Luận văn thạc sĩ VNUA effect of maturity stage and harvest location on chemical compositon and antioxidant capacity of extracts from different parts of musa balbisiana colla fruit

VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE NGO THI HUYEN TRANG EFFECT OF MATURITY STAGE AND HARVEST LOCATION ON CHEMICAL COMPOSITON AND ANTIOXIDANT CAPACITY OF EXTRACTS FROM DIFFERENT

Introduction

Start of art

Growing interest in foods containing natural compounds that support health has emerged as a new research trend, drawing attention from scientists Plants harbor valuable secondary metabolites, including triterpenoids, carotenoids, alkaloids, and phenolic compounds, many of which are polyphenols A substantial body of research demonstrates a link between human health and the intake of polyphenol-rich foods, highlighting the potential of plant-based foods to promote wellness through bioactive polyphenols.

Phenolic compounds are well known for their antioxidant properties and their potential to aid cancer prevention, reduce cardiovascular disease risk, extend lifespan, and help prevent and manage several chronic diseases Research suggests that polyphenol intake can lower diabetes risk and slow aging, while phenolic compounds may also support brain health by mitigating neurodegeneration through reductions in oxidative stress and chronic inflammation, as noted by Williamson (2005) and Cicerale (2012).

Polyphenols are key coloring substances in plants, protecting them from UV radiation, microbial invasion, and insects In plant-based foods, they play a primary role in color, taste, and aroma Polyphenolic compounds are classified into phenolic acids, stilbenes, flavonoids, lignans, and lignins Among these, stilbenes are especially noted for biological activities such as antioxidant effects, prevention of cancer and cardiovascular disease, and anti-inflammatory properties Consequently, stilbene compounds have attracted substantial attention from researchers in recent years.

Besides resveratrol, a stilbene commonly associated with red wine, piceatannol has recently attracted substantial scientific interest Compared with resveratrol, piceatannol exhibits higher biological activity due to its additional hydroxyl group in its structure Research into stilbenes in general, and into resveratrol and piceatannol in particular, from natural sources is increasingly important for understanding their potential benefits and applications.

“Chuoi hot” (Musa balbisiana Colla) has been utilized in Vietnamese traditional medicine for a long time Every part of it is used for curing deseases

Bananas of Musa balbisiana Colla are eaten ripe like regular bananas and are used to treat digestive ailments, while the green fruits and the seeds of ripe fruits are used to treat diabetes and kidney stone disease Historically, the application of Musa balbisiana Colla in traditional medicine has relied mostly on folk experience, with limited scientific explanation and little information about its chemical composition Musa balbisiana Colla contains flavonoids (leuco-anthocyanins), coumarins, tannins, phytosterols (β-sitosterol), and stilbenes, which are strong antioxidants (Huynh et al., 2002) Our unpublished research shows that Musa balbisiana Colla contains a high content of piceatannol.

Therefore, Musa balbisiana Colla may be a new source of piceatannol as well as of other phenolic compounds for application in food and drug industries

Studies indicate that the maturation stage influences the accumulation and profile of antioxidant polyphenols in bananas For example, ripe bananas have lower tannin content but higher anthocyanin levels than green ones, while environmental conditions such as light intensity, soil type, and nutrient supply also affect secondary metabolite accumulation in the fruit To generate scientific data on the phenolic antioxidant content of seeded Musa balbisiana Colla and to identify the maturity stage that yields the richest polyphenols—especially the stilbene piceatannol—we examine the effect of maturity stage and harvest location on the chemical composition and antioxidant capacity of extracts from different parts of Musa balbisiana Colla fruit.

Objectives

This study evaluates the effects of maturity stage and harvest location on the physical-chemical properties, total polyphenol content, piceatannol content, and antioxidant capacity of Musa balbisiana Colla, providing insights into how these factors influence banana bioactivity The results identify the optimal harvest timing and conditions to maximize biological activity by enhancing polyphenol profiles and antioxidant capacity.

This study examines the effect of maturity stage and harvest location on the fruit’s physical-chemical properties, with emphasis on the mass percentage of each component—peel, flesh, and seed—and on the flesh’s hardness and sugar content By comparing fruits harvested at different maturity levels from multiple locations, the work seeks to quantify how component distribution, texture, and sweetness are shaped by maturation and origin, providing insights into fruit quality and its suitability for processing or consumption.

To determine the effect of maturity stage on banana chemistry, this study measures total polyphenol content and antioxidant capacity in the banana’s various parts across different maturity stages It also quantifies piceatannol content in the seeds at each stage of maturity.

Literature review

Characteristics and classification

“Chuoi hot” (seedy banana) has latin name of Musa balbisiana Colla and belonge to Musa genus, Musaceae family, Scitaminae class (Borborah et al.,

Musa balbisiana Colla is a herbaceous plant with big root (banana root)

On the upper stem, a bundle of large, succulent leaves wraps tightly in layers The plant reaches a height of 2–4 meters Each leaf measures 1–1.5 meters in length and sits on a stout, spear-shaped stalk, with a prominent central vein that is convex on the underside and flanked by parallel secondary veins.

The fruit is succulent and big with 5 edges, contains 4-5 mm black ball-shaped seeds whose embryo is white (Pham Hoang To, 2014)

Figure 1 Trees, branch fruit and seeds of “ chuoi hot” source : http://data.abuledu.org/wp/?LOM024 and http://www.bananas.org

Most of consumed banana varieties are hybridizations of 2 wild species called Musa acuminata Colla and Musa balbisiana Colla (Stover and Simmonds,

1987) The differences between these 2 species are listed in the Table 2.1 Table2.1 Characters used in the clasiffication of banana though a taxonomic scorecard

Character Musa acuminata Musa balbisiana

Pseudostem color More or less heavily marked with brown or black blotches

Petiolar canal Margin erect or spreading, with scarious wing below, not clasping pseudostem

Margin inclosed, not winged below, clasping pseudostem

Peduncle Usually downy or hairy Glabrous

Ovules Two regular rows in each loculus

Four irregular rows in each loculus

Bract shoulder Usually high ( ratio 0.30) Bract curling Bract reflex and roll back Bracts lift but no roll

Bract shape Lanceolate or narrowly ovate, tapering sharply from the shoulder

Broadly ovate, not tapering sharply

Bract color Red, dull purple or yellow outside; pink, dull purple or yellow inside

Distinctive brownish-purple outside; bright crimson inside

Color fading Inside bract color fades to yellow towards the base

Inside bract color continuos to base

Bract scars Prominent Scarcely prominent

Free tepal of male flower Variably corrugated below tip

Male flower color Creamy white Variably flushed with pink

Stigma color Orange or rich yellow Cream, pale yellow or pale pink Source: Simmonds and Shepherd (1955)

Musa balbisiana Colla grows primarily in Southeast Asia and the southern part of China In Vietnam, it grows in the northern mountainous provinces, including Yen Bai, Lao Cai, Lang Son, and Hoa Binh.

