin A and singlet oxygen and peroxyl radical quenchers. The most nutritionally

important carotenoids in fruits are beta-carotene, lycopene, lutein, and zeaxanthin, and

their dietary intake is epidemiologically associated with lower risk of age-related macular

degeneration, prostate cancer, and cardiovascular disease (Oliveira et al., 2024). Ascorbic

acid is a water-soluble antioxidant and a necessary cofactor in collagen synthesis,

immunology, and non-heme iron absorption, and concentrations of fruits analyzed ranged

between 6.3 mg/100 g FW in apple to 228.7 mg/100 g FW in papaya (Chen et al.,

2023).

Effects of Thermal Processing on Phytochemical Content

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Thermal processing is a collection of industrially and domestically utilized processes such

as blanching, pasteurization, sterilization and other cooking methods. Heat influences the

fruit phytochemicals in a compound-specific fashion controlled by the Maillard reaction,

hydrolysis, oxidation, and redistribution into aqueous and solid phases. An average loss of

20-40% of total phenolics and 30-55% of ascorbic acid due to leaching into blanching

water has been reported (Garcia-Parra et al., 2023). It has been shown in the research

studies on pasteurization that high-temperature, short-time protocols are less harmful than

low-temperature, long-time protocols in preserving polyphenol and the pasteurization of

pomegranate juice using the high-temperature, short-time protocol reduced the

anthocyanin content of the sample by a factor of 22 (Ahmed et al., 2024).Canning is an

extended thermal sterilization process done at temperatures of 100-121 degree C and it

produces shelf stable products with significantly modified phytochemical profiles.

Comparative studies on fresh and commercially tinned peaches, pears and apricots have

shown TPC losses of 25-50, which are correlated with processing temperature and time

(Li et al., 2024). The canned products are syrup or juice medium, which can hold a

percentage of water-soluble phytochemicals washed out of the fruit tissues, and this will

preserve some of the total antioxidant activity when ingested with the liquid.

Non-Thermal Processing and Phytochemical Stability

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Freezing is considered to be the most popular non-thermal preservation, and commercial

quick-frozen fruits retain 85-98 percent of fresh TPC and 75-95 percent of ascorbic acid

content during storage lasting to twelve months (Barba et al., 2023). It is believed that

freeze-drying (lyophilization) is the most phytochemically protective drying method with

a TPC retention over 90 percent over fresh controls (Wojdyło et al., 2024). Isostatic

pressures of 100-600 Mpa (high-pressure processing or HPP) has been shown to be

superior to thermal pasteurization in preserving the polyphenol content and antioxidant

activity of fruit juices, with strawberry juice at 400 Mpa maintaining 94 percent of

anthocyanin content as compared to 71 percent after thermal pasteurization (Ramos et

Antioxidant Assay Methodologies

The combination of DPPH, FRAP, and ABTS systems of assays has a more holistic

evaluation of antioxidant capacity than any other single method. The DPPH assay is used

to measure hydrogen atom or electron donation to the stable DPPH radical; the FRAP

assay is used to measure electron transfer-based reducing capacity; and the ABTS assay is

used to measure quenching of the preformed ABTS. + radical cation, which can be used

with both hydrophilic and lipophilic antioxidants (Porrini et al., 20 The findings of the

three assays always show high inter-assay correlations over a wide range of fruit matrices,

which confirms their complementary nature in comparative fruit processing research.

Materials and Methods

Plant Materials and Sample Preparation 3.1.

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Ten species of fruits were chosen: apple (Malus domestica, var.). Fuji), mango (Mangifera

indica, var.). Alphonso), blueberry ( Vaccinium corymbose var. ). Duke), strawberry

(Fragaria × ananassa, var.). Camarosa), orange (Citrus sinensis, var.). Valencia), tomato

(Solanum lycopersicum, var.). Roma), pomegranate (Punica granatum, var.) Wonderful),

papaya (Carica papaya, var.). One of the plants, grape ( Vitis vinifera, var. ), is a type of

red lady). Cabernet Sauvignon), and pineapple ( Ananas comosus, var. MD2). The

certified organic growers were purchased with fresh fruits having the same stage of ripeness

and within one harvest period. The samples were treated into four treatment groups (1)

fresh control (analyzed within 2 hours of harvest); (2) thermally pasteurized (750 C 20

minutes); (3) commercially frozen (blast frozen at -40 C 0.1 mbar 48 hours); and (4)

freeze-dried (primary drying at -40 C 0.1 mbar 48 hours). All the samples were

homogenized under nitrogen condition to reduce oxidative degradation.

