Industrial Crops & Products 209 (2024) 117937

Contents lists available at ScienceDirect

Industrial Crops & Products

journal homepage: www.elsevier.com/locate/indcrop

Novel and environmentally friendly paste-bath dyeing of protein fibers with

mulberry leaf and silkworm excrement
Yijun Tang , Weibang Xia , Zhonghua Li , Qing Li *
State Key Laboratory of Resource Insects, Chongqing Engineering Research Center for Biomaterial Fiber and Modern Textile, School of Sericulture, Textile and Biomass
Science, Southwest University, Chongqing 400715, China

A R T I C L E I N F O A B S T R A C T

Keywords: Silkworm excrement is one of the most important byproducts of sericulture industry. This work investigated
Silkworm excrement chlorophyll and its derivatives from mulberry leaves and silkworm excrement through FTIR analysis. Thermal
Mulberry leaf stability of mulberry leaves and silkworm excrement were investigated by TG and DTG. Inspired by natural mud
Chlorophyll
dyeing, this work conducted eco-friendly paste-bath dyeing without extracting colorants with organic solvents.
Pheophytin
Pyropheophytin
Results show that digestive enzyme and microorganism induced formation of pyropheophytins in metabolism of
Dyeing silkworm body is similar to those conducted through acid and/or heating, and that silkworm excrement exhibits
excellent thermal stability. When used as natural dye, pyropheophytins within silkworm excrement are able to
form connection with amino acid in protein fibers, at protein iso-electric point of around pH 4.8. Silk and wool
fabrics dyed with silkworm excrement show deep green brown shade with satisfactory color fastness to wash and
light. This work provides evidence that pyropheophytin as natural dye is more thermal and light stable than
pheophytin. Functional properties such as UV protection and oxidant inhibition are also found in fabrics dyed
with silkworm excrement.

1. Introduction
Silkworm excrement (SE) is one of the most important by-products of
sericulture industry (Manjunath et al., 2020). China, India, Brazil,
Japan, Thailand, Vietnam, North Korea, and Iran are the many countries
engaged in mulberry trees planting and silk manufacturing. Currently
China and India are the world’s leading producers for silk. In China
alone, silk production accounts for 60% of global production (Man­
junath et al., 2020). During sericulture process, a large amount of SE is
produced. According to statistics, the annual production of SE in China is
as high as 5 million tons (Liu et al., 2022; Zhang et al., 2019).
Granular and dark green in color, SE has diameter of 1–5 mm and
contains many chemical components. SE is practically a product from
mulberry leaves (ML) through silkworms’ digestive system. There are
many similar chemical compositions in SE and ML. It is shown that
around 60% of the ML consumed by silkworms are excreted without
being digested, and the rest of 40% are that resulted from functionali­
zation of enzymes and microorganisms in the silkworm’s digestive sys­
tem (Li et al., 2021; Liao et al., 2020; Sohn et al., 2009). For example,
crude fat content between SE and ML are similar, whereas content of
certain long-chain fatty acids is different. Crude protein and amino acid
content in SE are significantly lower (10%) than that of ML (21%) due to
the digestive tract of silkworm on digestion and absorption of protein
and amino acids (Li et al., 2021). A number of active substances, such as
chlorophyll (and derivatives), flavonoids and carotenoids are found in
SE. Chlorophyll for instance, which accounts for 0.8–1% of dry matter
content of SE, is higher in SE than that in ML (Hu et al., 2005; Wu et al.,
2012).
Chlorophyll and carotenoids are the natural colorants presented in
SE (Uzakova et al., 1987a; Vimolmangkang et al., 2014), making the
sericulture byproduct possible to be used in textile dyeing industry. One
of the most common processes is to extract chlorophyll from SE with
organic solvents, and the extracted chlorophyll was then processed
further into sodium copper chlorophyllin (Fig. 1) by saponification and
other chemical reactions (Han et al., 2013). Sodium copper chlor­
ophyllin is a more stable colorant with better dyeing effect and higher
color fastness for dyed fabrics than chlorophyll (Farag, 2006; Mortensen
and Geppel, 2007). The process of sodium copper chlorophyllin, how­
ever, uses strong chemicals and organic solvents such as sodium hy­
droxide and acetone which can cause environmental concern. Mordant
dyeing is another technique commonly used to improve color fastness of
fabrics dyed with natural colorants extracted from SE (Vila et al., 2018).