Musa balbisiana Colla is a hydrophyte with greater vitality than other species, offering notable shade tolerance and competitive vigor that makes it useful for land protection and landscape planting For practical landscaping, people often place it in garden corners, under the shade of fruit trees, or beside bamboo It propagates efficiently: each year 1 to 3 new plants can arise from a single mother stem, and its seeds also germinate readily (Pham Hoang To, 2014).

2.1.3 Nutritious compositon and bioactive compounds

Bananas deliver a mix of carbohydrates, minerals, protein, fiber, and essential vitamins, making them a valuable part of the human diet They contain all 10 essential amino acids, supporting growth and health, which is particularly beneficial for children and older adults The banana’s carbohydrate content changes markedly as it ripens, affecting energy availability In addition to energy, bananas supply important nutrients—potassium, vitamin B6, and vitamin C—along with fatty acids such as palmitic, linoleic, alpha‑linolenic, and oleic acid that contribute to overall health and energy metabolism Both the flesh and the peel contain beta-carotene, with concentrations ranging from about 40 to 4,960 µg per 100 g (Mohapatra et al., 2010; Pothavorn et al., 2010).

Table 2.2: Nutritious composition in banana flesh

Component Without peel (g/100g banana powder )

Bananas contain powerful antioxidants and several bioactive amines, including serotonin, norepinephrine, dopamine, and other catecholamines Dopamine is a key brain neurotransmitter, and in ripe bananas (chuoi hot) it may act as an antioxidant, with reported concentrations ranging from 80–560 mg per 100 g and 2.5–100 mg in fresh fruit, according to Emaga et al (2008b) and Kanazawa and Sakakibara (2000).

Banana flesh and peel contain notable bioactive compounds, including flavonol glycosides such as rutin (approximately 242.2–618.7 mg/g of dry weight) and antioxidant tannins, which have been linked to various health benefits (Mohapatra et al., 2010; Tsamo et al., 2014) In addition, research has reported the presence of leucocyanidin in banana flesh, a compound associated with anti-ulcerogenic effects and potential protection against gastric ulcers (Lewis et al., 1999).

Studies on different parts of Musa balbisiana Colla fruits show that the bracts contain anthocyanins, with delphinidin and cyanidin identified as the two main anthocyanins (Horry and Ray, 1987) Japanese researchers indicate that some phytoalexins include 1,2,3,4-tetrahydro-6,7-dihydroxy-1-(4’-hydroxycinnamyliden)naphthalen-2-one and 2-(4’-methoxyphenyl).

- 1,8 - naphthalic anhydrid; 2 - phenyl - 1,8 - naphthalic anhydride are present in the banana fruits (Kamo et al., 1998)

Three neo-clerodane diterpenoids, Musa balbisian A, Musa balbisian B, and Musa balbisian C, were reported from Musa balbisiana seeds (Ali, 1991) At Ho Chi Minh City University of Medicine and Pharmacy, Nguyen Thi My Hanh and Bui My Linh analyzed the chemical composition of chuoi hot seeds and found saponins, coumarins, tannins, flavonoids, anthocyanosides, uronic compounds, essential oils, and phytosterols The study relied on qualitative tests, and the identification and quantification of individual constituents were not performed.

Resins from Musa babisiana contain caffeoylquinic acid, myricetin-3-O-rutinoside, and a myricetin glycoside, indicating the presence of several phenolic compounds in this banana species Additional banana phenolics identified include dopamine, N-acetylserotonin, kaempferol-3-O-rutinoside, quercetin-3-O-rutinoside, naringenin glycosides, and an apigenin glycoside I, with naringenin glycoside II detected by absorption spectral analysis in the 280–320 nm range (Pothavorn et al., 2010).

In Thailand, 6 anthocyanins are identified in the banana flowers by HPLC-

MS method: delphinidin-3-rutinoside, cyanidin-3-rutinoside, petunidin-3- rutinoside, pelargonidin-3-rutinoside, peonidin-3-rutinoside, and malvidin- 3- rutinoside Musa babisiana fruit contains delphinidin-3-rutinoside and cyanidin- 3-rutinoside (Kitdamrongsont et al., 2008)

A study of the flesh and peel of 13 banana varieties identified numerous phenolic compounds, including caffeic acid-hexoside, ferulic acid-hexoside, sinapic acid-hexoside, ferulic acid-dihexoside, myricetin-deoxyhexose-hexoside (high in pulp), quercetin-deoxyhexose-hexoside (high in pulp), methymycricetin-deoxyhexose-hexoside, quercetin-hexoside, and isorhamnetin-3-O-rutinoside (Tsamo et al., 2015).

In Spain, a research group separated compounds from the chloroform extract of “chuoi hot,” including a fatty ester of phytol, a fatty ester of an n-alkanol, beta-sitosterol, and stigmasta-5,22-dien-3β-ol From the acetone extract, they isolated a (+)-epiafzelechin compound The epiafzelechin was tested for its ability to prevent Cryptolestes pusillus Schoenherr, an insect harmful to cereals (Pascual-Villalobos and Rodríguez, 2007).

Research on the chemical composition of three Musa species at three ripeness levels reveals that their flesh contains alkaloids, saponins, glycosides, flavonoids and tannins, with the concentrations of these compounds varying across ripeness stages (Obiageli A et al., 2016).

Researchers at the National Science and Technology Center conducted a preliminary study of the composition of chuối hột in Vietnam, identifying two compounds: cyclomusalenon, (24S)-24-methyl-29-norcycloart-25-en-3-one, and stigmasterol Stigmasterol is a very common plant sterol found in nature, while cyclomusalenon is a five-ring triterpenoid featuring a cyclopropane ring and a 3-oxo-29-norcycloart skeleton, a structure rarely observed in nature (Tran et al., 2003).

The National Science and Technology Center, in collaboration with Hanoi Medical University, studied the blood glucose–lowering effects of “chuoi hot” in an in vivo mouse model, administering the extract by intradermal injection They found that chuoi hot extract produced superior hypoglycemic activity compared with the root extract of Anemarrhena asphodeloides Bunge and was comparable to the root extract of Smilax glabra Roxb at the same concentration Notably, cyclomusalenone accounts for about 0.85% of the extract but its hypoglycemic effect is nearly equivalent to 0.82% of the total The hypoglycemic activity of chuoi hot appears to be attributed to docyclomusalenone, according to Q V et al.

Like other kinds of fruit, unripe Musa balbisiana tastes extremely astringent

Riper bananas exhibit a milder, less astringent taste than their unripe counterparts Unripe Musa balbisiana has historically been used to treat conditions such as cystolith, ringworm, and back pain Our unpublished recent research identifies piceatannol in banana seeds, suggesting that Musa balbisiana contains polyphenol compounds However, to date no publication has reported the phenolic composition of Musa balbisiana.

2.1.4 Uses of “chuoi hot” in Vietnam

Phenolic compounds

Phenolic compounds are aromatic molecules with hydroxyl groups directly attached to a benzene ring Molecules bearing multiple hydroxyl groups on a single benzene ring are called polyhydroxyphenols (monomers); when many such monomers link together, they form polymers (Le Ngoc Tu, 2003).

