RESEARCH ARTICLE

www.starch-journal.com

Noodle Production for Diabetics from Modified Green

Banana Starch by Phosphate Cross-Linking
Minh-Tan Vu, Kim-An Thi Nguyen, Mai-Huong Thi Pham, Hong-Nhung Thi Le,
Ngoc-Thanh Nguyen, Thanh-Tung Nguyen,* Thu-Ha Thi Pham, Trung-Duc Nguyen,
Ngoc-Tuan Nguyen, and Phan-Hang Nguyen

1. Introduction
The development of Green banana (GB) starch in prebiotic foods to address
health problems such as diabetes. It is modified by phosphate cross-linking to Digestion of starch is a multi-enzyme
process with about 40% of starch partially
increase the resistant starch content, improve functional properties, and
digested by salivary 𝛼-amylase in the mouth
reduce digestible starch consumption. The starch is modified using a mixture and the remaining is digested by pancreatic
of sodium trimetaphosphate (STMP) and sodium tripolyphosphate (STPP) 𝛼-amylase secretion of the pancreas.[1]
with 6 to 12 wt%ratios in the presence of sodium sulfate. The results showed In the intestine, sucrase and isomaltase
an increase in phosphorus content from 0.18% to 0.45% corresponding to the (from sucrose-isomaltase), and maltase and
glucoamylase (from maltase-glucoamylase)
increase in sodium trimetaphosphate/sodium tripolyphosphate ratios.
continue to hydrolyze starch into free
Resistant starch reached 60 wt% after increasing the concentration of glucose which provides energy for cells,
phosphate agent above 10 wt% and rapidly digestible starch decreased tissues, and organs in the body. The starch
significantly. Accordingly, the physicochemical properties of starch changed undergoing the digestive process is called
significantly. Phosphate cross-linking led to disruption of the crystalline digestible starch,[1] classified into rapidly
structure of starch granules and fragmentation, reducing enthalpy and digestible starch (RDS) and slow digestible
starch (SDS) according to their speed of
increasing gelatinization temperature. The oil and water absorption and
digestion in the intestine. The portion of
freeze-thaw stability of treated starch increased. While water absorption and starch incapable of hydrolysis by enzymes
solubility index of starch decreased significantly as the result of phosphate in the small intestine and has similar prop-
cross-linking. Modified starch is used for the production of rice noodles. From erties to soluble fiber is known as resistant
10 to 30 wt% of 10P modified starch mixed, cooking fracture rate is not more starch (RS).[2,3]
In vivo, studies suggest that the presence
than 10% and resistant starch content is over 41 wt%.
of RS in the diet brings health-promoting
effects against diabetes and reduction of
heart diseases.[4,5] Indeed, it can reduce
the risk of colon cancer, elevating butyrate levels, increasing the
growth of favorable microbes, and reducing glucose and insulin
M.-T. Vu, K.-A. T. Nguyen, M.-H. T. Pham, H.-N. T. Le, N.-T. Nguyen responses.[1,5,6] RS releases energy partly in the small intestine in
Faculty of Chemical Technology the form of glucose and partly in the large intestine in the form
Hanoi University of Industry
of fermentation byproducts such as acetate, thereby creating the
298 Cau Dien, Bac Tu Liem, Hanoi 100000, Vietnam
feeling of satiety and providing energy balance for a long time
T.-T. Nguyen, T.-H. T. Pham, T.-D. Nguyen
Institute of Chemistry after consumption.[2,6]
Vietnam Academy of Science and Technology RS can be classified into naturally occurring types I and II,
18 Hoang Quoc Viet, Cau Giay, Hanoi 100000, Vietnam type III is physically modified, type IV is from chemical treatment
E-mail: [email protected] such as etherification, esterification, and phosphorylation, and
T.-T. Nguyen, P.-H. Nguyen type V is from a complex of amylose and lipids.[6] The research
Graduate University of Science and Technology
Vietnam Academy of Science and Technology on RS type IV is still limited recently, e.g., with 15 results for
18 Hoang Quoc Viet, Cau Giay, Hanoi 100000, Vietnam “resistant starch type IV” in 10 years on PubMed. To increase
N.-T. Nguyen RS-type IV, phosphorylation is the effective method to modify the
UMR 8640 – Pasteur, Department Chemistry starch granule structure leading to resist effectively the approach
École normale supérieure – PSL, 24 Rue Lhomond of digestive enzymes.[7]
Paris 75005, France
Green banana (GB) contains about 60–80 wt% (d.b.) starch[8]
and a high content of resistant starch II-type (≈50 g RS 100 g−1
The ORCID identification number(s) for the author(s) of this article natural starch).[9] It is a biosource for preventing chronic dis-
can be found under https://doi.org/10.1002/star.202400001
eases such as colon cancer, high cholesterol, diabetes, and
DOI: 10.1002/star.202400001

