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Tomato-Lotus Inspired Edible Superhydrophobic Artificial Lotus Leaf

Daheng Wang, Jinxia Huang, Zhiguang Guo

PII: S1385-8947(20)32011-8
DOI: https://doi.org/10.1016/j.cej.2020.125883
Reference: CEJ 125883

To appear in: Chemical Engineering Journal

Received Date: 23 February 2020

Revised Date: 6 June 2020
Accepted Date: 10 June 2020

Please cite this article as: D. Wang, J. Huang, Z. Guo, Tomato-Lotus Inspired Edible Superhydrophobic Artificial
Lotus Leaf, Chemical Engineering Journal (2020), doi: https://doi.org/10.1016/j.cej.2020.125883

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Tomato-Lotus Inspired Edible Superhydrophobic
Artificial Lotus Leaf
Daheng Wang,1,3 Jinxia Huang, 1* and Zhiguang Guo1,2*

1. State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese

Academy of Sciences, Lanzhou 730070, China

2. Hubei Collaborative Innovation Centre for Advanced Organic Chemical Materials and Ministry

of Education Key Laboratory for the Green Preparation and Application of Functional Materials,

Hubei University, Wuhan 430062, China

3. University of Chinese Academy of Sciences, Beijing 100049, People’s Republic of China

*. Corresponding Author. Tel.: 0086-931-4968105; Fax: 0086-931-8277088. Email address:

zguo@licp.cas.cn (Guo) and huangjx@licp.cas.cn (Huang)

Key words: Superhydrophobic; Bio-Inspired; Plant waste recycle; Functional packaging;

Plastic replacement; Recycle ability

Abstract
Edible superhydrophobic surface for food related application has been fulfilled on various substrates;

however, the independent edible superhydrophobic material has yet to be realized. Here we report the

edible artificial lotus leaf (AL) fabricated through combination of superhydrophobic beeswax coating

and tomato waste recycled cutin-pectin membrane, mimicking the antifouling ability of lotus and oxygen

barrier ability of tomato. Moreover, additional heating was introduced for stability enhancement,

increasing adhesion and water vapour blocking ability. Through the combination of Merits of both lotus

leaf and tomato skin, results indicated that AL could be presented as an independent food packaging and

form straw, cup and dissolvable bag as plastic replacement material. Furthermore, AL could serve as

recyclable functional packaging for underwater storage, oxidation blocking, selective release bags.

Additionally, AL holds similar mechanical strength to tomato peel. Therefore, edible AL can provide

new insights for future edible superhydrophobic material and food packaging designs.

1
1. Introduction
Lotus leaf stands out in the vast selection of plants as its superhydrophobicity and antifouling

ability gave researcher inspirations of artificial superhydrophobic designs.[1-3] However, lotus

leaf also has been used for cooking and food serving for centuries. Ancient people utilized the

two merits of lotus leaf: antifouling and edible. However, the recent development of

superhydrophobic surfaces have been limited to low surface energy modification using harmful

perfluorinated compounds (PFCs).[4-6] Bayer et al. especially introduced safe perfluorinated

compounds for beeswax coating enhancement; however, not edible.[7] Edible

superhydrophobic food packaging material coated inside plastic container like cup could reduce

the residue the food waste, which has been researched in depth.; nevertheless, these coating

relies substrate which is often plastic or the adhesion between the coating and substrate is

poor.[8-12] In order to strengthen the adhesion, Tan et al. added an novel adhesive elastic layer

in between coating and substrate; however, the disadvantage was that the substrates were not

edible.[13] The independent edible superhydrophobic material could serve as new food

packaging that replaces plastic, as this type of material could be consumed alongside with food

content. Moreover, the superhydrophobic modification could further provide the packaging

with function of underwater storage that eliminate secondary plastic packaging for tea and

coffee.

Independent organic and biodegradable material that mimic lotus leaf was researched:

Poulikakos et al. presented independent cellulose/wax based organic and biodegradable

superhydrophobic material; however, the edible aspect and its applications were not mentioned,

but worth exploring.[14] Edible superhydrophobic material as independent packaging could

replace the plastic substrate and provide new functions like rapid-slow release, underwater

storage and edible oxidation barrier for fruit. Edible and plant derived materials like lipid and

polysaccharide could be chosen for the formation of Edible superhydrophobic material. Edible

raw material like cutin can be found in tomato as it is polymeric lipid component of

hydrophobic plant cuticle and can serve as barrier against the permeation of gas, moisture and

2
solutes due to its low affinity to water and water vapor.[15-19] Moreover, the cutin can also be

extracted from other plants.[20] Pectin is also derived from plant. The main industrial sources

of extraction for this polysaccharide are apple pomace and citrus peels.[21, 22] Furthermore,

the beeswax, mainly lipids, can be extracted from honeycomb waste after honey harvesting.