Polyphenols show a wide diversity in their structures, functions, and distribution across plants, which leads to multiple classification schemes They can be grouped by origin, biological roles, and chemical structures Because the structure of phenolic compounds is linked to their biosynthetic pathways in the carbon cycle, phenolics are categorized into distinct groups, as illustrated in Figure 2.3.

Figure 2.3 Classification and structure of major phenolic compound

Source: Adapted from Han et al (2007)

Phenolic acids are abundant in plants, and they are classified by structural features into two main groups: hydroxybenzoic acids and hydroxycinnamic acids Hydroxybenzoic acids are based on a benzene ring with hydroxyl substitutions and do not possess a carbon side chain, while hydroxycinnamic acids contain a three-carbon side chain attached to the ring, forming the cinnamic acid framework.

Hydroxycinnamic acids contain many hydroxyl and methyl groups in the struture

They are materials for lignin synthesis and many other compounds

Hydroxybensoic acids are found with low content in edible plants In plants, they are the raw material for the synthesis of lignin and hydrolysis tannin

 Flavanoid Flavonoid is a secondary metabolite product of plants, with a carbon chain of

Flavonoids share a C6-C3-C6 carbon skeleton, and their classification depends on features of this chain, notably the pattern of double bonds and the presence of hydroxyl groups Based on these characteristics, flavonoids are grouped into flavonols, flavanols, flavones, isoflavones, flavanones, and anthocyanins These compounds are abundant in plants, a point noted by Robards and Antolovic.

1997) They have strong antioxidant capacity Besides, some flavonoids have anti-inflammatory, anti-allergy, anti-inflammatory, antibacterial properties

Lignin is a distinctive plant polymeric compound found in many wood tissues, acting as a cell adhesive that increases mechanical strength, waterproofs the xylem cell wall, and helps prevent infiltration by pathogenic microorganisms It is the condensation product of phenylpropane units, with two phenylpropane units coalescing to form lignan Lignin is abundant in linseed, reaching up to 3.7 g/kg dry matter Lignin and its derivatives are of interest in research because they are thought to be feasible for use in the treatment of cancer and other diseases (Salee, 2005).

 Tanin Tanin is a mixture of C6 - C1 and C6 - C1 - C6 (gallic acid and diagallic acid in free form and glucose - conjugated form) Tanin compounds are common in plants and classified into 2 types:

- Condensed tanin Tannins are popular in some trees such as guava, banana, persimmons, etc , Tannin content is very different in different parts of the plant

 Stilbene Stilbene is a small molecular weight compound (MW = 210 ÷ 270), which is a natural secondary compound that protects plants against bacteria, preventing bad effects from ultraviolet light and some serious diseases Stilbene is synthesized via the phenylpropanoid route The synthesis of stilbene synthesized by plants mainly depends on the stimulation of the environment The five most common stilbene compounds in nature include: resveratrol, piceatannol, pinosylvin, rhapontigenin and pterostilbene (Roupe et al., 2006) Among these give stilbenes, resveratrol and piceatannol are well studied by researchers

Piceatannol(3,5,3',4'-tetrahydroxystilbene;5-[2-(3,4dihydroxyphenyl) ethenyl] benzene-1,3-diol is derirative of to resveratrol

Figure 3.4 Structure of Piceatannol and Resveratrol

Piceatanol is has molecular formula of C14H12O4 Piceatannol is a white powder, has a melting point at 223 0 C-226 0 C, molecular weight of 244.24 This compound is insoluble in water but soluble in ethanol and dimethyl sulphoxide

Spectral analysis of piceatannol in ethanol showed that piceatannol absorbed up to 322 nm, while trans-resveratrol absorbed maximum at 308 nm (Rossi et al,

Piceatanol is proven to be highly bioactive, anti-oxidant, anti-inflammatory, anti-obesity and diabetes, anti-cancer, cardiovascular (Piotrowska et al, 2012)

Using foods containing high resveratrol and piceatannol helps to reduce the risk of cardiovascular disease, to prolong longevity and to enhance human health (Roup et al., 2006)

Piceatannol is found in many plants, with red wine and grapes being the most important sources in the human diet, though its concentration is lower than that of resveratrol in grapes In grapes, piceatannol is 0.78 μg/g and resveratrol 3.18 μg/g, whereas in red wine piceatannol is 908 μg/g and resveratrol 208 μg/g, making piceatannol more abundant in red wine (Cantos et al., 2000).

According to our present study, the piceatanol content in the sim is 2.3 mg/g dry matter, 1000-2000 times greater than the one of red grape (Lai et al., 2013)

In addition, piceatannol is also found in lemon creeper, Asian beans, peanut, and so on

2.2.2 Biological activity of phenolic compound

 Antioxidant capacity Antioxidant activity is the most studied property of phenolic compounds

Antioxidants, particularly phenolic compounds, slow or inhibit the oxidative processes driven by excessive reactive oxygen species (ROS) and reactive nitrogen species (RNS), helping to mitigate oxidative stress and protect cellular integrity.

Reactive oxygen and nitrogen species (ROS and RNS) have dual roles in biology: they are necessary at low to moderate levels for normal cellular function and defense against pathogens, yet they can be damaging when produced in excess Their levels are kept in check by endogenous antioxidants, including enzymatic systems and antioxidant vitamins such as vitamins E and C But exposure to agents like ionising radiation, ultraviolet light, tobacco smoke, ozone, and nitrogen oxides in polluted air can trigger oxidative stress, defined by an excess of ROS and RNS alongside depleted antioxidant defenses When ROS and RNS exceed the cell’s antioxidant capacity, they can damage lipids, proteins, and DNA, impairing their normal functions.

Phenolic compounds are potent dietary antioxidants that bolster the body's antioxidant defense, working in concert with carotenoids and antioxidant vitamins to counteract oxidative stress The antioxidant actions of phenolics are well established and include direct scavenging of free radicals, chelation of transition metal ions, and inhibition of enzymes such as xanthine oxidase that generate radicals.

Cardiovascular diseases are the leading cause of death in the United States, Europe, and Japan, and accumulating evidence implicates oxidative stress in driving major cardiovascular dysfunction Increased ROS production contributes to atherosclerosis through four main mechanisms: oxidation of LDL to oxidised-LDL, endothelial cell dysfunction, smooth muscle cell migration and proliferation with matrix metalloproteinase release, and monocyte adhesion and foam cell formation from oxLDL uptake Phenolic compounds in fruits, cocoa powder and dark chocolate, and coffee have been reported to inhibit LDL oxidation and lower cardiovascular risk Green tea consumption reduces total and LDL cholesterol and decreases LDL oxidizability, correlating with reduced risks of stroke and myocardial infarction Resveratrol and piceatannol, two stilbenes found in red wine, exhibit cardioprotective activities including inhibition of LDL oxidation, support of cardiac cell function, suppression of platelet aggregation, and attenuation of myocardial tissue damage during ischemia; moderate red wine rich in these stilbenes has been linked to the “French Paradox” noted by Renaud and De Lorgeril in 1992.