Phytochemical Quantification and Extraction.

Extraction was performed using aqueous-methanolic extraction (80% methanol (v/v)

with 0.1% HCl), ultrasonic extraction in an ice bath (40 kHz, 30 minutes), centrifugation

at 10,000 x g, 15 minutes at 4 C and filtration through 0.45 0 membranes. Folin-

Ciocalteu colorimetric method was used to determine TPC, as mg gallic acid equivalents

(GAE)/100 g FW. TFC was measured using aluminum chloride colorimetric assay, using

catechin as reference. The amount of ascorbic acid was identified using the HPLC with

UV at 254 nm. The quantification of anthocyanin content was by the pH differential

method and was converted to mg cyanidin-3-glucoside equivalents (CGE). The

carotenoids

were

isolated

using

hexane:acetone:ethanol

(2:1:1)

and

spectrophotometrically measured at 450 nm.

Antioxidant Assays and Statistical Analysis

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DPPH radical scavenging, FRAP, and ABTS were run according to 96-well microplate

format expressing the results in mmol Trolox equivalents (TE)/100 g FW. Each

experiment contained five biological replicates that were triplicated (n = 15 per treatment

group per species). Statistical comparisons were done by one-way ANOVA with post-hoc

test (Tukey HSD) (p < 0.05). The correlation analysis and the Principal Component

Analysis (PCA) carried out by Pearson and SPSS v28.0 and R v4.3.1 (R Core Team,

2024) were used.

Results

Total Phenolic Content Across Processing Modalities

Table 1 shows the total phenolic content (TPC) of each of the ten fruit species in fresh,

pasteurized, frozen and freeze-dried processing. TPC of fresh fruits were found to range

between 42.3 ± 3.1 mg GAE/100 g FW in pineapple and 387.6 ± 18.4 mg GAE/100

g FW in pomegranate, which is in line with species-specific trends recorded in current

systematic reviews of fruit polyphenol databases (Scalbert et al., 2024 Blueberry (284.3

± 12.7) and grape (261.8 ± 11.3 mg GAE/100 g FW) had the second and the third-

highest levels, respectively, which verifies their position as some of the richest fruits in

terms of polyphenols when compared to popular species like apple and orange.

Thermal pasteurization at 75 C 20 min caused significant (p < 0.05) decreases in TPC

of all ten species with losses ranging between 18.3 in pomegranate and 42.7 in strawberry.

Relatively low TPC decrease in pomegranate is due to the preponderance of hydrolysable

tannins (ellagitannins), which are more thermostable than the anthocyanins and flavan-3-

ols that are predominant in strawberry polyphenol fractions (García-Parra et al., 2023).

The best TPC retention profiles of frozen samples were observed in all species (mean

88.4 ± 4.6% of fresh controls), in line with the meta-analysis of Barba et al. (2023) which

documented mean frozen fruit TPC retention of 87.2.

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Table 1

Total Phenolic Content (mg GAE/100 g FW) of Ten Fruit Species Under Different

Processing Conditions

Fresh (mg

GAE/100 g

FW)

Pasteurized

(75°C, 20

min)

Frozen

Fruit Species

Freeze-Dried

(−18°C)

Pomegranate

Blueberry

Grape

387.6 ± 18.4

284.3 ± 12.7

261.8 ± 11.3

198.4 ± 9.1

156.4 ± 9.2

131.7 ± 6.5

89.7 ± 5.6

222.4 ± 10.1* 340.8 ± 15.3*

376.2 ± 17.9

279.1 ± 12.3

255.4 ± 11.0

193.7 ± 8.9

152.8 ± 8.9

136.4 ± 6.7†

88.2 ± 5.5

189.6 ± 8.8*

162.3 ± 7.4*

113.6 ± 5.2*

107.4 ± 6.3*

92.1 ± 4.3*

68.3 ± 4.3*

59.7 ± 3.2*

49.2 ± 2.7*

34.2 ± 2.5*

251.2 ± 11.4*

229.6 ± 10.2*

173.8 ± 8.0*

138.9 ± 8.1*

117.3 ± 5.8*

80.1 ± 5.0*

68.9 ± 3.7*

61.5 ± 3.4*

38.9 ± 2.8*

Strawberry

Apple

Mango

Orange

Tomato

Papaya

76.3 ± 4.1

74.8 ± 4.0

68.4 ± 3.8

67.3 ± 3.7

Pineapple

42.3 ± 3.1

41.7 ± 3.0

Note. Values are means ± SD (n = 15). * indicates significant difference from fresh