These three all materials are all edible (GRAS, 21CFR184.1973 and 1588).[8] Moreover, cutin,

pectin and beeswax can all be recycled from plants or agricultural waste; therefore, all three

raw material can be considered as accessible and low cost.

In this article, cutin from cuticle of tomato peel waste was extracted and mixed with pectin

forming edible membrane. Beeswax form honeycomb was used for superhydrophobic surface

modification and form edible artificial lotus leaf (AL). The 55 ° C heat enhancement was

introduced that heated AL could hold better underwater stability without the compromise of

water repellency. Food packaging application of the material was also researched. The AL

could form cup, straw and bag packaging. Through selective coating of the bag packaging, the

bag could serve as underwater storage bag for both liquid (honey) or solid (coffee powder and

tea leaf) foods. The AL bags could also achieve rapid release of the food content in boiled water

or against manual breach and slow release of inner content in cold water. Furthermore, the AL

could also serve as wound patch for cut apple for later consumption.

By combing merits of both tomato skin and lotus leaf, the edible AL could achieve the functions

of lotus leaf with additional oxygen and water vapour protections. Therefore, the edible AL

could serve as new functional packaging material and replacing plastic in the field of food

transportation, preservation and consumption. Furthermore, as edible AL is a mixture of water

soluble and insoluble content, abilities such as moisture induced shape transformation, recycle

ability can also be expected. In addition, water jetting test, water vapour uptake test and

mechanical test were conducted for practicality evaluation.

2. Experimental
2.1 Tomato peel cutin extraction

3
Cutin extraction was conducted according to the method proposed by Montanari et al.[23]

Tomatoes purchased from local market were processed through water boiling for 60 s. Then

the tomato skin was manually peeled and separated from seeds and pulp. After the acquisition,

tomato peels were immersed into 3 wt. % NaOH solution with pH level of 14 for cutin

saponification and solution. Then the mixture was transferred into the autoclave for 2 h of

heating at 120 C. The produce was then filtrated for the liquid phase collection; therefore,

removal of insoluble and undesired pulp, skin and suspended solids. Then the cutin can be

acquired through precipitation by acidifying the liquid phase sample to pH level around 5 – 6,

with 6 mol/L HCl solution. Finally, the cutin was collected through centrifugation of the

suspension at 10000 rpm for 15 mins. The precipitate was then washed and centrifuged twice

in the same manner, then freeze dried for cutin recovery.

2.2 The fabrication of three-dimensional superhydrophobic membrane

2.2.1 Cutin and pectin membrane forming solution (MFS) preparation

The obtained cutin was then dispensed in distilled and deionized water to form cutin solution

at 10 % w/v concentration through ultrasonication. The high-methanol pectin was dissolved in

distilled and deionized water at room temperature to form 6 % w/v solution (pH 3.2). Two types

of MFSs were acquired: 50/50 weight ratio cutin-pectin mixture solution and neat pectin

solution. The obtained MFSs were vacuumed 30 mins for degassing.

2.2.2 Bench casting of membrane

Ten grams of 50/50 weight ratio mixed cutin-pectin and neat pectin MFSs were added into petri

dish (10 cm in diameter), then put into vacuum dryer at 45 ̊C overnight to obtain membranes:

cutin-pectin membrane (CPM) and pectin membrane (PM). After casting, all membranes were

conditioned at Relative Humidity (RH) of 50 %.

2.2.3 Preparation of beeswax-in-ethanol emulsion

Two grams of beeswax (purchased from Solarbio Science & Technology Co., LTD., China)

were added into 200 ml of ethanol (purchased from Rionlon, China.). Then the mixture was

4
heated at 62 ̊C to dissolve the wax. After total dissolve of the wax was achieved, the solution

was cooled at room temperature for the precipitation of the wax particles. After precipitation

the emulsion was then treated with 2 h ultrasonication for the wax-in-ethanol emulsion.

2.2.4 Superhydrophobic surface modification

Modification was performed through spraying. The wax-in-ethanol emulsion was loaded into

spray gun (W-71, Dongcheng, China) and sprayed onto membranes. The spray gun was set at

20 psi and kept at 20 cm distance away from membranes. The spray process terminated till the

wax density achieved 2.5 mg/cm2 on one/both sides. Afterwards the Artificial lotus leaf (AL)

can be acquired. Further surface modification, mechanical stability, was reinforced through

adhesion strengthening between superhydrophobic coating and membrane: oven heating of the

coating and membrane at 55 ˚ C for 60 s. Afterwards the heated AF was conditioned at Relative

Humidity (RH) of 50 %.