* Corresponding author.
E-mail address: qingli@swu.edu.cn (Q. Li).

https://doi.org/10.1016/j.indcrop.2023.117937
Received 10 May 2023; Received in revised form 23 October 2023; Accepted 22 November 2023
Available online 26 December 2023
0926-6690/© 2023 Elsevier B.V. All rights reserved.
Y. Tang et al. Industrial Crops & Products 209 (2024) 117937

Mordant dyeing process, however, involves metal ions which are general
difficult to undergo treatment, therefore discharge of wastewater can
put risk to the environment.
Mud dyeing is a traditional dyeing technique from ancient China and
Japan, which uses iron in certain type of local soil to react with tannic
acid in dye. Fabric with rare dark or black shade can thus be achieved.
Inspired by mud dyeing process, this work proposes a simple and green
dyeing method of using paste bath for dyeing protein fibers of silk and
wool. Instead of extracting natural colorants from SE with organic sol­
vents, this work applies fabric dyeing directly in SE paste. The process of
making SE paste is simple and environmentally friendly with only very
small amount of water and acetic acid (for adjusting pH) needed. No
other chemicals are required. Therefore the process of dyeing and
discharge of residuals from dyeing are environmentally friendly.
Because SE is literally a product from ML, comparisons between SE and
ML are investigated in this study in terms of chemical composition (such
as chlorophyll and its derivatives), thermal stability, dyeing ability and
color fastness of dyed fabric. Functional properties of UV protection and
Fig. 2. Powdering process for mulberry leaves (ML) and silkworm excre­
antioxidant capacity are also examined for dyed fabrics in this work. ment (SE).

2. Experimental carefully taken out and put through a padding mangle twice, before
rinsed and dried in shade at room temperature.
2.1. Material preparation
2.3. FTIR analysis
SE and ML used in this work were collected from laboratory raised
mulberry silkworms and mulberry garden at Southwest University,
An ALPHA II Fourier Transform Infrared Spectrometer (Bruker, US)
Chongqing, China. SE and ML were dried in an oven and processed into
was used to measure infrared spectra of ML and SE, as well as chloro­
fine powders using an electric grinder. The powders were then sifted
phyll extracts from ML and SE. The powder samples and dry KBr powder
through a 100-mesh filter (diameter 0.15 mm) to obtain finer powders
were put into the agate mortar at a ratio of 1:100 and the two powders
with uniformity (Fig. 2).
were ground together until they were evenly mixed. The tablet was then
Undyed 100% pure silk fabric (satin) was purchased from local silk
placed in the sample chamber of Fourier transform infrared spectrom­
mill with areal weight of 96 g/m2. Undyed 100% pure wool fabric (twill)
eter and scanned at a range of 4000–500 cm− 1.
was supplied by CSIRO Australia. Fabric samples were washed with
nonionic surfactant and dried in room temperature before dyeing pro­
2.4. Thermal stability analysis
cess. Acetic acid, ABTS (2,2′-azino-bis(3-ethyl-benzothiazoline-6-sul­
fonic acid)) and potassium persulfate were of laboratory grade.
Sample powders of 5.0 ± 0.2 mg were taken and filled into a cruci­
ble, and thermal stability was evaluated using a Nicolet-10 thermogra­
2.2. Preparation of SE paste and dyeing of fabrics vimetric analyzer (Thermo Fisher, USA) at a nitrogen purge gas flow rate
of 60 ML/min with temperature raised from 40 ℃ to 800 ℃ at a rate of
Powders of SE and ML were weighed and placed in beakers. Distilled 20 K/min.
water at pH 4.8 (adjusted with acetic acid) was then slowly added in
while stirring the mixture until it reaches desired consistency (approx­ 2.5. Color measurement of dyed fabrics
imate solid-to-liquid mass ratio of 1:4). pH of the paste dye bath was
then checked again. Before conducting fabric dyeing, it was ensured that Color parameters (L*, a*, b* and K/S) of dyed fabrics were measured
the dye bath had reasonably good flow with no bubbles. by a Datacolor 650™ (Datacolor SPECTRUM, USA) at 400 nm wave­
Silk and wool fabrics with proper sizes were put into the beakers. An length. Each sample was measured 5 times, and results were averaged.
even fabric immersion in the dyeing bath was ensured. Dyeing was
conducted within beakers in a water bath thermostatic oscillator at a 2.6. Color fastness of dyed fabrics
frequency of 120 Hz. Temperature of the dye bath was raised from
ambient to 85 ℃ and maintained at 85 ℃ for 90 min, and then let cool Washing fastness of dyed fabrics was determined according to ISO
down to room temperature. Upon completing dyeing, fabrics were 105-C06:1994 standard using a DigiWASH-SS (Yuanmao Co. Ltd.,