Starch - Stärke 2025, 77, 2400001 2400001 (1 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

obesity.[8] GB starch can be modified to increase RS content by Table 1. Degree of substitution (DS) and constituents of phosphorylated
physical (microwave irradiation,[10] heat–moisture treatment,[11] starch. Modified green banana starch 6P, 8P, 10P, and 12P was phos-
phated by 6%, 8%, 10%, and 12% w/w of STMP/STPP (99/1, w/w).
etc.), chemical (phosphorylation, hydroxypropylation,[12] etc.), or
enzyme treatments.[13] Treated starch shows potential results
Samples DS (×10−3 ) P content RS [%] RDS [%] SDS [%]
in combating obesity-related diseases through modulating gut [%] [%]
microbiota[11] and reducing starch digestion.[14] Green banana
starch granules show outstanding advantages in terms of low Control – – 26.43a 64.16e 9.15a

amylose content, more order, and denser granule structure. Mod- 6P 9.57a 0.18a 38.70b 46.04d 14.97b
ification of green banana starch offers major advantages such as 8P 13.30b 0.25b 43.64c 32.17c 19.08c
improved freeze-thaw stability, gelatinization, transparency, and 10P 19.72c 0.37c 58.29d 15.78b 25.73d
improved resistant starch content. However, a complete under- 12P 23.94d 0.45d 60.26e 11.66a 27.93e
standing of green banana starch processing is still limited. In par- Mean values (n = 3) in the same column followed by different letters (a–e) represent
ticular, there are few results on starch properties after phosphate significant differences (p < 0.05).
cross-linking that can provide an overview of chemical starch
modification.
ules and creates strong cross-linked structures.[19] This can hin-
Noodles are one of the most popular foods in the world that
der the effects of the enzyme.[10,20–22]
are derived from starch. It then spread to the world with a variety
of products such as packaged noodles and dried noodles, along
with widespread cultural exchange between countries.[15,16] Noo- 2.2. Physicochemical Properties of Phosphorylated Banana
dles are produced by mixing, kneading flour, and then cutting Starch
and molding into the fibers. That makes noodle production as a
promising solution for using prebiotics to reduce the starch con- GB starch granules showed disk-like shapes with different sizes
tent from flour. Noodles rich in resistant starch can be an essen- (from 28 to 56 μm[13] ) with smooth surfaces (Figure 1b). Af-
tial food for obese and diabetic people.[5] ter phosphorylation, the structure of modified starch was frag-
In this work, green banana starch was modified by phosphate mented (Figure 1c–f). At high STMP/STPP content, the starch
cross-linking with phosphate agents and investigated the func- fragments agglomerated (Figure 1e,f).[21] Fragmentation oc-
tional properties. The results of the study will provide an under- curs due to the involvement of phosphate agents in the lay-
standing of the influence of various concentrations of phosphate ered structure of starch granules. The cleavage of glycosidic
agents and phosphate cross-linking on the structure and func- bonds of polysaccharides and crosslinking (Figure 1a) can cause
tional properties of green banana (Musa Paradisiaca L.) starch. fragmentation.[19]
The thermal properties of modified starch samples showed a
2. Results and Discussion gradual increase in gelatinization temperature (Tp ) (Figure 1g).
Tp of control was 68.04 °C[9,12] and phosphorylated GB starch
2.1. Effects of the Percentage of STMP/STPP on Starch Properties showed higher Tp from 74.35 °C (6P) to 84.02 °C (12P) (Table 2).
The gelatinization temperature increased can relate to the de-
Modified green banana starch 6P, 8P, 10P, and 12P was phos- gree of cross-linking[23] and the tighter the starch structure.[12]
phated by 6%, 8%, 10%, and 12% w/w of STMP/STPP (99/1, Enthalpy decreased linearly from ≈18 (control) to ≈5 J g−1 for
w/w). The phosphorus content (P%) and the degree of substitu- 12P sample (Table 2). This phenomenon can be influenced by the
tion (DS) of modified GB starch increased corresponding to the fragmentation (Figure 1c–f). Indeed, the spherical starch gran-
increase of STMP/STPP percentage. P% increased linearly from ules are multilayered in structure alternating with amorphous
0.18% of 6P to 0.45% of 12P. Similarly, DS also went up from lamella and crystalline lamella.[16] Starch granules were impacted
9.57 × 10−3 for 6P to 23.94 × 10−3 for 12P. The degree of sub- by cleavage of the polysaccharide during the reaction that oc-
stitution and P content affected the RS content in starch. The in- curs simultaneously with the phosphate cross-linking process.[19]
crease of RS content from 26.43% of control to 60.26% of 12P was Consequently, the initial structure was replaced by small crys-
observed. It is related to the decrease from 46.04% to 11.66% of talline fragments (Figure 1c–f) leading to the increase in crys-
RDS. While SDS increased from 14.97% to 27.93%. The linear tallinity and transition temperatures (To : onset temperature, Tp :
increase in phosphate substitution corresponds to %P and the peak temperature, and Te : end temperature, Table 2). Cleavage
RS content increases linearly. SDS increased slightly and RDS can take place more strongly in the amorphous region.[24] As a
decreased sharply. result, the reduction of the amorphous region increased the crys-
Starch treated with 10 wt% STMP/STPP (sample P10) showed tallinity of the starch (Table 2) and reduced the enthalpy of the
that the phosphorus content in starch (0.37% of phosphorus) did structure.[21,24] That is also related to the increase in RS content
not exceed the allowable threshold in food according to the FDA and a decrease in RDS (Table 1).[25]
(Table 1).[17,18] The relationship between DS and RS can be related X-ray diffraction pattern of control and treated samples
to the higher degree of cross-linking (Figure 1a)[12] and the more showed the peaks at 2𝜃 = 15°; 17°–18°; 23°; and 26°, charac-
steric inhibition that hindered the accessibility of 𝛼-amylase into terized as type C crystalline structure (Figure 1h).[26] The cross-
the granules. Starch phosphorylation is a process in which the linking can take place mainly in the amorphous region and has
substitution of the phosphate cross-linking agent group replaces less impact on the crystallization pattern of starch granules.[27]
the hydroxy group of amyloses and creates interchain bonds. This Indeed, the crystallinity increased linearly from ≈26% to ≈51%
breaks down the original crystalline structure of the starch gran- corresponding to the increase of STMP/STPP percentage (from