2.3 Characterisation

SEM images of the membrane surface and cross section were acquired from the emission
scanning electron microscope (FESEM, QUANTA FEG 650, FEI, USA.). FTIR spectra results
were recorded from Fourier Transform Infrared Spectroscopy (FTIR, Thermo Scientific Nicolet
iS10). The mechanical properties of the membranes (55 × 5 mm2) were characterized at 25 C
through electronic universal testing machine (EZ Test EZ-LX) at speed of 0.1% / min with
loading cell of 18 N. Thickness of the membranes were previously measured in triplicate by
digital micrometre. The water contact angle (WCA) was measured through JC2000D system
(Zhongchen Digital Equipment Co., Ltd., Shanghai, China). The sliding angle (SA) was
measured using the DSA100 contact angle meter. 5 μl water droplets was applied to all WCA
and WSA tests. WCA and SA recordings of each sample were measured three times and
averaged for result.

2.4 Water vapour uptake test

Edible membranes were cut into 5 mm × 55 mm squares were dried then conditioned at 20 ˚C

and 50 % RH in a desiccator for the measurement of water vapour uptake.[24] The weight of

5
the samples was measured with time intervals of 60 mins after the placing of saturated solution

of Mg(NO3)2 in the desiccator. The water vapour uptake (w) can be calculated as:

(𝑚𝑛 ― 𝑚1)
𝑤= %
𝑚1

The mn and m1 represents the measured sample weight of each hour and the measured sample

weight of dry sample. Recordings of each sample were measured three times and averaged for

result.

3. Results and discussion

3.1 Edible artificial lotus leaf (AL) fabrication

The edible artificial lotus leaf (AL) was prepared through two steps: two-dimensional

membrane construction and three-dimensional structure construction. The two-dimensional

construction of the membrane was conducted through mixing the cutin pectin at 50/50 ratio in

acidic environment to form membrane forming solution (MFS). Cutin in MFS was extracted

through high pressure, high temperature and alkaline conditioned dissolving, then weak acidic

conditioned precipitation.[25] Then the membrane can be acquired after vacuum drying the

MFS. In the process of membrane forming, the chain of esterified Poly α1-4-galacturonic acids,

main component of high-methoxyl Pectin, can aggregate and cross-link with each other through

hydrogen bond and hydrophobic reactions at pH condition of 2.8 to 3.6 with the presence of

soluble sugar.[26, 27] At the same time the cutin can participate in this binding process with its

ester group and long aliphatic chains (Figure 1). Moreover, the membrane formation is

simultaneously triggered by the low electrostatic repulsions and low water activity under low

pH condition.[21, 28] Furthermore, due to the occurrence of hydrophobic interactions in the

process of membrane formation, the hydrophilic -OH and -NH2 groups can be exposed at the

surface of the membrane due to evaporation of the water and provide modification sites for

three-dimensional structure construction. The acquired cutin-pectin membrane (CPM) should

6
display similar appearance as tomato cuticle and mimicking the physical properties of tomato

peel.

The three-dimensional structure construction was conducted through spray coating of beeswax

onto the membrane. Beeswax contains long chain esters like palmitate could serve as an edible

surface hydrophobicity modifier.[29] The exposed ester group of the beeswax can form

hydrogen bond with the exposed -NH2, -OH groups at the membrane surface and strengthen

the bind force between beeswax coating and membrane (Figure 1). Both the appearance and

the surface morphology of CPM could be changed after beeswax modification and mimicking

the surface wettability of the lotus leaf; therefore, called artificial lotus leaf (AL).

Figure 1: Illustration of the fabrication process of cutin-pectin membrane (CPM) and artificial lotus leaf

(AL), and its forming mechanism. ss

7
3.2 Characterization of edible membranes.

Figure 2：a) FTIR results of extracted cutin, raw pectin and CPM. b) FTIR results of raw beeswax and

coated beeswax. c) Illustration of hydrogen bond in between membrane and coating. d) Image of PM,

CPM and beeswax coated SPM and AL.

To ensure the hydrogen bond binding between coating and membrane, the FTIR spectra was

conducted on extracted cutin, raw pectin and CPM (Figure 2a). Peak at 3400-3450 cm-1 of all

three spectra can be attributed to the -NH2 and -OH stretching, indicating the abundancy of

hydrogen bond binding site on the membrane. Moreover, the band at 2920 and 2850 cm-1 of

cutin and membrane spectra can be allocated to stretching vibrations of -CH3 and CH2 groups.

The peaks of -CH3 and CH2 groups at both cutin and membrane spectra indicated the existence

of cutin in the CPM. Furthermore, the FTIR spectra indicated that the CPM acquired -CH3 and

CH2 group from cutin, as the raw pectin spectra showed no obvious peaks at 2920 and 2850

cm-1. Therefore, the hydrophobicity of the membrane is mainly resulted from the long chain
8
aliphatic groups of cutin.[30] The stretching vibration at 1735 cm-1 of ester group for both cutin

and membrane spectra concurred the cutin existence.