 Anti-inflammatory activity Inflammation is a dynamic process that is elicited in response to mechanical injuries, burns, microbial infection and other noxious stimuli (Shah et al., 2011)

Inflammation presents with redness, heat, swelling, loss of function, and pain Redness and heat reflect increased blood flow; swelling results from enhanced vascular permeability; pain arises from the activation and sensitization of primary afferent nerve fibers A vast array of inflammatory mediators—including kinins, platelet-activating factor, prostaglandins, leukotrienes, amines, purines, cytokines, chemokines, and adhesion molecules—act on specific targets to provoke local mediator release from leukocytes and to recruit more leukocytes, especially neutrophils, to the site of injury Under normal conditions these responses help contain the insult and protect the organism, but low-grade chronic inflammation is now recognized as a critical factor in many diseases, including cancer, obesity, type II diabetes, cardiovascular and neurodegenerative diseases, and premature aging (Santangelo et al.).

2007) Phenolic compounds have been reported to display marked invitro and invivo antiinflammatory properties via various mechanisms of action including:

The key mechanisms involve inhibiting the arachidonic acid pathway, modulating the nitric oxide synthase family, and regulating the cytokine system along with the NF-kB and MAPK signaling pathways (Santangelo et al., 2007).

Cancer is characterized by two biological properties—uncontrolled cell proliferation and the ability of cancer cells to invade distant sites It results from exposure to a range of carcinogens, including tobacco smoke, alcoholic drinks, industrial toxins, aflatoxins, heterocyclic amines, N-nitroso compounds, and polycyclic aromatic hydrocarbons A wide variety of natural bioactive compounds, including polyphenols, have been shown to inhibit carcinogenesis Phenolic compounds act as anti-cancer agents through multiple mechanisms: their antioxidant properties, modulation of signal transduction pathways, induction of apoptosis, arrest of the cell cycle, and inhibition of cancer cell invasion.

Meterials and methods

Sample and chemical

3.1.1 Sample collection and prepairation Sample collection:

The “Chuoi hot” bananas were harvested in Namdinh and Yenbai provinces for a banana ripening study In each province, three bunches with the same biological maturity were collected From the middle hands of each bunch, at least 30 fruits were gathered and allowed to ripen at room temperature in cardboard The bananas were then separated into five maturation stages, including green, green more than yellow, yellow and green end, and yellow and yellow with brown spots For each ripening stage, three fruits were removed from the cardboard and weighed.

Figure 3.1 Five maturity stages of “chuoi hot”

For freeze dried sample preparation

Each fruit was cut on the length and across the width into four quarters

Two groups were formed by pairing diagonally opposite quarters, with samples from the three fruits pooled in each group In one group, the pulps, peels and seeds were freeze-dried separately and then vacuum-sealed in polypropylene bags The freeze-dried materials were ground into powder, and the resulting powders were stored at −20°C until analysis.

Figure 3.2 All part of “chuoi hot”

Sodium carbonate (Na2CO3); acetone (C3H6O,100%); acetonitrile (C2H3N, 99.8%); 2,2-diphenyl-1-picrylhydrazyl (DPPH); 3,4,5-Trihydroxybenzoic acid monohydrate (gallic acid, monohydrate); Folin-Ciocalteu’s reagent; 6-hydroxyl- 2,5,7,8- tetramethylchroman-2-carboxylic acid ( Trolox)

Electric balance Heat dried oven (Memmet, Germany) Centrifugation (Mikro 220R, Mikro 200R, Hettichzentrifugen, Germany)

Vortex mixer ( JK-VT-F JINGKI SCIENTICIN, China) Speed vacuum (Genevac, England)

Method

Total dry matter was ditermined by drying method to a constant weight at 105 0 C

The stiffness of “chuoi hot” was determined by DIGITAL FIRMNESS TESTER machines in Idian

Figure 3.3 Stiffness machine 3.2.3 Determination sugar profiles

Sample preparation Briefly, 0.3 g of freeze dried sample was weighted into 15ml fancol and mixed with 9ml distilled water by using vortex and then centrifuge 12000 rpm,

4 0 C for 10 min The supernatant was taken for analysis by HPLC equipment

Preparation standard of sugar profiles: A 0.1 g sample of sugar was weighed into a 2 mL microtube, to which 1 mL of distilled water was added and the mixture was thoroughly mixed with a vortex mixer The resulting 10% sugar solution was then diluted to prepare concentrations of 0.25%, 0.5%, 1%, and 1.5%.

Quantification of the sugar profile was performed by high-performance liquid chromatography (HPLC) on a Shimadzu system (Japan) equipped with a DGU-20A3 degasser, LC-10Ai pumps, a CBM-20A Monitor, and an RID detector A 20-µL aliquot of the extract was injected onto a SUPELCOSIL NH2 LC column (25 cm × 4.6 mm, 5-µm particles) with a guard column of the same type (Supelco, Japan) The mobile phase was acetonitrile 80%, with a flow rate of 1.0 mL/min and the column temperature maintained at 30 °C.

Figure 3.4 Chromatography of glucose and fructose at concentration of 0.5%

Figure 3.5 Standard curves of glucose and fructose

3.2.4.Determination total polyphenol content and antioxidant capacity

Phenolic compounds in different parts of chuoi hot were extracted using a protocol previously optimized by our research group Briefly, approximately 0.13 g of freeze-dried sample was mixed with 4 mL of 60% acetone in a water bath and shaken for 60 minutes at 40°C After centrifugation at 6,000 rpm for 10 minutes at 4°C.

The supernatant was collected and evaporated to dryness with a rotary evaporator at 35°C The residue from evaporation was dissolved in 70% methanol and analyzed for total phenolic content, antioxidant capacity, and piceatannol content.

Determination total phenolic content The total phenolic content of the extract was determined by the Folin–Ciocalteu method (Singleton, L and Rossi, 1965)

Briefly, 500 μL of the sample solution diluted to the appropriate concentration was mixed thoroughly with 250 μL of Folin–Ciocalteu reagent (1 N) for 5 minutes, followed by the addition of 1,250 μL of 7.5% Na2CO3 The mixture was allowed to stand for a further 30 minutes in the dark, and absorbance was measured at 755 nm The total phenolic content was calculated from the calibration curve (Figure 3.6) and expressed as mg of gallic acid equivalents per g dry weight (mg GAE/g DW).

Figure 3.6 Gallic standard curve Determination of antioxidant capacity y = 0,0295x + 0,0316 R² = 0,9989

Scavenging activity of DPPH radical was assessed according to the method of Larrauri, Sanchez-Moreno and Saura-Calixto (1998) with some modification

Briefly, 0.1 ml of diluted sample solution was mixed with 2.9 ml of 0.1 mM DPPH methanol solution After the solution was incubated for 30 min at 25

In the DPPH assay, the decrease in absorbance at 517 nm was measured to assess radical scavenging activity The control contained methanol instead of the antioxidant solution, while the blanks contained methanol instead of the DPPH solution The inhibition of DPPH radicals by the sample was calculated using the standard equation for percent inhibition.