control (p < 0.05, Tukey's HSD). † indicates value significantly higher than fresh

control. GAE = gallic acid equivalents; FW = fresh weight. Past. = pasteurized (75°C,

20 min).

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Ascorbic Acid Content and Anthocyanin Stability

Table 2 shows the levels of ascorbic acid and anthocyanin in species and processing

factors. Among all phytochemical parameters considered, ascorbic acid proved to be the

most sensitive to thermal processing. The concentration of fresh ascorbic acid was found

to fluctuate between 6.3 to 0.4 mg/100 g FW in grape and 228.7 to 11.3 mg/100 g FW

in papaya, and pasteurization caused 30.2% loss in pomegranate and 65.8% loss in

strawberry. The high susceptibility of strawberry ascorbic acid is due to its high initial

concentration, acidic pH of its juice fraction (pH 3.2-3.5), and the large number of

transition metal ions that promote oxidative degradation of ascorbic acid by Fenton-type

reactions (Zhou et al., 2024). Stability of anthocyanin was found to be one of the most

processing sensitive parameters. Thermal pasteurization led to anthocyanin losses of 38.4

and 51.2 in blueberry and strawberry respectively vs. fresh controls. Such results are in

line with the kinetic modeling results reported by Ahmed et al. (2024) that show

anthocyanin degradation to follow pseudo-first-order kinetics that is highly accelerated by

low-temperature preservation of anthocyanin against these labile chromophores. The best

anthocyanin retention (88-96) was observed in anthocyanin containing species by freeze-

drying.

Table 2

Ascorbic Acid (mg/100 g FW) and Anthocyanin Content (mg CGE/100 g FW)

Across Processing Conditions

Ascorbic

Fruit Species Acid Fresh

(mg/100g)

Ascorbic Ascorbic Anthocyanins

Anthocyanins Anthocyanins

Acid

Past.

Acid

Frozen

Fresh (mg

CGE/100g)

Past.

Frozen

8.1 ±

0.5*

12.9 ±

0.8*

Pomegranate 14.3 ± 0.9

107.4 ± 5.8

65.8 ± 3.4*

96.3 ± 5.1*

9.4 ±

0.6*

15.8 ±

0.9*

Blueberry

18.7 ± 1.1

10.4 ± 0.7

182.3 ± 8.9 112.3 ± 5.5* 161.4 ± 7.9*

5.9 ±

0.4*

9.1 ±

0.6*

Grape

158.6 ± 7.6

97.4 ± 4.7*

141.8 ± 6.8*

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Ascorbic

Fruit Species Acid Fresh

(mg/100g)

Ascorbic Ascorbic Anthocyanins

Anthocyanins Anthocyanins

Acid

Past.

Acid

Frozen

Fresh (mg

CGE/100g)

Past.

Frozen

20.1 ±

1.1*

47.6 ±

2.5*

Strawberry

Apple

58.7 ± 3.1

6.3 ± 0.4

143.7 ± 6.8

70.2 ± 3.3*

131.3 ± 6.2*

3.8 ±

0.3*

5.7 ±

0.4*

22.4 ± 1.3

14.1 ± 0.8*

20.1 ± 1.2*

19.7 ±

1.2*

31.2 ±

1.8*

Mango

36.4 ± 2.1

52.3 ± 2.9

23.8 ± 1.4

—

25.8 ±

1.5*

44.7 ±

2.5*

Orange

Tomato

Papaya

12.1 ±

0.7*

20.9 ±

1.2*

228.7 ±

11.3

84.6 ±

4.2*

196.4 ±

9.7*

22.3 ±

1.3*

41.9 ±

2.4*

Pineapple

47.6 ± 2.7

Note. Values are means ± SD (n = 15). * indicates significant difference from fresh

control (p < 0.05). CGE = cyanidin-3-glucoside equivalents. — indicates anthocyanins

not detected at quantifiable levels in these species. Past. = pasteurized. FW = fresh

weight.