Starch - Stärke 2025, 77, 2400001 2400001 (2 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

Figure 1. a) Scheme of phosphate cross-linked starch; b–f) FE-SEM micrographs of banana starch and phosphorylated starch with different percentages
of STMP/STPP: b) control (0 wt% of STMP/STPP), c) 6P (6 wt% of STMP/STPP), d) 8P (8 wt% of STMP/STPP), e) 10P (10 wt% of STMP/STPP), and f)
12P (12 wt% of STMP/STPP), magnification 250, scale bar 100 μm; g) DSC diagram; h) X-ray diffraction patterns; and i) FTIR of natural banana starch
and different phosphorylated starch samples.

0 to 12 wt%) and the decrease of enthalpy (from ≈18 to ≈5 J g−1 ) 2.3. Stability of Phosphorylated Banana Starch
(Table 2).
FTIR showed broad bands at 3272 cm−1 of O─H bonds, 𝜐 C─H Structural changes in starch by phosphorylation can change the
bonds at 2924 cm−1 , amine I at 1644 cm−1 , and amide III at 1200– basic properties of the original starch. Swelling power (SP) was
1350 cm−1 , 𝜐 C─O at 1148 cm−1 , and 𝜐C─OH at 1079 cm−1 (car- investigated in the range of 60–90 °C. The swelling of starch
bohydrate polymers) for all samples (Figure 1i). In particular, the was less different between samples (about 1–3 g g−1 ) at 60 °C
stretching of P═O bonds at 2360 cm−1 in phosphorylated starch (Figure 2a). However, the difference was more obvious at higher
on modified starch was identified (Figure 1i).[26] temperatures. It was observed at 90 °C with about 25 g g−1 for

Table 2. Gelatinization temperature (To , Tp , Te ) of natural banana starch and phosphorylated starch and crystallinity of starch samples with: Control is
green banana starch non-modification; 6P, 8P, 10P, 12P samples are phosphorylated banana starches at different concentrations of phosphate agents
(STMP/STPP, 99:1, w/w): 6 wt% (6P), 8 wt% (8P), 10 wt% (10P), and 12 wt% (12P).

Sample To , [°C] Tp , [°C] Te , [°C] ΔT = Te – To , [°C] ΔH, [J g−1 ] Crystallinity, [%]

Control 64.33 68.04 71.56 7.23 17.95 25.98

6P 68.42 74.35 78.51 10.09 10.20 33.99
8P 71.35 77.26 83.14 11.79 9.55 39.84
10P 75.83 82.78 88.46 12.63 7.79 45.38
12P 78.55 84.02 93.32 13.77 4.74 51.42

Starch - Stärke 2025, 77, 2400001 2400001 (3 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

Figure 2. a) Swelling power (SP), b) water solubility index (WSI), c) water/oil absorption capacity (WA/OA), and d) rate of water loss by freeze-thaw
cycle of control and phosphorylated GB starch 6P, 8P, 10P, and 12P.