Furthermore, the FTIR spectra of beeswax at each stage shown in Figure 2b presented the peak

at 1750 cm-1 corresponding to ester group stretching vibration. This peak indicated the existence

of ester group on the raw beeswax and beeswax coating. The band at 2920 and 2850 cm-1 of

cutin and membrane spectra can be allocated to stretching vibrations of -CH3 and CH2 group.

Both the -CH3 and CH2 group and the ester group provided the coating with hydrophobic

chemical property.[30] Moreover, the beeswax of two stages exhibited approximately identical

spectra peaks indicated that there were no major chemical changes during the spray coating

process.

As the existences of -NH2 and -OH group on membrane and ester group on beeswax coating

was confirmed by FTIR spectra, the hydrogen bond in between membrane and coating can be

verified. The enhancement mechanism was presented in Figure 2c. Both the -NH2 and -OH

group of membrane can form hydrogen bond with the ester group of beeswax coating; therefore,

strengthen the adhesion between the coating and membrane substrate.

The optical images of PM, CPM and beeswax coated SPM and AL were displayed in Figure

2d. As can be observed, the CPM and AL possessed red colour when compared with PM and

SPM. The colour change is resulted by carotenoid: lycopene an organic colour pigment from

tomato peels.[31] Moreover, the beeswax coated SPM and AL showed less glossy when under

light irradiation. This phenomenon is due to the introduced roughness from sprayed coating.

9
Figure 3: a) Surface topography SEM image of CPM with hydrophobicity and its contact angle (CA).

b) Surface topography of beeswax coated SCPM with superhydrophobicity and its CA. c) Cross-sectional

image of tomato peel cuticle. d) Cross-sectional images of SPM and SCPM.

The superhydrophobic edible membranes were further characterized through SEM images. As

presented in Figure 3a, the SEM image showed a smooth surface topography with little defects

indicating the uniform formation of CPM surface. Moreover, the CPM exhibited contact angle

(CA) of 89.5 demonstrating the significance of hydrophobic cutin for the construction of

hydrophobic membrane. As the pectin serves as binding agent in aqueous environment, the

water droplet will be absorbed into the membrane, the PM hold CA of 0 ̊.[27] As can be seen

in Figure 3b, the surface of beeswax coated AL has reached full coverage of beeswax flakes

indicating the modification of the surface wettability. The CA of 152 indicated successful

superhydrophobic modification has been achieved on the AL. Figure 3c displayed the cross-

section image of the tomato peel with thickness of ~ 30.2 μm and its outer cuticle cover with

thickness of ~ 6.87 μm. Figure 3d presented the cross-sectional images of SPM and AL. As

illustrated, the thickness of PM and CPM are ~ 5.45 μm and ~ 7.1 μm, respectively. The

thicknesses of the membranes are similar to that of tomato peel cuticle. Moreover, portion of
10
the beeswax flakes laid on top of the membranes in vertical manner, which provided the surface

with lotus leaf alike micro scale roughness and convex-concave surface pattern and further

provide the surface with superhydrophobicity (Figure S1).[32] Furthermore, the detailed

images of the membranes cross-sections indicated smooth and uniform formation of the

membranes with in. In conclusion, the SEM images and CA results further provided evidence

of successful tomato cuticle-like membrane fabrication and superhydrophobic surface

modification; moreover, indicated the artificial lotus leaf should hold similar wettability of lotus

leaf and physical property of tomato skin.

3.3 Further enhancement through oven heating

To further enhance the adhesion between coating and membrane, the oven heating process was

introduced. As can be seen in Figure 4a: the pristine AL possessed beeswax coating with flakes

around 100 nm thickness, which provided the surface with nano scale roughness and further

indicated the mechanism of superhydrophobicity gain. Moreover, after 55 ̊C heating treatment

in Figure 4b of 55 ̊C heated coating, the edge of beeswax flakes was melted with its thickness

increased to ~ 781 nm and the flakes were linked with each other; therefore, providing the

coating with stability. However, as displayed in Figure S1a and b, the melting resulted from

introduced heat could destroy the micro-nano structure, decreasing the wettability. Furthermore,

both the vertical and horizontal beeswax flakes of the have emerged spheres in diameter of 1

to 2 μm providing the coating with additional micro structure, which could be in favor of

wettability strengthening.[33, 34] As presented in Figure S2c, the wettability of the heated

membranes for different time indicated that: at point of 60 s heating, the AL maintained its

superhydrophobicity with CA at 152 and SA at 3 ̊. After that, with heating time increased, the

CA dropped and SA raised due to the destruction of micro-nano structures; therefore, the

heating time was set at 60 s. This structure changing phenomenon is mainly due to melting of

alkanes with low molecular weight (Figure S2b).[35] However, at time of 60 s and 90 s, the

emerge of beeswax spheres have balance the wettability downgrading effect of beeswax flakes

thickening and linkage (Figure S2a). Furthermore, the overall roughness has be maintained

11
after short time of heating; therefore, the heating process could increase the coating stability

without the destruction of water repelling ability of the coating. The chemical composition of

heated beeswax coating was analysed through FTIR. Peaks of the FTIR chart indicated that the

chemical composition had no major changes when compared to the unheated beeswax (Figure

S2d).