Figure 3.7 Trolox standard curve 3.2.5 Determination of piceatannol content

Quantification of the piceatanol was performed by HPLC using a Shimadzu system (Japan) equipped with a DGU-20A3 degasser, LC-10Ai pumps, a CBM-20A Monitor and a SPD-M20A Diode array detector (DAD) A

An aliquot of 20 μL of the extract was injected onto a Phenomenex Kinetex EVO C18 column (150 × 4.6 mm, 5 μm) with a guard column of the same type The mobile phases were A) water with 0.1% formic acid and B) acetonitrile with 0.1% formic acid The flow rate was 1.0 mL/min and the column temperature was 30°C A 42-minute gradient, as shown in Table 3.1, was employed The calibration curve is described by y = 0.0829x − 1.5391 with R² = 0.9982.

Figure 3.8 Chromotogaphy of piceatannol standard at concentration of 100 àg/ml

3.2.6 Statistical analysis Data were analysed using the statistical software Minitab 16.0 Analysis of variance was carried out using a Generalised Linear Model (GLM) procedure to determine the effect of the havest location, maturity stage and their interactions onanalysed index The model conﬁguration was Yi = a + b1*X1 +b2*X2 + b12*X1*X2 (Y: the analysed index; X1: haverst location and X2: maturity stage).Tukey test were used to determine the differences among the means p- values < 0.05 were considered to be significantly different.

Results and discussions

Effects of the maturity stage and harvest location on physical-chemical

4.1.1 The ratio of each part in “chuoi hot”

The propotion of each part in “chuoi hot” fruit were changed during ripening.The result was showed in Figure 4.1

Figure 4.1 Impact of the maturity stage of the “chuoi hot” fruit harvested in

Namdinh and Yenbai on the propotion of different part

Statistical analysis showed that harvest location and maturity stage significantly affected peel percentage (p = 0.000 and p = 0.003, respectively), while the interaction between location and maturity stage was not significant (p = 0.636) The peel proportion in Yen Bai was 34.71 ± 3.96%, substantially higher than in Nam Dinh, where it was 10.04 ± 0.96%—about 3.45 times lower Regarding maturity stage, peel percentage declined markedly from the first to the fifth stage In Nam Dinh, peel decreased gradually from 12.8 ± 0.53% to 7.97 ± 1.34%, a reduction of about 1.6 times from stage 1 to stage 5 In Yen Bai, peel proportion fell from 40.42 ± 2.85% to 31.28 ± 7.35%.

Statistical analysis indicated that harvest location significantly affected the pulp percentage (p = 0.000) Maturity stage and the interaction between harvest location and maturity stage did not significantly affect the pulp percentage (p = 0.051 and p = 0.285, respectively) The average proportion of pulp in Nam Dinh province was reported, though the exact value is not provided in the available data.

Pe rc en ta ge %

Namdinh seed pulp peel a ab ab b ab b b b b b a a a a a

Pe rc en ta ge %

75.13± 1.39% in fruit, while the for the one in Yenbai was lower,with value of 43.16± 5.23%.The percentage of pulp were increased lightly between the 1 st and

Across five maturity stages, pulp content in Nam Dinh province increased modestly from 70.47 ± 1.63% at the 1st maturity stage to 80.10 ± 2.04% at the 5th maturity stage In Yen Bai province, the pulp proportion rose from 40.57 ± 6.53% to 43.42 ± 4.98%, about a 1.07-fold (roughly 7%) increase over the five maturity stages.

Harvest location significantly affected the seed percentage (p = 0.005), while maturity stage and the interaction between harvest location and maturity did not significantly affect seed percentage (p = 0.893 and p = 0.378, respectively) The seed percentage in Yen Bai (22.13 ± 8.62%) was higher than in Nam Dinh (14.83 ± 1.37%) The seed percentage across the five maturity stages was similar at the two harvest locations.

Across two harvest locations, the pulp proportion was highest in the chuoi hot banana, while the peel proportion was lower and the seed proportion the lowest Natural conditions such as climate, temperature, environmental factors, and nutrition can affect fruit composition at different harvest sites Compared with other banana varieties, chuoi hot exhibits a distinct pattern of higher pulp share, illustrating how harvest location and growing conditions shape the overall fruit composition.

Banana varieties chuoi tieu, chuoi su, and chuoi bom have pulp contents of 65%, 72%, and 73%, respectively, while the pulp content of chuoi hot is higher for fruit harvested in Namdinh and lower for fruit harvested in Yenbai, according to Huynh Nguyen Thai Duy (2013).

Hardness is one of the important indicators to evaluate maturiy stage

Green fruit has high hardness and ripen fruit has low hardness

Maturity stage signifficantly effected hardness of pulp (p=0.000) while harvest location, interaction between harvest location and maturity stage did not effect hardness(p = 0.103, p = 0.329)

Among 2 harvest location, hardness of “chuoi hot” harvested in Yenbai was higher than that harvested in Namdinh.The hardness decreased dramatically between the first matuarity to the last matuarity The hardness of “chuoi hot” harvested in Namdinh reduced sharply from 9.32 ± 0.76 kg/cm 2 to 0.489 ± 0.2kg/cm 2 and for Yenbai from 10.31± 1.24kg/cm 2 to 0.39± 0.04kg/cm 2

Figure 4.2 shows a rapid reduction in fruit pulp hardness from the first to the second maturity stage across all harvest locations At Namdinh, hardness decreases from 9.32 ± 0.76 kg/cm2 to 1.92 ± 0.58 kg/cm2, while at Yen Bai it declines from 10.30 ± 1.24 kg/cm2 to 2.70 ± 0.56 kg/cm2, corresponding to approximately 4.84- and 3.81-fold drops, respectively This maturation-related softening is attributed to pectin degradation by pectinase, leading to cell wall loosening and increased water content in the fruit pulp.

In fact that green banana is harder than ripen banana

Namdinh and Yenbai on pulp hardness Many studies showed that hardness was effected by maturity stage

Research on the biochemistry of fruit maturation indicates that firmness declines as fruit ripens: banana firmness decreases by ninefold, mango firmness decreases by ninefold, and papaya firmness decreases by sixtyfold (Bui Quang Huy and Pham Quang Hung, 2009).

Research on quatity assessment of tomato posthravest saw that hardness of tomato decreased gradually with maturity stages (Nguyen Minh Thuy and Nguyen Thi Kim Quyen, 2009)

4.1.3 Changing of sugar content of banana pulp harvest in 2 locations

The result of HPLC analysis shown that the composition of “chuoi hot” pulp of had glucose and fructose a b c c c a bc bc bc 0,0 bc

Fi rm ne ss (k g/ cm 2 )

Figure 4.3: Sugar ptofile of “chuoi hot pulp at 5 th matyrity”

Namdinh and Yenbai on sugar content

Statistical results show that harvest location and maturity stage significantly influence pulp glucose content (p = 0.027 and p < 0.001, respectively) The interaction between harvest location and maturity stage did not have a significant effect on pulp glucose content (p = 0.305) Glucose content in Nam Dinh was higher than in Yen Bai, at 31.24 ± 3.55% versus 26.46 ± 5.55%, respectively Glucose content in the pulp increased from the first to the fifth maturity stage.