Fig. 1. Silkworms with byproduct of silkworm excrement (SE), and chemical structures of chlorophyll and sodium copper chlorophyllin.

2
Y. Tang et al.
China), and the color parameters of the washed samples were measured
using Datacolor 650TM.
Assessment of light fastness was obtained using a Xenon accelerated
aging tester (Changken Testing Equipment Co. Ltd., China) according to
the standard test method of AATCC16–2003.
2.7. Measurement for functional properties of dyed fabrics
UV protection of dyed fabrics was measured with an YG912E Textile
Anti-ultraviolet Performance Tester (MEBON Co. Ltd., China) according
to the standard of EU 13758–2001.
ABTS radical decolorization method was used to determine antioxi­
dant activity of dyed fabrics. 7 mmol ABTS and 2.45 mmol potassium
persulfate were dissolved in 1000 ML 0.2 mol/l phosphate buffer and
allowed to stand in the dark at 25 ℃ for 15 h. Fabric samples were cut
into size of 2.54 cm2 and mixed with 50 ML of diluted ABTS solution,
and then left in a dark environment for 30 min. The absorbance of each
solution was measured at 734 nm with TU-1901 double-beam UV–vis
spectrophotometer (Beijing Purkinje General Instrument Co. Ltd,
China). Radical scavenging efficiency of ABTS was calculated using Eq.
(1) (Zhou et al., 2020).
C− S
ABTS radical scavenging activity = × 100 (1)
C
Where C refers to the absorbance of control sample, and S that of
dyed fabric.
3. Results and discussion
3.1. FTIR analysis
SE is a product from ML through silkworms’ digestive system.
Therefore, chemical components of SE are sourced from completely and
incompletely digested ML. FTIR spectra of ML and SE shown in Fig. 3a
present the characteristic spectral bands corresponding to their chemical
compositions.
It is seen from Fig. 3a that ML and SE present similar wave peaks in
FTIR spectra, indicating similarities in chemical compounds within ML
and SE. Peak at 3409 cm− 1 belongs to stretching vibration of O-H in
polyphenol and polysaccharide compounds. Two small peaks of
2925 cm− 1 and 2857 cm− 1 are due to symmetric and asymmetric
stretching vibrations of C-H in aliphatic -CH2-. Peak at 1731 cm− 1 is
from stretching vibration of C– –O in the ester group, while peak at
1636 cm− 1 is attributed to stretching vibration of C– –O in the ketone
group (Chen et al., 2022; Das et al., 2019; Jin et al., 2022). These peaks
indicate the presence of chlorophyll in SE, and is consistent with a
previous work (Kang et al., 2018).
Difference between SE and ML is seen at the peak around 1420 cm− 1
corresponding to C-N stretching vibration in the porphyrin ring. A slight
Fig. 3. a) FT-IR spectra of ML and SE, and b) FT-IR spectra of chlorophylls from ML and SE.
3
Industrial Crops & Products 209 (2024) 117937
peak shift is seen from 1406 cm− 1 (ML) to 1420 cm− 1 (SE), and the peak
end is sharper in SE spectrum than that in ML. The process of Mg
removal from chlorophyll porphyrin ring in ML could have contributed
to this change (Fig. 4). Chlorophyll is unstable when subjected to en­
zymes and microorganisms in the body of silkworm, as a result it can
derive into natural pyroderivatives of chlorophylls, such as pyro­
chlorophylls, pyropheophytins and pyropheophorbides (Matti et al.,
2009; Park et al., 2003). Pyropheophytin is green brown in color and is
generally not found in natural green leafy plants such as ML (Hu and Xu,
1989; Karcz et al., 2014). These are observed in Fig. 2.
Fig. 3b shows FTIR spectra of extracted chlorophylls from ML and SE.
Peak at 1733 cm− 1 belongs to stretching vibration of C-133 =O and C-
173 =O from ester group. Peak at 1626 cm− 1 originates from stretching
vibration of skeleton C– –C. Peak at 1446 cm− 1 corresponds to the
symmetric stretching of C-N in porphyrin ring, while peak at 1120 cm− 1
belongs to skeletal C-C vibration in porphyrin ring (Guo et al., 2021;
Kang et al., 2018). Fig. 3b shows shifts of peaks to higher positions with
C––C vibration from 1626 cm− 1 (ML) to 1645 cm− 1 (SE), and C-N vi­
bration from 1446 cm− 1 (ML) to 1458 cm− 1 (SE). C-C stretch at peak of
1120 cm− 1 is sharper for SE than that for ML. These changes occurred in
Mg taking off reaction, during which time the vibrational frequency and
intensity of porphyrin ring in chlorophyll are greatly affected (Guo et al.,
2021). Results shown in Fig. 3b further confirmed the presence of
pyropheophytin within SE.
ML is the only food for silkworms. It is known that heating and/or
acid hydrolysis can lead to Mg taking off reaction in chlorophyll
porphyrin ring. Silkworm is poikilotherm with body temperature similar
to ambient temperature (Feng, 2015). Silkworm’s gut environment is
weak alkaline (Dow, 1992). Therefore, the above observed Mg taking off
reaction can only be induced by digestive enzyme and microorganism
within the body of silkworm, which follows the sequential reaction and
proceeds according to the kinetic process shown in Fig. 4, similar to that
commonly conducted through heating and/or acid hydrolysis of chlo­
rophylls. In addition to pheophytin and pyropheophytin, a small amount
of chlorophyll from indigested ML can be presented in SE. It is noticed
that when silkworms suffer from indigestion, the excrement becomes
moisturized with a significant increase in indigested green ML debris.
These excreted chlorophylls, however, can be very unstable.
3.2. Thermal stability
TG and DTG curves for ML and SE are shown in Fig. 5, from which
thermal stability of the bio colorants can be analyzed (Table 1). TG
curves measured starting temperatures for decomposition were 225.3 ◦ C
for ML and 258.4 ◦ C for SE, during which time the slight weight loss is
mainly caused by moisture evaporation and decomposition of chloro­
phyll (Chen et al., 2021). A delay of 23 ℃ indicates a higher decom­
posing temperature for SE. Study showed that 150–400 ◦ C is the
Y. Tang et al. Industrial Crops & Products 209 (2024) 117937