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Carotenoid Profiles: The Paradox of Heat Enhancement

Table 3 shows carotenoid levels and processing impacts in the seven fruit species that

contain quantifiable carotenoids. Analysis of carotenoids demonstrated a mechanistically

important and striking break in the general pattern of loss of phytochemicals during

processing. Thermal pasteurization in tomato had a significant (p < 0.05) positive effect

on total carotenoid extractability (34.8) relative to fresh controls, and on the content of

lycopene ( 38.2). This increase in heat is indicative of the established destabilization of

chromoplast membranes and protein-carotenoid complexes, which release carotenoids out

of their structural relationships and exert more carotenoids into lipid micelles during

digestion (Patras et al., 2023). Pasteurization in mango, papaya, and orange resulted in

small changes in carotenoid content (1222% reduction) instead of the increases in

carotenoids in tomato, indicating that tissue architecture, carotenoid esterification forms,

and cell wall polysaccharide structure of these fruits are not conducive to the same heat-

liberation processes acting in tomato chromoplasts (Oliveira By far, freeze-drying was the

best mode of preservation of carotenoid content (91.3-96.7 percent retention) compared

to the other modalities of drying (conventional hot-air drying gave poor results with

lycopene in tomato with only 61.3 percent retention as a result of isomerization and

oxidative destruction).

Table 3

Carotenoid Content (mg β-Carotene Equivalents/100 g FW) Across Processing

Conditions

Fresh (mg

Fruit

Species

Freeze-

Dried

Pasteurized

Frozen

Key Observation

β-Car.

Eq./100g)

11.3 ±

0.7*†

+34.5% (heat

enhances lycopene)

Tomato

8.4 ± 0.5

7.9 ± 0.5 8.1 ± 0.5

16.7 ±

1.0

13.0 ±

0.8*

15.2 ±

0.9*

16.3 ±

1.0

−22.2% pasteurization

Mango

Papaya

loss

27.3 ±

22.4 ±

24.9 ±

26.8 ±

−17.9% pasteurization

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Fresh (mg

Fruit

Species

Freeze-

Dried

Pasteurized

Frozen

Key Observation

β-Car.

Eq./100g)

1.6

1.3*

1.5*

1.6

loss

4.7 ±

0.4*

−21.6% pasteurization

Orange

5.1 ± 0.4

4.0 ± 0.3*

5.0 ± 0.4

loss

−18.8% pasteurization

Pineapple

Blueberry

Strawberry

3.2 ± 0.3

1.8 ± 0.2

1.2 ± 0.1

2.6 ± 0.2* 3.0 ± 0.3 3.1 ± 0.3

1.4 ± 0.1* 1.7 ± 0.2 1.8 ± 0.2

0.9 ± 0.1* 1.1 ± 0.1 1.2 ± 0.1

loss

−22.2% pasteurization

loss

−25.0% pasteurization

loss

Note. Values are means ± SD (n = 15). * indicates significant difference from fresh

control (p < 0.05). † indicates value significantly higher than fresh control (p < 0.05).

β-Car. Eq. = beta-carotene equivalents. FW = fresh weight. Past. = pasteurized (75°C,

20 min).

Antioxidant Activity: Correlations and Processing Effects

Table 4 shows results of DPPH and FRAP antioxidant activity in various species and

processing conditions. Fresh pomegranate had the best antioxidant activity in all three

assays (DPPH: 18.4 ± 0.9 mmol TE/100 g FW; FRAP: 21.7 ± 1.1 mmol Fe 2+/100

g FW; ABTS: 19.8 ± 1.0 mmol TE/100 g FW TPC showed strong positive relationships

with all the antioxidant measures (DPPH: r = 0.91; FRAP: r = 0.89; ABTS: r = 0.93; all

p < 0.001) which proves that total polyphenol content is the major determinant of

antioxidant capacity in fruit matrices, as demonstrated by Porrini et al. (20

The loss in antioxidant activity due to thermal pasteurization resembling TPC losses per

species with a mean loss of 24.8 (range 14.2 to 41.6) across all of the assays. The

surprising result was that the FRAP activity of thermally pasteurized tomato samples was

much higher than fresh controls due to the higher extractability of heat-stable carotenoids

and phenolic acids with a high ferric reducing capacity. This point shows how

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mechanistically complex heat effects on antioxidant matrices are, and how multi-assays

can be used to characterize them in detail (Huang et al., 2024).