control and a linear decrease of modified samples to 18, 15, 12, centrations of STMP/STPP showed lower water loss. That can
and 10 g g−1 for 6, 8, 10, and 12P; respectively (Figure 2a). be the effect of the interaction between phosphate groups as
The water solubility of the modified starch samples was there- the positive charges leading to reducing the dehydration of
fore also significantly reduced (Figure 2b) compared to the con- starch.[10]
trol. That is observed at any temperature between 60 and 90 °C. Four cycles were performed to determine the freeze-thaw sta-
The WSI of the modified samples decreased from 1.5 to 2.5 times, bility of the starch. The release of water molecules through the
depending on the degree of phosphorylation. freeze-thaw process was represented by the syneresis values. In
The water absorption capacity (WAC) of modified starch sam- cycle 1, syneresis values gradually decreased from ≈16% (control)
ples was higher (from 1.81 g mL−1 for 6P to 2.91 g mL−1 for 12P) to <10% (8P, 10P, and 12P) (Figure 2d). Water release increased
than control (about 1.38 g mL−1 ). Similarly, the oil absorption ca- from cycle 1 to cycle 4. Indeed, syneresis increased linearly from
pacity (OAC) of modified samples was higher than the control 16% to 50% (3.2 times) for the control, 3 times for the 8P sam-
with 1.41, 1.72, 2.04, and 2.11 g mL−1 for 6P, 8P, 10P, and 12P, ple, and 2.7 times for the 10P and 12P samples (Figure 2d). The
respectively (Figure 2c). The WAC of modified starch samples release of water molecules in sample 12P was the lowest and the
differed greatly with oil absorption capacity. Meanwhile, the con- difference syneresis values between cycles of 10P and 12P were
trol sample showed insignificant differences, e.g., WA/OA of 12P lower than others. It showed that starch treated with a high con-
difference was up to ≈1.4 times but ≈1.1 times for the control centration of phosphate reagent (10–12 wt% STMP/STPP) signif-
(Figure 2c). icantly reduced water release from the starch gel.
With a higher degree of substitution, the water penetration was High freeze-thaw stability is associated with a decrease in
more difficult.[24] Indeed, the reduction of the amorphous region amylose content and an increase in amylopectin content in
by hydrolysis in the presence of STMP/STPP (discussed in Sec- starch.[12,27] This shows that starch has functional properties ben-
tion 3.2) and the crystalline fragment formed by phosphate cross- eficial for food production such as transparency, good mechanical
linking increased the crystallinity.[28] However, the fragmentation properties, and resistance of hydrolytic degradation.[23]
increased the free spaces in starch which can increase the water Phosphate cross-linking and hydrolysis can take place simul-
and oil absorption.[9,23] taneously in a reaction that changes the original starch gran-
The rate of water loss after each freeze-thaw cycle is re- ule structure with a decrease in amorphous regions and the for-
lated to the freeze-thaw stability of the starch. Lower dehydra- mation of crystalline fragments.[18] This change also increased
tion rates indicate higher freeze-thaw stability. The decrease crystallinity and decreased enthalpy (Table 2) leading to reduced
of freeze-thaw stability is a function of phosphorylation in swelling capacity and water absorption index (Figure 2a,b).[21]
starch. It showed a linear decrease in syneresis from ≈39% The change in crystal structure to type C (Figure 1h) after the
(6P) to ≈21% (12P) (Figure 2d). Treated samples at high con- reaction increases freeze-thaw stability.[10,24]

Starch - Stärke 2025, 77, 2400001 2400001 (4 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

Table 3. Percentage of RS, RDS, and SDS in rice noodles with different Table 4. Characteristics of noodles made from rice starch-phosphorylated
percentages of phosphorylated GB starch 10P. banana starch blends.

Samples Mass ratio, w/w RS [%] RDS [%] SDS [%] Samples Optimal Dry matter water Cooking Loss rate of
cooking time absorption rate breakage rate dry matter
Rice flour 10P starch [s] [%] [%] [%]