As result shown in figure 4.4, glucose content was significantly lower in the first maturity stage than other maturity stages In the 1 st maturity stage the b a a a a b a a a a

Su ga r c on te nt (% D W )

GlucoseFructose glucose content was 2.87 ± 3.93% DW However, the 5 th maturity stage it increased about 13.8 times, reached the value of 39.81 ± 1.63 %DW in Namdinh

“chuoi hot” There was a rapid increase of glucose content in Yenbai from just around 2.00 ± 1.59 %DW to 39.08 ± 1.9 %DW of the 1 st and the 5 th maturity stages, respectively

Statistic results showed that harvest location, matuarity stage sinificantly effected (p=0.044, p=0.000) fructose content in pulp while interaction between harvest location and maturity stage did not have any significant effect (p=0.520)

In Namdinh, the average fructose content of harvested chuoi hot was 22.78% (±3.96%), higher than the 19.15% (±3.9%) recorded for chuoi hot in Yenbai Across all harvest locations, fructose content rose dramatically from the first to the fifth maturity stage.

Sugar content in chuoi hot reaches its peak from the first to the second maturity stage and then stabilizes as the fruit progresses from the second to the fifth maturity stage This sugar pattern helps explain why ripening chuoi hot is sweeter than green fruit Compared with fructose, glucose content is higher in chuoi hot, and the sugar content of chuoi hot harvested in Namdinh is higher than that of chuoi hot harvested in Yenbai.

During fruit maturation, total soluble solids increase as hydrolysis reactions convert complex compounds into simpler ones; starch and tannins decline, forming simple sugars, while lipids also undergo hydrolysis This pattern has been observed in raspberries and strawberries (Wang et al., 2009).

Effect of maturity stage to total polyphenol content of“chuoi hot”

Total polyphenol of different part of “chuoi hot” changed during maturation

Peel: Statistical analysis revealed that the maturity stage significantly influenced the total phenolic content of the peel (P = 0.000), while harvest location did not have a significant effect, and the interaction between harvest location and maturity stage also showed no significant impact on total phenolic content.

Peel samples harvested in Namdinh showed an average total phenolic content of 19.74 ± 2.52 mg GAE/g DW, which was not significantly higher than that of Yenbai at 17.86 ± 5.63 mg GAE/g DW Across the two harvest locations, total polyphenol content declined markedly from the first to the fifth maturity stages: at Namdinh, it decreased from 30.68 ± 1.12 mg GAE/g DW to 11.27 ± 4.51 mg GAE/g DW (a 2.7-fold reduction), and in Yenbai from 34.58 ± 13.61 mg GAE/g DW to 8.94 ± 3.16 mg GAE/g DW (a 3.8-fold drop) Papaya peels showed a gradual fall from 471.97 to 358.67 mg GAE/100 g FW, corresponding to a 1.31-fold decline during ripening (Sancho et al., 2010).

Namdinh and Yenbai on total phenolic content of peel

Statistics analysis result showed that harvest location, maturity stage and interaction between harvest location andmaturity stage significantly effected TTP of pulp (p =0.000, p = 0.000, p = 0.000)

Pulp from Yen Bai showed a higher average total phenolic content than pulp from Nam Dinh, recording 33.15 ± 5.83 mg GAE/g DW versus 10.31 ± 1.31 mg GAE/g DW, respectively Across maturity stages, the 3rd maturity stage exhibited the highest total phenolic content, while the 1st maturity stage had the lowest.

There was a dramatically drop about 2.4 times in total polyphenol content ab abc bc bc c

Po ly ph en ol s (m g G AE /g D W )

Namdinh pulp shows a decrease in total polyphenol content from the 1st maturity stage (19.33 ± 1.78 mg GAE/g DW) to the 2nd stage (8.87 ± 0.78 mg GAE/g DW), with the 2nd–5th maturity stages exhibiting similar polyphenol levels Yenbai pulp shows total phenolic content rising from 11.46 ± 2.82 mg GAE/g DW at the 1st maturity stage to a peak of 52.51 ± 4.55 mg GAE/g DW at the 3rd maturity stage, followed by a reduction in later stages.

5 th (26.2 ± 5.43 mg GAE/g DW) maturity stage was observed Total phenolic content of papaya pulp showed a dramatic decrease from 1.91 to 0.88 mg GAE/100g FW during maturation (Sancho et al., 2010)

Namdinh and Yenbai on total phenolic content of pulp

Harvest location and interaction between harvest location and maturity stage had significant effecton total phenolic content of seed (p =0.000, p = 0.000) while maturity stage did not have any effect (p = 0.595)

The average of total phenolic conten of seed in Yenbai (66.73± 8.71 mg GAE/g DW) was higher the the one in Namdinh (51.93± 5.06 mg GAE/g DW)

Total penolic content of seed in Namdinh fail gradually from the first maturity to the firth maturity By contrast, total phenolic content of seed in Yenbai increased from the 1 st to 5 rd maturity stage.Total polyphenol content of seed in Namdinh had a dramatic decease from 70.19 ± 2.21 mg GAE/g DW to 37.66 ± 6.56 mg GAE/g DW, coresponding to a reduciton of 1.86 time, while total phenolic content of seeds in Yenbai increased with maturity stage from 54.54 ± 11.29 mg bc c c c c

(mg GAE/g DW) to 80.4 ± 5.33 mg (mg GAE/g DW) Total phenolic content of

“chuoi hot” seed was simillar with the one of wild strawberry (Wang et al., 2009)

Namdinh and Yenbai on total phenolic content of seed

Among different parts of “chuoi hot”, seed had the highest total phenolic content, at 59.33 ±14.7 (mg GAE/ g DW), followed by pulp, at 21.73 ± 16.48(mg GAE/g DW) and the peel had the lowest one, at 18.8 ± 9.56 (mg GAE/g DW)

Total phenlic content of seed was higher than the one of pulp and peel of 2.7 and 3.15 times, respectively

Total phenolic content (TPC) in Namdinh decreases gradually during fruit maturation In contrast, TPC in Yen Bai pulp and seeds increases markedly with maturation, with seeds exhibiting the highest TPC and the peel the lowest Environmental factors such as climate, temperature, and water availability can influence total phenolic content, so TPC can differ between these two harvest locations.

Total phenolic content indicates that fruits are the primary source of phenolics, with values ranging from 13 mg GAE/g DW to 29 mg GAE/g DW Specific fruits exhibit the following phenolic levels: Stenocereus stellatus Riccobono: 13.84–15.52 mg GAE/g DW; Malus pumila: 13.00–13.10 mg GAE/g DW; Fragaria ananassa: 16.00–18.00 mg GAE/g DW; Rubus idaeus: 27.00–29.00 mg GAE/g DW; Vaccinium oxycoccus: 22.00 mg GAE/g DW (Carmen, 2009).

Compare to the result of M Carmen (2009), “chuoi hot” is a fruit which had high TTP content, specially the seed part Therfore, they could be considered as a ab abc bcd cd d

Seed Namdinh bcd abc ab ab a

Seed Yenbai potential source of bioactive compounds to be studied and exploited.