Figure 4: a) Surface morphology of pristine AL and its beeswax flake thickness. b) Surface morphology

of heated AL, its beeswax flake thickness and beeswax sphere diameter. c) Illustration of heated surface

enhancement of heating.

12
Moreover, as presented in Figure 4c, the surface of CPM possesses the hydrogen bond forming

site: -OH group. The heating process could slightly melt the beeswax flakes of the coating and

increases the contact area between coating and membrane; therefore, increasing the adhesion

between them. Furthermore, as -OH group is hydrophilic and still exposed to vapor. The

decrease of exposed area of membrane surface could endow the AL with stability against water

vapor. Further tests were explained below.

3.4 Wettability and stability tests

To further analyse the practicality and wettability of the enhanced Al and compare its stability

against other ALs, the CA and SA of various food liquid against the AL were tested (Figure

5a). Milk, tea, honey, coke, energy drinks and yogurt were selected as testing subjects, as the

selection represented different types of liquid: water-fat emulsion, alkali based, acid based,

sticky, carbonated, salty and bacteria-based food liquids. The results in Figure 5b showed

liquids with surface tension of 48 mN/m or higher can hold at least 150 CA on AL surface.

Moreover, coffee, milk and yogurt are water-fat emulsion-based food liquids with content such

as fatty acids and emulsifiers which could have impact on their wettability on AL. However,

the AL still showed ability of anti-fouling against this type of liquid with higher SA.

Furthermore, the SA results are correlated with the CA readings, as higher CA indicated better

repelling ability. Honey as an exception holds CA = 151.5 and SA = 15 ̊, the abnormal SA

could be the result of higher density of the honey. In conclusion the heated AL possessed anti-

fouling ability against all types of tested food liquids, which indicated its application potentials

for food liquid related surfaces.

13
Figure 5：a) Images of various liquids on heated AL and their flat view contact angles. b) CA and SA

of various food liquids and their surface tension. c) Water jetting stability test of ALs. d) Under water

stability test of ALs and raw membranes.

For the AL to be applied to application field like food packaging, its stability against water

impact and its underwater stability is essential. Therefore, water jetting and underwater stability

tests were conducted. As the AL surface is mainly been subjected to food liquid or aqueous

environment. The water jetting test and experiment on under water stability were conducted for

ALs. The water jetting test was performed by setting the water flow at ~ 5 m/s (12.5 kPa

according to equation below) and 20 cm above the 45 tilted ALs. The CA and SA recordings

of each sample were measured 5 times and averaged for result.

14
 2𝑃
𝑉=
𝜌

Where V, P and ρ represent water velocity (m/s), pressure (kPa) and density (kg/m3) separately.

The water flow mimicked the water splash within the container itself, as the water impact of

liquid food packaging mainly occurs within water environment of the container. As illustrated

in Figure 5c, the pristine AL showed significate CA and SA decrease after extensive water

jetting; furthermore, the AL was destroyed by the water flow after 20 mins of jetting, due to

the protection loss of outer beeswax coating and dissolve of the membrane pectin. As illustrated

in Figure S3a, the micro-scale roughness and convex-concave surface pattern was destroyed

after water jetting. However, the CA of 60 s and 180 s heated ALs retained their primary

wettability as there were no significate impact of water jet have been inflicted to the heated

ALs. The SA of 180 s heated AL was not recorded as the water droplet do not slides on this

surface, as the micro scale roughness and convex-concave surface pattern have been destroyed

during heating. But the melted beeswax formed a uniform protective layer around the

membrane even after water jetting (Figure S3c). Meanwhile the SA of 60 s heated AL had a

slight increase. This SA increase might be caused by slight loss of unattached beeswax flakes

and suppression of the convex-concave structure; however, due to slight melting of the beeswax

flakes, the coating possessed impact resistance against water flow (Figure S3b). It can be

concluded from the results that the heating process could significantly strengthen the impact

resistance of the AL.