Fig. 4. Kinetic process of chlorophyll Mg taking off sequential reaction.

Fig. 5. TG and DTG curves for ML and SE.

Table 1
Thermal stability of ML and SE analyzed from TG and DTG measurements.
ML
decompose starting temperature (℃) 225.3
turning temperature (℃) 255.7
temperature at maximum weight loss rate (℃) 304.2
percentage mass change (%) - 47.5
percentage mass remaining @ 796 ℃ (%) 12.4
temperature range for conversion of chlorophylls to pyropheophytins,
and pyropheophytins presented in SE do not cause mass loss in con­
version process compared to ML (Samide and Tutunaru, 2017). Simi­
larly, temperature at maximum weight loss rate calculated from TG
curves show 304.2 ℃ for ML and 322.1 ℃ for SE. Mass loss at this stage
is mainly due to the presence of abundant organic compounds in ML and
SE, such as crude protein, crude fat, crude fiber and alkaloids (Zhu et al.,
2018). There is a delay of 18 ℃ for SE when the two samples reached
their temperatures at maximum weight loss rate. These results suggest a
higher thermal stability for SE than that for ML.
In terms of mass changes, it is calculated from DTG curves a mass loss
of 47.5% for ML and 34.8% for SE. At the end of the TG curves, the
percentage mass remaining @ 796 ℃ is 12.4% for ML and 19.7% for SE.
The less mass loss and higher mass remaining for SE than that for ML
confirmed a higher stability for SE than ML.
3.3. Dyeing performance and color fastness of dyed fabrics
3.3.1. Color strength
Dyeing of wool and silk with both ML and SE was conducted in this
work, and results are shown in Fig. 6a and Table 2. It is seen that fabrics
dyed with ML showing bright green color whereas SE dyed dark green
brown. These fabric colors are similar to the natural color of their dye
SE origin (Fig. 2), indicating a successful adsorption of natural colorants
258.4 onto fabric surface in the paste dye bath.
272.7 During dye paste preparation and dyeing process, however, chloro­
322.1 phyll and its derivatives in dye could have been changed in acid con­
- 34.8
19.7
dition of pH 4.8 and in heating at 85 ℃. It is known that chlorophyll in
ML can derive into pheophytin as a result of Mg taking off reaction when
subjected to acid hydrolysis (Guo et al., 2021). Pheophytin is shown to
carry negative charges (Jin et al., 2023). When dyeing was conducted at
protein iso-electric point of around pH 4.8, the negatively charged
pheophytin can form connection with protein fiber of silk and wool
(which carry positively charged functional group of amines) through
ion-dipole interaction (Fig. 6b). This pheophytin derived from chloro­
phyll however, fails to react further into pyropheophytin through acid
hydrolysis (Hu and Xu, 1989).
Indigested chlorophyll and pheophytin in SE, however, are able to
react further into pyropheophytin when subjected to acid and heating
(Gauthier-Jaques et al., 2001), leading to an increase in pyropheophytin
in SE paste during fabric dyeing. Same as pheophytin, pyropheophytin
can bond well with amino acid such as lysine in wool through ionic
interaction shown in Fig. 6b. It is seen from Fig. 6a that fabrics dyed with
SE exhibit deeper green brown color shade than that dyed with ML due
to the presence of pyropheophytin. It is noted however, that pyro­