Table 4

DPPH Radical Scavenging Activity and FRAP (mmol TE or Fe²⁺ eq./100 g FW)

Across Processing Conditions

DPPH

Fresh

DPPH

Past.

DPPH

Frozen

FRAP

Fresh

FRAP

Past.

FRAP

Frozen

Fruit Species

Pomegranate

Blueberry

Grape

18.4 ±

0.9

13.1 ±

0.6*

16.7 ±

0.8*

21.7 ±

1.1

15.3 ±

0.7*

20.1 ±

1.0*

14.2 ±

0.7

9.8 ±

0.5*

12.9 ±

0.6*

16.8 ±

0.8

11.4 ±

0.6*

15.4 ±

0.8*

12.8 ±

0.6

8.4 ±

0.4*

11.5 ±

0.6*

15.1 ±

0.7

9.8 ±

0.5*

13.9 ±

0.7*

10.1 ±

0.5

5.9 ±

0.3*

9.0 ±

0.4*

11.8 ±

0.6

6.9 ±

0.3*

10.8 ±

0.5*

Strawberry

Apple

5.3 ±

0.3*

6.7 ±

0.3*

6.2 ±

0.3*

7.9 ±

0.4*

7.4 ± 0.4

6.1 ± 0.3

4.8 ± 0.3

4.3 ± 0.2

3.9 ± 0.2

2.1 ± 0.2

8.6 ± 0.4

7.2 ± 0.4

5.6 ± 0.3

5.1 ± 0.3

4.6 ± 0.2

2.5 ± 0.2

4.2 ±

0.2*

5.5 ±

0.3*

4.9 ±

0.3*

6.6 ±

0.3*

Mango

5.4 ±

0.3*†

4.4 ±

0.2*

6.3 ±

0.3*†

5.1 ±

0.3*

Tomato

Orange

3.0 ±

0.2*

3.9 ±

0.2*

3.5 ±

0.2*

4.6 ±

0.2*

2.6 ±

0.2*

3.5 ±

0.2*

3.0 ±

0.2*

4.2 ±

0.2*

Papaya

1.6 ±

0.1*

1.9 ±

0.2*

1.9 ±

0.1*

2.3 ±

0.2*

Pineapple

Note. Values are means ± SD (n = 15). * indicates significant difference from fresh

control (p < 0.05). † indicates value significantly higher than fresh control. DPPH and

FRAP values expressed as mmol Trolox equivalents (TE) and mmol Fe²⁺ equivalents

per 100 g FW, respectively. Past. = pasteurized.

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Summary of Phytochemical Retention Across Processing Modalities

Table 5 presents a summarized maintenance of mean phytochemical and antioxidant

activity retention percentages in all ten fruit species under each processing condition. The

data prove that freezing and freeze-drying are always better than thermal pasteurization in

all the parameters of phytochemicals except carotenoids in tomato. Ascorbic acid is most

susceptible to thermal processing (3470% retained following pasteurization) and total

phenolics and carotenoids are less susceptible. The consistently high retention values of

freeze-drying (88-97% depending on parameters) place this technology as the ideal

method of preservation of high-quality phytochemical-enriched fruit products despite the

increased cost of production compared to conventional thermal processing.

Table 5

Summary of Mean Phytochemical and Antioxidant Activity Retention (%) Relative to

Fresh Controls Across All Ten Fruit Species

Pasteurized

Retention

(%)

Frozen

Retention

(%)

Freeze-Dried

Retention

(%)

Phytochemical

Parameter

Key Finding

Total Phenolic

Content

Freezing & freeze-

drying superior

57–81%

85–93%

91–96%

Total Flavonoid

Content

Anthocyanins most

sensitive to heat

54–78%

83–91%

89–95%

Extreme

thermolability in all

species

Ascorbic Acid

Anthocyanins

Carotenoids

34–70%

49–65%

78–88%

58–84%

76–92%

88–93%

84–92%

87–93%

89–97%

88–96%

91–97%

90–96%

Strawberry most

susceptible (51.2%

loss)