Control 349a 135.27a 0.38a 1.41a

Control 100 0 20.63a 60.07f 20.47f
S-P10 357b 140.53b 1.84b 1.48b
10P 0 100 58.29g 15.78g 25.73g
S-P20 360c 147.23c 3.30c 1.56c
S-P10 90 10 26.14b 55.97e 17.80e
S-P30 367d 155.10d 6.31d 1.66d
S-P20 80 20 32.61c 52.75d 14.27d
S-P40 379e 162.93e 30.13e 2.02e
S-P30 70 30 40.53d 47.87c 11.97b
S-P50 395f 168.87f 49.44f 2.58f
S-P40 60 40 46.35e 44.00b 12.08c
S-P50 50 50 51.81f 37.94a 10.17a Mean values (n = 3) in the same column followed by different letters (a–f) represent
significant differences (p < 0.05).
Mean values (n = 3) in the same column followed by different letters (a–f) represent
significant differences (p < 0.05).
loss was from ≈2% (for 40 wt% of 10P) to ≈2.6% (for 50 wt% of
2.4. Properties of Rice Noodles Containing Phosphorylated 10P) (Table 4). This can realize the different properties of modi-
Banana Starch fied GB starch and rice powder.[29,30] Specifically, although mod-
ified banana starch had a high resistant starch content, it shows
2.4.1. Determination of RS, RDS, and SDS in Rice Noodles high water absorption and gelatinization temperature, low sol-
Contained Phosphorylated GB Starch ubility, and swelling index. This can reduce the consistency and
tight binding of rice starch during cooking.[23,31] Therefore, 40–50
Green banana starch phosphorylated with a 10% STMP/STPP wt% of modified starch can significantly reduce the adhesion in
(10P) showed high resistant starch (RS), phosphorus content be- rice starch and increase the mass loss by hydration when boiled
low 0.4% (approved FDA conditions), low solubility, and swelling in water.
power. Therefore, 10P starch was used as a gradient for produc- Besides, the color of rice noodles was compared visually. The
ing rice noodles. Rice flour was mixed with various concentra- color of products became darker with the increase of modified
tions of modified starch 10P calculated by the mass of modified GB starch content (Figure A1, appendix).
starch per total mass of rice flour and modified starch, i.e., 10, An increase in modified green banana starch content in rice
20, 30, 40, and 50 wt% of modified starch 10P corresponding to noodle ingredients may lead to inhomogeneity with rice flour. In-
S-P10, S-P20, S-P30, S-P40, and S-P50 (Table 3). deed, modified banana starch consisting of crystalline fragments
The percentage RS, RDS, and SDS of rice noodles substi- with dense crystallinity by phosphate cross-linking (Figure 1) can
tuted with 10–50% phosphorylated starch 10P was determined become different in functional properties (Figure 2) compared to
in Table 3. After mixing rice flour and 10P modified GB starch, rice starch leading to uneven dispersion of the two types. That
the RS content increased linearly with the 10P content, e.g., 26% caused the high loss rate of matter and the increase in the break-
of RS for S-P10 and increased to 52% for 50% of 10P in S-P50. age rate of noodles.[31,32]
In contrast, RDS and SDS decreased gradually corresponding to
the increase of 10P content, e.g., from 56% of RDS for S-P10 to 3. Conclusion
38% for S-P50 and 18% of SDS for S-P10 to 10% for S-P50.
Green banana starch was phosphate cross-linked using sodium
trimetaphosphate and sodium tripolyphosphate (ratio 99:1,
2.4.2. Evaluation of Cooked Properties of Rice Noodles Containing w/w) as phosphate agents. The process causes rupture in the
Phosphorylated Starch crystalline–amorphous multilayer structure of the starch gran-
ules and recrystallization by phosphate cross-linking of the amy-
Noodles made from rice starch containing phosphorylated starch lose chains leads to fragmentation into smaller starch granules.
required a longer cooking time (395 s) in comparison with rice The analytical results showed an increase in resistant starch con-
noodles (349 s) (Table 4) to achieve the optimal cooking time. tent due to increased local crystallization. A 10 wt% of phosphate
However, the dry matter water absorption rate increased linearly agents were the optimal conditions that provided about 60% of
after treatment from 135% for control to 170% for 50% of 10P resistant starch and 0.37% of phosphorus content in starch. The
starch. phosphate crosslinking reduced the water solubility (from ≈20%
The distribution of the ingredients in the noodles directly im- to ≈7%) and swelling power (from ≈25 to ≈10 g g−1 ), enhanced
pacted the rate of broken noodles during the cooking process. freeze-thaw stability and water/oil absorption capacity (≈1.5–2
The cooking breakage rate of modified noodles was significantly times), and increased the gelatinization temperature (from ≈68
higher up to 49.44% for S-P50 and 30.13% for S-P40 (Table 4). to ≈84 °C) of treated starch.
Therefore, the noodles with 30% 10P starch are more acceptable Preparation of noodles using <30% phosphorylated starch 10P
with a cooking breakage rate of around 6.31%. (10 wt% phosphate agents) showed the optimal properties with
A drastic change occurred when increasing the 10P modified lower dry matter water adsorption (135–155%), cooking breakage
starch content to 40 wt% in the noodle ingredients. Dry matter rate (<6%), and dry matter loss rate (≈1.5%). The noodles have

Starch - Stärke 2025, 77, 2400001 2400001 (5 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