To further explore the protective property of the coating and the underwater stability of the

heating process might brought to ALs (The ALs in this test was coated on both sides), the

underwater stability of wettability experiment was conducted (Figure 5d). PM, CPM, AL, 60

s and 180 s heated ALs were first dipped into water and tested for CA and then set immersed

into water for 48 h. The CA recordings of each sample were measured 5 times and averaged

for result. PM was dissolved instantly after the dipping. CPM dissolved after around 30 mins

of immersion, with the influence of hydrophobic cutin content. As illustrated in Figure 5d,

15
pristine AL showed mirror phenomenon at first and survived the first 12 h immersion which

indicated the existence of protective air layer in between the interface of water-AL. However,

the pristine AL with double sides beeswax coated showed initial CA drop and then gradually

dissolved after that (Figure 5d). The main reason can be concluded is that the water vapor at

the air layer could be gradually absorbed by the exposed membrane surface (indicated in figure

4c), swelling the membrane and breaking the hydrogen bond in between coating and membrane.

However, as image showed in Figure 5d, the membrane and the beeswax coating eventually

separated, as the remain of membrane sunk at the bottom and the beeswax flakes floated at the

water surface. Its underwater separation ability could endow recycle of both beeswax and

pectin-cutin mixture. Opposite to pristine AL, the 60 s and 180 s heated ALs held its wettability

after 48 h immersion. This unique under water stability could be mainly resulted by the melted

dense beeswax protective layer (indicated in figure 4c). Thus, the heating process could

significantly improve the underwater stability of the AL and provide this material with wider

application range without destroying the antifouling ability.

3.5 Slow release property observation of pristine AL

Figure 6: The slow release process of water-soluble blue dye powder in pristine AL bag (outside wall

coated).

16
As illustrated in section 3.4 the pristine AL can survive underwater for around 12 h. Therefore,

the slow release ability of inner content for pristine AL bag could be explored. As presented in

Figure 6, the AL packaging started to release content after 13 h, achieving the ability of slow

release. Before that the AL sustained its integrity after 10 h, after that the AL bag gradually

expanded and showed breach at 11 h ours 40 mins. The expand can be indicated by the red

band on the right and white scale line under the bag. The AL bag started to disform at 11 h ours

44 mins as water breached inside packaging and shown blue. As the AL is single side coated,

the water breached can further swell the bag from inside which is soluble. Therefore, as shown

at 11 h 49 mins, the winkle on AL bag disappeared on the surface when compared to 10 h AL.

The further swell caused dissolvement of the cutin-pectin membrane; therefore, breaking the

hydrogen bond between coating and substrate. As can be seen at 13 h, the coating started to

aggregate and rise to the water-air interface; furthermore, the water-soluble inner content

started to release. At 14 h, without the protection of superhydrophobic coating, the total

dissolvement of the AL packaging began. Additionally, after gentle stir, the inner blue dye has

reached full release. The slow release ability of the AL packaging at 12 h could provide new

packaging for drugs, nutritional supplements and preservatives antiseptic.

3.6 Enzymatic browning observation and Water vapour barrier property of enhanced AL

Further properties of the enhanced AL were characterised through enzymatic browning

observation and water vapour barrier test. The browning of economical fruit like apple can

leaded to nutritional loss and colour change. This browning oxidation can make apple less

marketable, causing economical loss. Therefore, it is meaningful to eliminate this oxidation by

inhibiting oxygen contact between exposed apple tissue and air. Polyphenol oxidase (PPO) as

the enzyme catalysis the enzymatic browning when tissue of plants such as apple is damaged

from cutting, partial consumption or removal of undesired part (exposure to oxygen). In the

presence of oxygen, the PPO catalysis the phenolic compounds into quinones through oxidation,

eventually resulting in formation of brown pigments.[36, 37] Thus, the observation of

enzymatic browning could indicate the oxygen barrier ability of the AL. The enzymatic

17
browning observation was conducted through the comparison of enzymatic browning degree

of non-covered apple, polyethylene (PE) covered apple and AL covered apple (contact side

uncoated). as illustrated in the Figure 7a.

All three subjects were placed at room temperature for 24 h and the non-covered apple showed

deepest browning. Further quantitative browning observation was done through RGB colour

profile measurement. Deepest colour was chosen for the RGB colour profile measurement and

comparison (Figure 7b). As shown in the left part of Figure 7b, the colour difference for non-

covered apple on colour rim is around 18 °, which indicated the enzymatic browning.

Traditionally, the plastics like PE were used in case of browning protection. As presented in

the middle part of the Figure 7b, the colour difference for PE covered apple on colour rim is

around 7 °, which indicated mitigation of enzymatic browning. As shown in right part of Figure

7b, the AL cover illustrated better browning blocking effect than PE cover when compared

with uncovered apple with colour difference of approximately -1 ° (the negative angle is

resulted by the influence of hue, saturation and value of light condition). The reason to the

better browning blocking effect might be that the AL was coated on the air contact side and

allowed uncoated side of AL adhere to the freshly cut moist apple tissue surface firmly, while

the PE cover may wrinkle.