Tomato exception:

+34.5% after

pasteurization

Correlates strongly

with TPC (r =

0.91)

DPPH Activity

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Pasteurized

Retention

(%)

Frozen

Retention

(%)

Freeze-Dried

Retention

(%)

Phytochemical

Parameter

Key Finding

Tomato FRAP

elevated by heat

(+13%)

FRAP Activity

55–81%

86–93%

89–96%

Note. Retention values represent ranges of species means (n = 10 fruit species per

processing condition). Tomato is excluded from carotenoid pasteurization retention

calculation due to its anomalous heat-enhancement effect. DPPH and FRAP represent

ranges across all species except where noted.

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Discussion

Principal Component Analysis and Multivariate Interpretation

Principal Component Analysis of the entire data set (10 species x 4 processing conditions

x 7 phytochemical/antioxidant parameters) showed that the first two principal

components accounted 71.4 percent total data variance (PC1: 52.3; PC2: 19.1). TPC,

TFC, anthocyanin content, and antioxidant activity measures all strongly loaded PC1 and

effectively, represent a total polyphenol-antioxidant axis. Ascorbic acid and carotenoid

content positively loaded PC2; these two phytochemical classes show an inverse

relationship, which is often observed. Condition clustering Processing confirmed that

fresh and freeze-dried samples were overlapping at positive PC1 values, whereas thermally

pasteurized samples were consistently pushed down to lower PC1 values, and systematic

polyphenol-antioxidant degradation was observed. A unique high location of thermally

pasteurized tomato on PC2 was in line with increased content of carotenoids (Ramos et

al., 2023).

Implications to Public Health and Practices in the Food Industry

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The general conclusions have a number of significant implications on nutritional policy,

consumer advice, and food production practice. The high losses of phytochemicals caused

by thermal pasteurization, especially anthocyanins and ascorbic acid, imply that the

existing nutrient labeling standards can greatly overestimate the phytochemical

composition of thermally processed foods. To the consumers, the data are a strong

indication that fresh or frozen fruits should be of higher priority than thermally processed

alternatives of the species where anthocyanin and ascorbic acid are of nutritional

importance, especially berries and citrus fruits. This nutritional similarity of fresh and well

frozen fruits offers scientific basis to mass health communications that de-stigmatize the

use of frozen fruits (Barba et al., 2023). The paradoxical result that thermal processing

greatly increases lycopene levels in tomatoes has significant practical implications, since

lycopene is a clinically proven antioxidant with proven links to decreased prostate cancer

risk. The message on the health of the people should convey that tomato products that

are heat-processed are the best dietary sources of bioavailable lycopene as opposed to fresh

tomatoes that are eaten without heat processing. In the case of the food industry, the

optimization of thermal processing conditions, including adoption of HTST

pasteurization procedures, minimization of temperatures and times in line with food safety

standards, and adoption of oxygen-exclusion packaging, could significantly decrease

phytochemical losses and preserve the safety of the product (Wojdyło et al., 2024).

Future Perspectives (Integrated with Current Study and Supporting Literature)

Building upon the current findings—particularly the superior phytochemical retention

observed in freezing and freeze-drying, alongside the compound-specific responses to

thermal processing—future research should adopt a systems-level, interdisciplinary

approach that integrates food processing, nutrition, microbiology, and sustainability

science.

A key limitation of the present work is its reliance on in vitro antioxidant assays, which,

although strongly correlated with total phenolic content and antioxidant activity, do not

fully capture physiological relevance. Future studies should extend toward clinical and

metabolic validation, aligning with emerging evidence that functional foods exert

measurable health outcomes. For instance, the demonstrated role of probiotic-enriched

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foods in metabolic regulation and weight management (Rashid et al., 2026) and the

modulation of epigenetic markers by phytochemical-rich diets (Butt et al., 2026b) suggest

that processing-induced changes in phytochemicals may directly influence gene expression

and metabolic health pathways. This creates a direct conceptual bridge between the

antioxidant findings of this study and real-world health outcomes.

Moreover, the observed variability in phytochemical stability—such as the degradation of

ascorbic acid versus enhanced carotenoid bioavailability in tomato—calls for mechanistic

and molecular-level investigations. This aligns with prior work on nutritional bioefficacy

and safety evaluation of novel protein systems (Butt et al., 2025a), where biological

responses to food matrices were shown to depend on structural and biochemical

interactions. Future studies should therefore employ omics technologies (metabolomics,

nutrigenomics) to decode how processing modifies food matrices and downstream

biological effects.