pheophytin can be further hydrolyzed into pyropheophorbide and
phytol etc, when it suffers from strong acid and long lasting heating
(Milenković et al., 2012).
In addition to chlorophyll derivatives, a small amount of flavonoid
and carotenoid (such as lutein and β-carotene) are known presented in

4
Y. Tang et al. Industrial Crops & Products 209 (2024) 117937

Fig. 6. Fabric dyeing performance, a) Images of color shade of fabrics dyed with ML and SE, and b) ionic interaction between pyropheophytin and lysine.

dyes. This work presented evidence that pyropheophytin used as natural

Table 2
dye is more thermal and light stable than pheophytin. Also seen in
Color characteristic value, K/S and color fastness of dyed fabrics.
Table 2 that both wash and light fastness of SE dyed wool fabric is
sample ID CIEL*a*b* values K/S color fastness relatively higher than that of silk, this could be due to the stronger
source of dye fabric L* a* b* to wash to light connection between dye and wool than that between dye and silk as
ML silk 66.52 -3.89 30.19 5.09 3 2-3 mentioned previously.
wool 63.75 -0.81 31.12 7.30 3-4 3
SE silk 57.86 2.35 32.11 8.92 4 5-6 3.4. Functional finish (UV protection and antioxidant)
wool 52.84 2.59 29.95 11.33 4-5 7-8

UV protection of dyed fabric samples are shown in Table 3 and Fig. 8.

SE (Zhu and Zhang, 2014). These natural colorants can bond well with It can be seen from Table 3 that fabrics dyed with ML and SE showing a
protein fibers in dyeing process (Fig. 7), which also contribute to the significantly reduced transmittance in the UVA and UVB regions of the
green brown color seen in SE dyed fabrics (Fig. 6a). It is seen from UV spectrum and an increase in UPF values when compared with the
Table 2 that under the same dyeing conditions, wool shows a higher undyed fabrics used as control, for both silk and wool fabrics. This could
color strength (K/S) than that of silk. This could be due to the presence of be because flavonoid and carotenoid presented in natural dye have
more functional groups, such as -OH, -COOH, and -NH2 on polypeptide strong ability to absorb UV light (Feng et al., 2007; Giokas et al., 2007;
chain and side chains of amino acid in wool that can react with dye. Verma et al., 2021). It is seen that SE dyed samples have lower UVA and
Higher degree of crystallinity of silk also contributed to this observation
(Mongkholrattanasit et al., 2009; Moore, 1953). Table 3
UV protective performance of dyed fabrics.
3.3.2. Color fastness sample ID transmittance UPF grade
Natural colorants can be unstable when they left their original nat­ fabric source of dye T(UVA) T(UVB)
ural settlement, especially chlorophyll. Fabrics dyed with ML show poor silk undyed (control) 8.20 4.41 15.25 poor
ML 2.00 1.86 44.68 good
color fastness to both wash and light as seen in Table 2. The fresh green
SE 1.76 1.65 50.77 excellent
color in ML dyed fabrics (Fig. 6a) can completely fade away after wool undyed (control) 9.29 4.32 15.68 poor
exposed to sunlight for 72 h. Table 2 shows that light fastness of SE dyed ML 2.05 1.87 45.03 good
silk and wool fabrics rated 5–6 and 7–8, which is satisfactory for natural SE 1.81 1.58 52.66 excellent