Effect of maturity stage to antioxidant capacity in each part of “chuoi hot”

IN EACH PART OF “CHUOI HOT”

Antioxidant capacity is an important characterictic of polyphenol compound which had been focused by researchers Antioxidant capacity was shown in feel, pulp and seeds of “chuoi hot”

Statistical analysis shows that harvest location and the interaction between maturity stage and harvest location do not significantly affect the antioxidant capacity of the peel (p = 0.599, p = 0.988) In contrast, the maturity stage has a significant effect on peel antioxidant capacity (p = 0.000) These results imply that while where the fruit is harvested and the combined effect with maturity stage have limited influence, selecting the appropriate maturity stage is crucial for maximizing the antioxidant potential of the peel.

Namdinh peel has higher antioxidant capacity (134.92 ± 19.62 µmol TE/g DW) than Yenbai peel (126.91 ± 44.79 µmol TE/g DW) Across maturity stages, peel antioxidant capacity decreases dramatically from Stage 1 to Stage 5 In Namdinh, it drops from 236.5 ± 29.0 to 68.73 ± 29.7 µmol TE/g DW, about a 3.4-fold decrease, while in Yenbai the cultivar “chuoihot” falls from 230.4 ± 87.3 to 57.24 ± 19.7 µmol TE/g DW By comparison, mango fruit/puree antioxidant capacity declines from 167.5 ± 13.4 to 123.7 ± 12.3 µmol TE/g puree during maturation (Mahattanatawee et al., 2006).

Statistics analysis result showed that maturity stage, interaction between maturity stage and harvest location significantly effected antioxidant capacity of pulp (p=0.019, p=0.000) while harvest location did not have any effect (p=0.581)

Antioxidant capacity of pulp in Yenbai (187.61 ± 48.92 àmol TE/g DW) was higher than that in Namdinh (179.51 ± 12.05 àmol TE/g DW) In Namdinh, there was a dramatic drop from 289.9 ± 17.7 àmol TE/g DW at the 1st maturity to 140.63 ± 4.34 àmol TE/g DW at the 5th maturity, a decrease of about 2.1 times; the 2nd, 3rd, 4th and 5th maturity stages recorded 163.85 ± 17.19, 157.25 ± 3.83, 145.89 ± 17.21, and 140.63 ± 4.34 àmol TE/g DW, respectively By contrast, Yenbai pulp antioxidant capacity ranged from 90.77 ± 9.09 àmol TE/g DW at the 1st maturity to 286.3 ± 61 àmol TE/g DW at the 3rd maturity, followed by a decline from the 3rd to the 5th maturity stage.

Namdinh and Yenbai on antioxidant capacity of pulp

Statistics analysis result showed that harvest location, maturity stage did not have significant effect on antioxidant capacity of seed (p=0.266, p=0.208)

Interaction between harvest location and stage signifficantly effected antioxidant capacity of seed (p=0.000) a bc bc bc bc

Pulp Namdinh c ab a abc bc

Figure 4.10 Impact of the maturity stage of the “chuoi hot” fruit harvested in Namdinh and Yenbai on antioxidant capacity of seed

As shown in Figure 4.10, the antioxidant capacity of seeds from Namdinh Province decreases markedly during maturation, while seeds harvested in Yen Bai Province increase from the first to the fifth maturity stage For Namdinh seeds, antioxidant capacity declines from 267.92 ± 11.03 µmol TE g−1 DW at the 1st maturity stage to 166.4 ± 31.1 µmol TE g−1 DW at the 5th maturity stage.

Antioxidant capacity of Yenbai seeds increased from 193.8 ± 31.3 µmol TE/g DW at the 1st stage to 250.8 ± 7.87 µmol TE/g DW at the 5th stage, illustrating a notable rise across developmental stages Similarly, papaya antioxidant capacity rose during ripening from 29.7 ± 5.4 µmol TE/g puree to 65.1 ± 15.8 µmol TE/g puree (Jimenez-Escrig et al., 2000).

Antioxidant capacity varies among different parts of chuoi hot harvested in Namdinh and Yenbai Mahattanatawee et al (2006) reported the antioxidant capacities of Florida fruits as follows: red guava 609.3 ± 31.9 µmol TE/g puree, white guava 298.6 ± 22.6 µmol TE/g puree, red dragon 134.1 ± 30.1 µmol TE/g puree, white dragon 34 ± 7.3 µmol TE/g puree, papaya 65.1 ± 15.8 µmol TE/g puree, and logan 69.6 ± 15.8 µmol TE/g puree In comparison, our results indicate that chuoi hot possesses a high antioxidant capacity relative to these fruits, suggesting its potential as a source of phenolic antioxidant compounds for food and drug technology.

Piceatannol content of seed in maturity stage

Seed Namdinh bcd bcd abcd abc ab

C Figure 4.11 Chromotography of pulp(A), peel (B), seed (C)of “chuoi hot” harvetsed in Namdinh at 1 st maturity stage

Figure 4.12 shows that the harvest location—Namdinh and Yenbai—significantly affected the piceatannol content in the seeds of the “chuoi hot” fruit, with a p-value of 0.000 By contrast, the maturity stage and the interaction between harvest location and maturity stage did not have significant effects on piceatannol content (p = 0.670 and p = 0.786, respectively).

Average picetannol content in seed of “chuoi hot” harveted in Namdinh (0.19± 0.05mg/g DW) was higher the one of seed harvested in Yenbai (0.09±

0.03mg/g DW) Piceatannol content of seed collected in Namdinh were 0.219 ± 0.1(mg/g DW), 0.176 ± 0.06 (mg/g DW), 0.177 ± 0.06(mg/g DW), 0.215 ± 0.01(mg/g DW),0.151 ± 0.04 (mg/g DW) for the 1 st , 2 nd , 3 rd , 4 th and 5 th maturity stages, respectively Piceatannol content of seed harvested in Yenbai at different maturity stage were as following: 0.096 ± 0.057 (mg/g DW), 0.076 ± 0.03(mg/g DW), 0.087 ± 0.03(mg/g DW), 0.084 ± 0.02(mg/g DW), 0.088 ± 0.02(mg/g DW)

Piceatannol is found in red wine and grapes at high concentrations A 2013 study by Lai et al reported a high piceatannol content in certain fruits, measured at 2.30 ± 0.01 mg/g dry weight Blueberries contain about 186–422 ng/g dry weight, according to Rimado et al (2004), while red grapes show piceatannol levels of 0.27–0.54 µg/g fresh weight (Guerrero et al., 2010) In addition, piceatannol has been detected in other fruits such as passion fruit and certain Asian beans.

Compared with other plants, Chuoihot exhibits a high concentration of piceatannol Its piceatannol content is 450–1000 times higher than blueberry and 350–700 times higher than red grape Consequently, seeds of Chuoihot may be considered a notably rich source of piceatannol.