The integration of functional ingredients and microbial systems represents another

promising direction. The use of Lactobacillus rhamnosus in fermented products (Ahmed

et al., 2024) highlights how fermentation can enhance bioavailability, stability, and

functional properties of nutrients. This is particularly relevant to fruit systems, where

controlled fermentation could mitigate phytochemical losses observed during thermal

processing, thereby linking microbial biotechnology with the preservation trends

identified in this study.

From a food systems perspective, the comparative findings on processing methods should

be extended into product development and reformulation strategies. Research on hybrid

protein systems and sustainable formulations (Butt et al., 2025b) demonstrates how

nutritional optimization can be combined with environmental considerations. Similarly,

comparative analyses of food quality and safety (Butt et al., 2024; Butt et al., 2025c)

reinforce the need to balance nutritional retention, microbial safety, and sensory

acceptability—a triad that future fruit processing innovations must address.

In addition, the strong correlation between phytochemicals and antioxidant activity

observed in this study should be contextualized within broader dietary patterns and health

interventions. Evidence from hepatoprotective dietary lipid studies (Khan et al., 2024)

and micronutrient regulation of growth factors (Butt et al., 2026a) indicates that

nutritional outcomes are rarely driven by single compounds, but rather by synergistic

dietary interactions. This reinforces the importance of studying whole-food systems rather

than isolated phytochemicals, supporting the holistic interpretation already discussed in

the manuscript .

Technologically, future research should prioritize next-generation processing techniques

such as pulsed electric fields, cold plasma, and AI-optimized processing systems. The

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growing role of artificial intelligence in optimizing complex systems (Kamal & Butt, 2026)

suggests that predictive modeling could be applied to forecast phytochemical degradation

kinetics and optimize processing parameters in real time, extending the predictive

framework already introduced in this study.

Finally, sustainability must be embedded as a central pillar of future work. While freeze-

drying showed optimal phytochemical retention, its energy intensity raises concerns.

Insights from sustainability-focused research (Khurshid et al., 2026) emphasize the

importance of integrating environmental, social, and economic dimensions into food

processing decisions. Future studies should therefore conduct life cycle assessments (LCA)

of processing techniques, ensuring that nutritional benefits do not come at

disproportionate environmental costs.

Even broader interdisciplinary connections—such as biomechanical and physiological

adaptations in human systems (Mahmood et al., 2026)—highlight an emerging paradigm:

nutrition, processing, and human performance are deeply interconnected, and should be

studied within unified frameworks rather than isolated domains.

In synthesis, future research should aim to:

• Bridge processing–nutrition–health outcomes through clinical and nutrigenomic

studies

• Integrate microbial, functional, and hybrid food systems to enhance phytochemical

stability

• Apply AI-driven predictive modeling for process optimization

• Balance nutritional quality with sustainability and scalability

• Transition from compound-level to system-level understanding of food

functionality

Such an integrated direction not only strengthens the implications of the present findings

but also positions fruit processing research within the broader landscape of functional

foods, precision nutrition, and sustainable food systems development.

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Conclusion

This study has presented a multi-species, multi-modes comparative study of

phytochemical composition and antioxidant activity in fresh and processed fruits, which

have provided data that significantly add to our mechanistic knowledge of the effects of

processing on phytochemical changes. These main findings, which are systematically

recorded in five data tables, attest to the fact that: (1) thermal pasteurization reliably leads

to lowering of TPC, TFC, anthocyanin content and ascorbic acid in all fruit species

studied, with decreases of 18-65% depending on species and compound type; (2) freeze

preserve. The overall implications of these findings are that the specificity of the

processing method used should be taken into account when giving dietary guidance and

nutrient labeling, and that non-thermal and slightly thermal types of processing should be

further invested in. Future studies must focus on investigating phytochemical bio

accessibility and bioavailability in processed fruits-not only in vitro extractability to in

vivo absorption, metabolism, and biological efficacy but also in vivo using sophisticated

analytical methods such as untargeted metabolomics to capture the entire scope of

phytochemical processing changes in a multitude of fruit substrates.

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