Fig. 7. Hydrogen bonding between lutein and amino acid in protein fibers.

5
Y. Tang et al.
Fig. 8. UPF and percentage oxidant inhibition values of dyed silk and wool fabrics.
UVB transmittance and higher UPF values than that of the samples dyed
with ML. This could be due to the darker shade presented in SE dyed
fabrics. It has been shown in previous studies that dark fabrics transmit
less UV light than light colored fabrics, and that the darker the shade of
any color, the higher the protection (Riva et al., 2009).
Fig. 8 reports antioxidant capacity of dyed fabric samples, where
high free radical scavenging rate represents strong antioxidant capacity.
Lutein, β-carotene, flavonoids, and chlorogenic acid presented in ML and
SE have been shown to act effectively as free radical scavengers (Andallu
et al., 2014; Burton, 1989; Park et al., 2011; Rather et al., 2020; Shahid
et al., 2017; Soobrattee et al., 2005; Sugiyama et al., 2017; Uzakova
et al., 1987b; Zhang et al., 2020). These active chemical compounds
attached to the fabrics surface during dyeing, resulting in a significant
increase in antioxidant capacity of dyed fabrics compared to the control
samples. It is seen from Fig. 8 that antioxidant properties of SE dyed
fabrics were slightly higher than that of ML dyed. For both silk and wool
fabrics investigated, silk dyed fabric showed better antioxidant proper­
ties than that of wool dyed. This could be because bio-material of silk
used as substrate has bio-function by nature.
4. Conclusion
This work demonstrated that the digestive enzyme and microor­
ganism induced formation of pyropheophytins from chlorophyll in
mulberry leaves through metabolism of silkworm body is similar to
those conducted through acid and/or heating. Silkworm excrement ex­
hibits better thermal stability than that of mulberry leaves. Inspired by
natural mud dyeing, this work successfully conducted dyeing in paste
bath without extracting colorants with organic solvents. Results show
that when used as natural dye, pyropheophytins and carotenoid (such as
lutein) within SE are able to form connections with amino acid in protein
fibers of wool and silk at protein iso-electric point of around pH 4.8.
Fabrics dyed with silkworm excrement showed deep green brown shade
with satisfactory color fastness to wash and light. This work provides
evidence that pyropheophytin as natural dye is more thermal and light
stable than pheophytin. Functional properties such as UV protection and
oxidant inhibition are also presented in silkworm excrement dyed
fabrics.
CRediT authorship contribution statement
Yijun Tang: Methodology, Investigation, Data curation, Writing –
original draft. Weibang Xia: Visualization, Software. Zhonghua Li:
Investigation. Qing Li: Conceptualization, Supervision, Writing – review
& editing.
6
Industrial Crops & Products 209 (2024) 117937
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data Availability
Data will be made available on request.
Acknowledgement
The authors acknowledge support of this study from Chongqing
Overseas Scholars Innovation Program (project ID: cx2018025).
References
Andallu, B., Shankaran, M., Ullagaddi, R., Iyer, S., 2014. In vitro free radical scavenging
and in vivo antioxidant potential of mulberry (Morus indica L.) leaves. J. Herb. Med.
4, 10–17. https://doi.org/10.1016/j.hermed.2013.10.002.
Burton, G.W., 1989. Antioxidant action of carotenoids. J. Nutr. 119, 109–111. https://