“chuoi hot” became a newnatural source of piceatannol which can be used in food and drug technology a a a a a

Pi ce at an no l c on te nt (m g/ g CK )

Conclusion and recommendation

Conclusion

In this study, the detailed of physical-chemical properties and chemical composition including total polyphenol, antioxidant capacity and piceatannol content of Musa babilsiana were determined The results shown that:

In “chuoi hot”, the propotion of pulp was the highest and the one of seed was the lowest

During maturation, the ratio of peel decreased, the portion of pulp increased, while the percentage of seed did not change

The hardness of pulp fall during maturation

“Chuoihot” harvested in Namdinh and Yenbai contented two monosaccharids including glucose and fructose The sugar content increased during maturity

Harvest location and maturity stage significantly influence phenolic content and antioxidant capacity in the peel, pulp, and seed of chuoi hot In Namdinh, maturation is associated with a decrease in total phenolic content and antioxidant capacity across peel, pulp, and seed By contrast, chuoi hot harvested in Yenbai shows a different pattern, with the peel, pulp, and seed exhibiting varied changes in phenolic content and antioxidant capacity during maturation.

Piceatannol, a potent bioactive stilbene, has been detected in the seeds of chuoi hot for the first time, with high concentrations found specifically in the seed tissue The findings show that the maturity stage does not affect seed piceatannol content, while harvest location does, indicating environmental factors influence accumulation This discovery positions chuoi hot seeds as a new source of the health-promoting compound piceatannol, warranting further study, development, and potential applications in nutraceuticals and functional foods.

Recommendation

Chuoi hot has high phenolic antioxidant content, and to date only piceatannol has been identified in this fruit; further research is needed to identify the full spectrum of phenolic compounds present These findings could explain its use in Vietnamese traditional medicine and may open new opportunities for the development of this wild fruit in the future.

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General Linear Model: % seed versus province, stage

Factor Type Levels Values province fixed 2 ND, YB stage fixed 5 1, 2, 3, 4, 5

Analysis of Variance for % seed, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P province 1 400.32 400.32 400.32 9.90 0.005 stage 4 44.01 44.01 11.00 0.27 0.893 province*stage 4 179.93 179.93 44.98 1.11 0.378 Error 20 808.93 808.93 40.45

Obs % seed Fit SE Fit Residual St Resid

R denotes an observation with a large standardized residual

Grouping Information Using Tukey Method and 95.0%

ND 15 14.83 B Means that do not share a letter are significantly different

Means that do not share a letter are significantly different

Confidence province stage N Mean Grouping

General Linear Model: % pulp versus province, stage

Analysis of Variance for % pulp, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P province 1 7667.38 7667.38 7667.38 506.58 0.000 stage 4 172.24 172.24 43.06 2.84 0.051 province*stage 4 82.05 82.05 20.51 1.36 0.285

Obs % pulp Fit SE Fit Residual St Resid

General Linear Model: % peel versus province, stage

Analysis of Variance for % peel, using Adjusted SS for Tests

Obs % peel Fit SE Fit Residual St Resid

General Linear Model: Hardness versus province, stage

Analysis of Variance for Hardness, using Adjusted SS for Tests

Obs Hardness Fit SE Fit Residual St Resid

General Linear Model: Glucose versus province, stage

Analysis of Variance for Glucose, using Adjusted SS for Tests

Obs Glucose Fit SE Fit Residual St Resid

General Linear Model: Fructose versus province, stage

Factor Type Levels Values province fixed 2 ND, YB stage fixed 5 1, 2, 3, 4, 5 Analysis of Variance for Fructose, using Adjusted SS for Tests

Obs Fructose Fit SE Fit Residual St Resid

General Linear Model: PPT peel versus province, stage

Analysis of Variance for PPT peel, using Adjusted SS for Tests

Unusual Observations for PPT peel

Obs PPT peel Fit SE Fit Residual St Resid

General Linear Model: PPT pulp versus province, stage

Analysis of Variance for PPT pulp, using Adjusted SS for Tests

Unusual Observations for PPT pulp

Obs PPT pulp Fit SE Fit Residual St Resid

1 6 15.393 C Means that do not share a letter are significantly different

General Linear Model: PPT seed versus province, stage

Analysis of Variance for PPT seed, using Adjusted SS for Tests

Unusual Observations for PPT seed

Obs PPT seed Fit SE Fit Residual St Resid

ND 5 3 37.66 D Means that do not share a letter are significantly different

Results for: Worksheet 4 General Linear Model: DPPH peel versus province, stage

Analysis of Variance for DPPH peel, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P province 1 481 481 481 0.29 0.599 stage 4 109160 109160 27290 16.16 0.000 province*stage 4 521 521 130 0.08 0.988 Error 20 33769 33769 1688

Unusual Observations for DPPH peel

Obs DPPH peel Fit SE Fit Residual St Resid

General Linear Model: DPPH pulp versus province, stage

Analysis of Variance for DPPH pulp, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P province 1 493 493 493 0.32 0.581 stage 4 23583 23583 5896 3.77 0.019 province*stage 4 94413 94413 23603 15.11 0.000 Error 20 31249 31249 1562

Unusual Observations for DPPH pulp

Obs DPPH pulp Fit SE Fit Residual St Resid

General Linear Model: DPPH seed versus province, stage

Analysis of Variance for DPPH seed, using Adjusted SS for Tests

Unusual Observations for DPPH seed

Obs DPPH seed Fit SE Fit Residual St Resid

General Linear Model: Piceatannol versus province, stage

Analysis of Variance for Piceatannol, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P province 1 0.076924 0.076924 0.076924 29.52 0.000 stage 4 0.006202 0.006202 0.001550 0.59 0.670 province*stage4 0.004477 0.004477 0.001119 0.43 0.786 Error 20 0.052116 0.052116 0.002606

Obs Piceatannol Fit SE Fit Residual St Resid

General Linear Model: TTP versus Part, stage

Factor Type Levels Values Part fixed 3 peel, pulp, Seed stage fixed 5 1, 2, 3, 4, 5

Analysis of Variance for TTP, using Adjusted SS for Tests

Source DF Seq SS Adj SS Adj MS F P Part 2 30645.6 30645.6 15322.8 83.52 0.000 stage 4 1078.5 1078.5 269.6 1.47 0.220 Part*stage 8 1953.8 1953.8 244.2 1.33 0.241 Error 75 13760.2 13760.2 183.5

Obs TTP Fit SE Fit Residual St Resid

Part N Mean Grouping Seed 30 59.33 A pulp 30 21.73 B peel 30 18.80 B Means that do not share a letter are significantly different

Part stage N Mean Grouping Seed 1 6 62.36 A

Seed 2 6 61.62 A Seed 5 6 59.03 A B Seed 3 6 57.59 A B Seed 4 6 56.03 A B C peel 1 6 32.98 B C D pulp 3 6 29.63 C D pulp 2 6 27.22 D peel 2 6 21.25 D pulp 4 6 19.28 D pulp 5 6 17.12 D peel 3 6 16.06 D pulp 1 6 15.39 D peel 4 6 13.60 D peel 5 6 10.11 D

Tiêu đề	Effect of Maturity Stage and Harvest Location on Chemical Composition and Antioxidant Capacity of Extracts from Different Parts of Musa Balbisiana Colla Fruit
Tác giả	Ngo Thi Huyen Trang
Người hướng dẫn	Dr. Lai Thi Ngoc Ha
Trường học	Vietnam National University of Agriculture
Chuyên ngành	Food Science and Technology
Thể loại	Thesis
Năm xuất bản	2017
Thành phố	Hanoi

Định dạng
Số trang	80
Dung lượng	4,43 MB