the potential for the needs of low-glycemic noodles intended for RS pellets were dissolved in 2 mL of 2 M KOH solution with stirring
diabetics and obese people. in an ice-water bath for 20 min. The solution was neutralized with 8 mL
sodium acetate buffer (pH = 3.8) and then was added 0.1 mL of AMG.
Then, the mixture was boiled in a water bath at 50 °C for 20 min to quanti-
tatively hydrolyzed RS to glucose. Distilled water was added to the solution
4. Experimental Section up to 100 mL to dilute the solution. Then, 0.1 mL of the solution was incu-
bated with 3 mL of glucose oxidase/peroxidase reagent (GOPOD) at 50 °C
Materials: Green banana (GB) starch (Musa Paradisiaca L.) was ex- for 20 min. The RS content was calculated based on the absorbance of the
tracted from green bananas sourced from Phu Tho province, Vietnam. sample at 𝜆 = 510 nm on a PerkinElmer Lambda 35 UV–vis Spectrometer,
GB Starch extraction was followed by Whistler et al.[13] Sodium hy- Equation (4):
droxide, sodium hydrosulfite, sodium trimetaphosphate (STMP), sodium
tripolyphosphate (STPP), sodium sulfate, and hydrochloric acid were
RDS (%) = (A × F×45) ∕W (2)
purchased from Sigma-Aldrich. K-RSTAR test kit was purchased from
Megazyme, Ireland.
Phosphorylation of Banana Starch: The phosphorylation process was SDS (%) = 100 − (RDS + RS) (3)
carried out in an aqueous slurry according to Woo et al.[33] A mixture of
sodium trimetaphosphate (STMP) and sodium tripolyphosphate (STPP) RS (%) = (A × F × 90) ∕W (4)
was prepared with a mass ratio of 99:1 (w/w). Banana starch (50 g, db),
5 g of sodium sulfate (10%, db), and STMP/STPP solution with various wherein, A was the optical absorbance of the analyzed sample compared
concentrations: 6, 8, 10, 12 wt% in 70 mL of distilled water at 25 °C. The to the blank sample; F was the conversion factor from absorbance to mi-
resulting slurry was adjusted to pH 11.0 by adding 1.0 M sodium hydrox- crograms (F = 100 μg D-glucose divided by GOPOD absorbance of 100 μg
ide and stirred at 60 °C for 1 h. Then, the slurry was adjusted to pH 6.5 standard D-glucose), W was the dry weight of each analysis (mg). The
with 1.0 M hydrochloric acid and centrifuged at 5000 rpm for 10 min. The blank sample was a mixture of sodium acetate buffer (100 mM, pH = 4.5)
sedimented starch was collected and washed with distilled water seven with GOPOD reagent (3 mL). The standard sample was a mixture of stan-
times.[33] The purified product was then dried at 40 °C for 10 h. dard D-glucose solution with GOPOD reagent.
Preparation of Rice Noodles with Phosphorylated Banana Starch: The Physicochemical Properties of Phosphorylated Banana Starch: Scanning
phosphorylated banana starch was used for preparing noodles. Rice flour electron microscopy (FESEM) was used to analyze the morphology of
(250 g) and the modified GB starch were mixed with 2.5 g of salt and 85 g starch granules using a scanning electron microscope (FM-6510LV – JEOL,
of water according to the instructions from noodle-making machine sup- Japan). Fourier transform infrared spectroscopy (FTIR) of the dried starch
plier (Media noodle making machine, MIDEA MJ-KM15Q1-401, China). sample was performed on a NEXUS 670 device (USA) in the region 4000–
The noodles created were wrapped tightly in a plastic bag for 6–8 h at 400 cm−1 using the pelleting technique with KBr. Differential scanning
room temperature before washing with water and dried at 35 °C. calorimetry (DCS) was performed on a DSC 204F1 Phoenix at measur-
Determination of Phosphorus Content and Degree of Substitution (DS) in ing conditions of 25–120 °C, speed 2 °C min−1 in a nitrogen environ-
Phosphorylated Banana Starch: The phosphorus content (w/w, dry basis) ment. The granule structure was measured on an XRD diffractometer (Shi-
was determined using the standard AOAC method.[34] Sodium carbonate madzu, model XRD – 7000, EUA) with measurement conditions: CuK∞ (𝜆
(1.5 g) was dissolved in 5 mL of distilled water in a crucible. A 2.5 g sam- = 0.15406 nm), U = 35 kV, I = 35 mA, scanning angle (2𝜃) from 5° to 80°.
ple of modified starch was added and mixed thoroughly. The mixture was The crystallinity of starch was determined from XRD using origin software.
boiled to dry so that no fume was produced. It was then combusted at Determination of Swelling Power and Water Solubility Index: Swelling
550 °C for 12 h in a furnace (LTD, ECF-12/22A, Leicestershire, UK). The power (SP) and water solubility index (WSI) were determined according
product obtained was mixed in 48 mL distilled water and 2 mL of HCl to Equations (5) and (6).[27] The starch sample was mixed with 10 mL of
25% w/v. After filtering the slurry, the solution was collected and diluted water and placed in a thermostatic bath at various temperatures: 60, 70,
with 250 mL of distilled water. Finally, 10 mL of the diluted solution was 80, and 90 °C for 30 min with vortex. The suspension was then cooled to
mixed with 2 mL vanadate-molybdate reagent and stand at room temper- room temperature before centrifuged at 1600 rpm for 15 min. The super-
ature for 45 min. The absorbance of the sample at 435 nm was measured natant was collected and dried in an oven at 110 °C for 24 h to determine
with a spectrophotometer (Beckman, DU65, Buckinghamshire, UK). The the solubility. Swelling power was determined as Equation (5) via the gain
percentage of phosphorus was calculated from the standard curve with in weight of the sample.
NaH2 PO4 ⋅H2 O as a phosphorus standard. The degree of substitution
(DS) was calculated according to Paschall et al.[35] Equation (1):
SP (g∕g) = (Gw − Sw ) ∕Sw (5)

DS = 162 × P∕ (3100 − 102 × P) (1) ( )