Therefore, the browning observation indicated that AL could efficiently block the enzymatic

browning and further indicated its oxygen barrier property. Furthermore, the AL is edible,

biocompatible, degradable and can be consumed along with the rest of the apple. Therefore, the

AL could replace the plastic and eliminating the process of disposal and waste processing of

this type of cover packaging, causing no further harm to the environment.

18
Figure 7: a) Oxygen barrier test through observation of enzymatic browning degrees of membrane

covered apple (uncovered and PE covered apples as controls). b) RGB colour profile measurement and

comparison. c) water vapour uptake test of membranes for 12 hours.

The water vapour uptake test was conducted for characterization of the water vapour barrier

property of the AL and explanation of the underwater stability results of AL (ALs were double-

sides coated). As illustrated in Figure 7b, the PM possessed higher water affinity and absorbed

higher amount of water than CPM after 12 h, when comparing the water uptake percentage of

PM with that of CPM. It can be concluded that the hydrophobic cutin is a determine factor of
19
water uptake, while the pure pectin matrix is hygroscopic. Further, the water uptake percentage

of pristine AL hold a similar increase as CPM after 12 h; however, the initial curves of the two

presented different climbing rates. The difference of the initial stage could be contributed to the

disparity of exposed soluble area between pristine AL and CPM (Figure 4c). On the contrary,

the heated ALs (for both 60 s and 180 s) presented significantly low water affinity, as the water

uptake percentages of both heated ALs remained near to 0% even after 12 h. the reason to this

phenomenon might be resulted by the dense layer of beeswax cover around the CPM. The water

vapour test concurred with the under-water stability test that the dense layer of heated beeswax

coating could greatly reduce the water vapour uptake of ALs; therefore, the heated ALs could

serve as packaging for food storage in humid or underwater environments.

3.7 The moisture induced shape transformation and food packaging applications of

enhanced ALs

As illustrated in Figure 1, the membrane can be formed through aggregation and cross-linking

of pectin and cutin with each other through hydrogen bond and hydrophobic reactions. Thus,

further moisture induced shape transformation of membranes can be achieved through scribble

and evaporation of water by re-engage the aggregation and cross-linking between membranes

(Figure 8a). Moreover, when the membrane absorbs water, the chains of cutin and pectin

unlinks and separates leading to asymmetric swelling of the membrane and causing the self-

bend of membrane (Figure 8b). Furthermore, if the membrane continues to dissolve in water,

the recycle of membrane can be achieved (Figure S4). By combining both shape transformation

and self-bend property of the membrane, a series of application can be facilitated. As illustrated

in Figure 8c, such as: straw and cup can be fabricated through simple bending and moisture

induced attaching of CPM. Further application can be achieved after superhydrophobic

modifications on desired side of the membrane. As illustrated in Movie S1, the

superhydrophobic straw could hold application potentials in food packaging industry.

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Figure 8: a) Moisture induced shape transformation of CPM film, b) Self-bend ability of CPM when

applied water spray of single-side. c) Straw made from self-bended single-side coated AL and cup. d)

packaging of assembled AL containing honey, tea leaf and coffee (from left to right) (honey packaging

with outer-side coated 60 s heated ALs dipped in water) (tea leaf and coffee packaging outer-side coated

with 180 s heated AL). e) Release of honey packaging with optional release. f) Release of tea leaf and

coffee packaging in boiled water.

Furthermore, by transforming ALs into packaging, the water associated food beverage

precursor like honey, tea and coffee powder can be encapsulated for under water storage,

transport and consumption (Figure 8d). In liquid case like honey, the AL packaging can be

designed as 60 s heated Beeswax coating on both sides; therefore, AL packaging can block the

inner content from oxygen and moisture. Upon consumption, the packaging can be easily

breached and releasing the inner contents (Figure 8e and Movie S2). For solids such as tea leaf

and coffee powder, the AL packaging can be designed as 180 s heated Beeswax coating on both

sides, providing protection from moisture and oxidation (Figure 8f, Movie S3 and S4).
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Furthermore, the AL packaging can be easily dissolved into boiled water, as the beeswax melt

at temperature around 62 C. This type of AL packaging can dissolve into water after stirring or

high temperature and release the inner content; therefore, showed promising potentials for

plastic replacing edible packaging designs.