WSI (%) = Swg ∕Sw × 100 (6)
where, P was the % phosphorus content in the starch samples.
Determination of Rapidly Digestible Starch (RDS), Slowly Digestible Starch wherein, Swg was the mass of dissolved portion of the sample (g), Gw was
(SDS), and Resistant Starch (RS): The content of RDS, SDS, and RS was the mass of the sediment paste (g), and Sw was the mass of the initial dry
determined according to AOAC Method 2002.02. A 0.1 g starch sample sample (g, db).
was incubated with a mixture of pancreatic 𝛼-amylase and amyloglucosi- Water and Oil Absorption Capacity: A 1 g (db) of starch was mixed with
dase (PAA/AMG, 4 mL) at 37 °C with stirring for 20 min for RDS and 16 h 25 mL distilled water (for water absorption) or vegetable oil (for oil ab-
for RS. Then, 4 mL ethanol was added to the solution to terminate the re- sorption) for 1 h at room temperature. The slurry was then centrifuged at
action, the supernatant was collected by decantation and RS was collected 1500 rpm for 10 min. The supernatant was collected and weighed. Water
as the pellets on centrifugation. absorption was indicated by the difference in the weight of samples before
The supernatant containing RDS and SDS was neutralized with sodium and after absorption.
acetate buffer (100 mM, pH = 4.5) before being diluted in 100 mL of dis- Freeze-Thaw Stability: The freeze-thaw stability was carried out accord-
tilled water. After that 0.1 mL of the extracted materials were incubated ing to Huang et al.[36] Aqueous suspensions of starch samples (6%, w/v)
with 0.01 mL of AMG solution (300 U mL−1 ) at 50 °C for 20 min and were heated for 30 min to achieve full gelatinization and cooled to room
then incubated at 50 °C with 3 mL of GOPOD reagent solution for 20 min. temperature. Gel samples (10 mL) were extracted. They were frozen at
The absorbance of the sample was measured at 𝜆 = 510 nm on a Perkin- −20 °C for 24 h and then thawed at room temperature for 2 h. The samples
Elmer Lambda 35 UV–vis Spectrometer. The content of RDS starch and were centrifuged at 3000 rpm for 15 min, the supernatant was discarded,
SDS starch was calculated according to Equations (2) and (3), respectively. and the weight of the precipitate was measured. Syneresis was calculated

Starch - Stärke 2025, 77, 2400001 2400001 (6 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

according to Equation (7) Acknowledgements

Syneresis (%) = (m1 ∕m) × 100 (7) The authors would like to thank the funding from the Ministry of Science
and Technology of Vietnam (Grant No. ÐTÐL.CN-115/21).
wherein, m was the sample mass (g); m1 was the supernatant mass (g).
Cooking Properties of Noodles: The total weight (M2 , g) of 20 sticks of
noodles was measured. The optimal cooking time was determined by boil- Conflict of Interest
ing 20 noodle sticks (20 cm in length) in 500 mL of distilled water until the
central opaque core in the noodles vanished. This assessment was made The authors declare no conflict of interest.
by gently squeezing the noodles between two transparent glass slides. The
dry matter water absorption rate (DMWA) of noodles was determined ac-
cording to Mu et al.,[37] Equation (8). The noodles were cooled in distilled Data Availability Statement
water and placed on filter papers to absorb surface water. The overall mass
The data that support the findings of this study are available from the cor-
of the cooked noodles was measured (M1 , g).
responding author upon reasonable request.
The dry matter loss rate (DM loss) was identified in Equation (9).[28,23]
The obtained cooking solution was measured for the dry matter loss rate.
It was transferred into a weighing dish and dried in an oven at 105 °C until
a consistent weight (M3 ). Keywords
noodle, phosphate cross-linking, resistant starch, solubility, unripe banana
M1 − M2 (1 − W) starch
DMWA (%) = (8)
M2 (1 − W)
Received: January 4, 2024
M3 Revised: April 28, 2024
DM loss (%) = (9)
M1 (1 − W) Published online: July 10, 2024