3.8 Mechanical stability

Table 1: Mechanical aspects of the tomato peel, lotus leaf and all prepared membranes. (T: Thickness

σT: tensile strength; E: elastic modulus; εB: elongation at break)

Sample T (μm) σT (MPa) E (GPa) εB (%)

PM 12 ± 7 30.4 ± 5.8 4.1 ± 0.8 1.5 ± 0.2

CPM 17 ± 9 11.8 ± 3.2 3.5 ± 0.9 0.8 ± 0.3

Lotus leaf 61 ± 12 6.5 ± 2.1 5.2 ± 1.8 4.5 ± 0.6

Tomato peels 38 ± 8 13.1 ± 5.2 0.9 ± 0.4 2.3 ± 0.7

Attached CPM 35 ± 3* 6.6 ± 2.8 3.1 ± 0.5 1.1 ± 0.5

Recycled CPM 20 ± 7 10.8 ± 2.7 3.7 ± 0.6 0.6 ± 0.2

AL 20 ± 6 11.5 ± 3.1 3.5 ± 0.4 0.7 ± 0.1

60s heated AL 21 ± 5 11.2 ± 4.3 3.2 ± 0.6 0.7 ± 0.2

180s heated AL 21 ± 2 12.5 ± 1.8 3.4 ± 0.6 0.8 ± 0.2

*overlaid part.

The membranes were further characterized with respect to their mechanical property through

measurement and calculations of thickness (T), tensile strength (σT), elastic modulus (E) and

elongation at break (εB). As indicated in Table 1, the CPM attained similar mechanical

behaviour of tomato peel of which the average values of σT and E were ca. 13 MPa and 0.9

GPa, respectively. These values were in close agreement with former results[38, 39] and the

low εB indicated the mechanical behaviour of this type of material as elastic, brittle solids.[40,

41] The σT values of CPM and tomato peel were around two times greater than that of lotus leaf.

This could be resulted by the water content loss of lotus leaf. However, higher E and εB values

were observed, as the existence of leaf vein brought lotus leaf with higher elastic property. The

PM showed rigid mechanical resistance than CPM and tomato peel, mainly due to that the
22
pectin contributed to membrane forming, while the cutin along did not exhibit this membrane

forming ability.[39] In conclusion, the introduction of hydrophobic cutin improved the water

resistance of CPM without compromising the mechanical strength and retained it to the level

similar as tomato peel.

Mechanical results of attached CPM attained similar E and εB readings when compared to

original CPM; however, the σT value were significantly lower as the thickness roughly doubled.

Moreover, the recycled CPM resembled the values of original CPM, indicated no significant

changes of mechanical strength to the CPM after the recycle process. In addition, the ALs all

presented similar mechanical behavior as CPM indicating that, both the coating and heating

processes of AL introduced no significant mechanical changes to the CPM membrane itself.

Overall, it can be concluded that the prepared ALs resemble the mechanical strength of CPM

and tomato peel and hold higher tensile strength than lotus leaf (bending ability indicated in

Figure S5). Moreover, as successful superhydrophobic modification and stability enhancement

had been achieved and illustrated in former sections, the ALs prepared successfully mimicked

the mechanical strength of tomato peel and the antifouling ability of the lotus leaf.

4. Conclusion
In summary, the edible superhydrophobic AL was produced by superhydrophobic beeswax

coating and tomato waste recycled cutin-pectin membrane, achieving the water repellency of

lotus surface, vapor barrier property of tomato peel and some chemical composition of both

plants (edible). Experiment results indicated the ALs possessed repellence for various food

liquids and hold potentials for food and packaging applications such as straw and cup. A series

of ALs were fabricated and can be applied for various food applications. The ALs possessed

merits such as: similar mechanical strength of tomato skin, edible ability, superhydrophobicity,

oxygen and water/water vapour barrier property, shape transformation ability, underwater

storage ability, rapid-slow release ability, selective release ability and recycle ability. It can be

suited for situations such as plant waste recycling, plastic replacement, packaging with ability

to slow or subjective release of inner content. Thus, the edible AL introduced in this article
23
could broaden the view of bioinspiration derived design and provide guideline for future

functional packaging material design.

Acknowledgments:

The authors thank the National Natural Science Foundation of China (Grant No. 51905520,

51675513, 51735013).

24
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Table of Content

Surface of plants holds huge research potentials: Lotus leaf holds antifouling ability resulted from its

surface wax structure and skin of fruits like tomato possesses the outer cuticle providing the fruit with

28
protection against rotting. By combining the merits of these two plants: applying main component of

cuticle – cutin with membrane forming agent pectin and introducing superhydrophobic beeswax particles

modification, the edible artificial lotus leaf (AL) can be fabricated. Moreover, additional heating was

introduced for surface stability enhancement. The edible AL could possess application potentials in fields

of plant waste recycle, functional packaging, plastic replacement and food application with the additional

merits that lotus leaf and tomato skin lacks: assemble and recycle ability.

Highlights

1. An edible superhydrophobic AL achieved the wettability of lotus surface;

2. Experiment results indicated the ALs possessed repellence for various food liquids;

3. The 60 s heated AL hold better stability against water impact;

4. The 60 s heated AL can serve as edible seal for freshly cut apple.

Conflicts of interest

There are no conflicts to declare.

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