wherein, M1 was the mass of cooked noodles (g); M2 was the mass of
noodles (g); M3 was the mass of dry matter in the cooking solution (g); W
was the moisture content of noodles (%). [1] A. P. Nugent, Nutr. Bull. 2005, 30, 27.
The noodle breakage rate was determined.[28] A total of 20 sticks of
[2] W. S. Ratnayake, D. S. Jackson, J. Food Sci. 2008, 73, 356.
noodles (20 cm in length) were boiled in 500 mL of water until reaching
[3] H. Englyst, H. S. Wiggins, J. H. Cummings, Analyst 1982, 107, 307.
the optimal cooking time. The percentage of broken noodles indicates the
[4] L. Khalili, A. Amini, Resistant Starch in Food Industry (Eds: K. Ramawat,
rate of noodle breakage.
Statistical Analysis: All experiments were performed in triplicate. J. M. Mérillon), Springer, Cham 2015.
Means and least significant differences (SD, p < 0.05) were calculated by [5] E. Fuentes-Zaragoza, M. J. Riquelme-Navarrete, E. Sánchez-Zapata,
analysis of variance (ANOVA), Bonferroni Test, and Excel. J. A. Pérez-Álvarez, Food Res. Int. 2010, 43, 931.
[6] M. G. Sajilata, R. S. Singhal, P. R. Kulkarni, Compr. Rev. Food Sci. Food
Saf. 2006, 5, 1.
Appendix [7] M. L. Stewart, M. L. Wilcox, M. Bell, M. A. Buggia, K. C. Maki, Nutri-
ents 2018, 10, 129.
[8] E. De Souza Viana, A. Dos Santos De Souza, R. C. Reis, V. J. Dos
Santos De Oliveira, Semin. Agrar. 2018, 39, 2399.
[9] T. B. Tribess, J. P. Hernández-Uribe, M. G. C. Méndez-Montealvo, E.
W. Menezes, L. A. Bello-Perez, C. C. Tadini, Lwt 2009, 42, 1022.
[10] A. Surendra Babu, G. N. Mohan Naik, J. James, A. Bin Aboobacker,
A. Eldhose, R. Jagan Mohan, J. Food Meas. Charact. 2018, 12, 2209.
[11] T. Y. Wu, S. J. Tsai, N. N. Sun, F. J. Dai, P. H. Yu, Y. C. Chen, C. F. Chau,
Int. J. Biol. Macromol. 2020, 160, 915.
[12] K. N. Waliszewski, M. A. Aparicio, L. A. Bello, J. A. Monroy, Carbohydr.
Polym. 2003, 52, 237.
[13] P. Zhang, R. L. Whistler, J. N. Bemiller, B. R. Hamaker, Carbohydr.
Polym. 2005, 59, 443.
[14] C. P. Rosado, V. H. C. Rosa, B. C. Martins, A. C. Soares, I. B. Santos,
E. B. Monteiro, N. Moura-Nunes, C. A. da Costa, A. R. P. da Mulder,
J. B. Daleprane, Int. J. Biol. Macromol. 2020, 145, 1066.
[15] N. Zhang, G. Ma, J. Ethn. Foods 2016, 3, 209.
[16] C. Li, Y. You, D. Chen, Z. Gu, Y. Zhang, T. P. Holler, X. Ban, Y. Hong,
L. Cheng, Z. Li, Trends Food Sci. Technol. 2021, 107, 389.
[17] S. Lim, P. A. Seib, Cereal Chem. 1993, 70, 137.
[18] S. R. Falsafi, Y. Maghsoudlou, M. Aalami, S. M. Jafari, M. Raeisi, Ul-
trason. Sonochem. 2019, 53, 110.
[19] V. Puchart, Biotechnol. Adv. 2015, 33, 261.
[20] Y. Xie, B. Zhang, M. N. Li, H. Q. Chen, Food Chem. 2019, 289, 187.
[21] R. Carmona-Garcia, M. M. Sanchez-Rivera, G. Méndez-Montealvo, B.
Figure A1. Noodles contained phosphorylated starch at different ingredi- Garza-Montoya, L. A. Bello-Pérez, Carbohydr. Polym. 2009, 76, 117.
ent ratios: rice flour was mixed with 10%, 20%, 30%, and 40% of modified [22] Y. Sang, P. A. Seib, A. I. Herrera, O. Prakash, Y. C. Shi, Food Chem.
starch 10P, corresponding to S-P10, S-P20, S-P30, and S-P40. 2010, 118, 323.

Starch - Stärke 2025, 77, 2400001 2400001 (7 of 8) © 2024 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

[23] D. Lin, W. Zhou, Z. Yang, Y. Zhong, B. Xing, Z. Wu, H. Chen, D. Wu, [32] Y. Wandee, D. Uttapap, S. Puncha-Arnon, C. Puttanlek, V.
Q. Zhang, W. Qin, S. Li, Int. J. Biol. Macromol. 2019, 128, 948. Rungsardthong, N. Wetprasit, Food Chem. 2015, 179, 85.
[24] M. Shi, F. Gu, J. Wu, S. Yu, Q. Gao, Starch/Staerke 2013, 65, 947. [33] K. S. Woo, P. A. Seib, Cereal Chem. 2002, 79, 819.
[25] H. Dong, T. Vasanthan, Food Hydrocoll. 2020, 101, 137. [34] R. Wongsagonsup, S. Shobsngob, B. Oonkhanond, S. Varavinit,
[26] T. J. Gutiérrez, Carbohydr. Polym. 2018, 202, 265. Starch/Staerke. 2005, 57, 32.
[27] R. Hoover, F. Sosulski, Starch - Stärke 1986, 38, 149. [35] M. Z. Sitohy, S. M. Labib, S. S. El-Saadany, M. F. Ramadan,
[28] T. Beta, H. Corke, Cereal Chem. 2001, 78, 417. Starch/Staerke. 2000, 52, 095.
[29] L. L. Yeo, P. A. Seib, Cereal Chem. 2009, 86, 210. [36] J. Huang, H. A. Schols, J. J. G. van Soest, Z. Jin, E. Sulmann, A. G. J.
[30] S. H. Koo, K. Y. Lee, H. G. Lee, Food Hydrocoll. 2010, 24, 619. Voragen, Food Chem. 2007, 101, 1338.
[31] K. Muhammad, F. Kusnandar, D. M. Hashim, R. A. Rahman, Int. J. [37] J. Mu, Y. Qi, K. Gong, Z. Chen, M. A. Brennan, Q. Ma, J. Wang, Y. Gen,
Food Sci. Technol. 1999, 34, 275. W. Lv, C. S. Brennan, LWT. 2022, 165, 113686.

Starch - Stärke 2025, 77, 2400001 2400001 (8 of 8) © 2024 Wiley-VCH GmbH