doi.org/10.1093/jn/119.1.109.
Chen, C., Huang, Y., Meng, Z., Xu, Z., Liu, P., Li, T., 2021. Multi-heteroatom doped
porous carbon derived from insect feces for capacitance-enhanced sodium-ion
storage. J. Energy Chem. 54, 482–492. https://doi.org/10.1016/j.
jechem.2020.06.025.
Chen, J.X., Huang, H., Zhang, C.L., Huang, D., Zhu, Y.Q., Chai, X.L., 2022. The
mechanism of Cd sorption by silkworm excrement organic fertilizer and its effect on
Cd accumulation in rice. J. Soils Sediment. 22, 2184–2195. https://doi.org/
10.1007/s11368-022-03208-w.
Das, D., Ghosh, R., Mandal, P., 2019. Biogenic synthesis of silver nanoparticles using S1
genotype of Morus alba leaf extract: characterization, antimicrobial and antioxidant
potential assessment. SN Appl. Sci. 1, 16. https://doi.org/10.1007/s42452-019-
0527-z.
Dow, J.A.T., 1992. pH gradients in lepidopteran midgut. J. Exp. Biol. 172, 355–375.
https://doi.org/10.1242/jeb.172.1.355.
Farag, A.A.M., 2006. Optical absorption of sodium copper chlorophyllin thin films in
UV–vis-NIR region. Spectrochim. Acta, Part A 65, 667–672. https://doi.org/
10.1016/j.saa.2005.12.026.
Feng, L., 2015. Anatomy and Physiology of Silkworm. Higher Education Press, Beijing.
Feng, X.X., Zhang, L.L., Chen, J.Y., Zhang, J.C., 2007. New insights into solar UV-
protective properties of natural dye. J. Clean. Prod. 15, 366–372. https://doi.org/
10.1016/j.jclepro.2005.11.003.
Gauthier-Jaques, A., Bortlik, K., Hau, J., Fay, L., 2001. Improved method to track
chlorophyll degradation. J. Agric. Food Chem. 49, 1117–1122. https://doi.org/
10.1021/jf000384c.
Giokas, D.L., Salvador, A., Chisvert, A., 2007. UV filters: from sunscreens to human body
and the environment. TrAC, Trends Anal. Chem. 26, 360–374. https://doi.org/
10.1016/j.trac.2007.02.012.
Guo, X., Wu, F., Cheng, T., Huang, H., 2021. Extraction of a high efficiency and long-
acting green corrosion inhibitor from silkworm excrement and its adsorption
behavior and inhibition mechanism on copper. Colloids Surf., A 631, 127679.
https://doi.org/10.1016/j.colsurfa.2021.127679.
Y. Tang et al.
Han, J., Wang, Y., Ma, J.J., Wu, Y.C., Hu, Y.T., Ni, L., Li, Y.F., 2013. Simultaneous
aqueous two-phase extraction and saponification reaction of chlorophyll from
silkworm excrement. Sep. Purif. Technol. 115, 51–56. https://doi.org/10.1016/j.
seppur.2013.04.047.
Hu, J., Zhang, Y., Li, H., Yao, T., Liu, H., Lei, H., Lu, H., 2005. Studies on the stability of
Cu, Na-chlorophyll from the Silkworm Excretion. Acta Sericol. Sin. 370–373.
https://doi.org/10.3969/j.issn.0257-4799.2005.03.030.
Hu, L., Xu, D., 1989. HPLC separation and characterization of chlorin derivatives with
intact ring V from acid degradation products of silkworm excrement crude
chlorophyll mixture. Biomed. Chromatogr. 3, 72–74. https://doi.org/10.1002/
bmc.1130030207.
Jin, Y., Tu, J., Han, X., Zhuo, J., Liu, G., Han, Y., Du, H., Wang, J., Xiao, H., 2022.
Characteristics of mulberry leaf powder enriched with γ-aminobutyric acid and its
antioxidant capacity as a potential functional food ingredient. Front. Nutr. 9 https://
doi.org/10.3389/fnut.2022.900718.
Jin, Z., Emiezi Agarry, I., Li, Y., Ding, D., Cai, T., Chen, K., 2023. In vitro bioaccessibility
evaluation of pheophytins in gelatin/polysaccharides carrier. Food Chem. 408,
135252 https://doi.org/10.1016/j.foodchem.2022.135252.
Kang, Y.R., Park, J., Jung, S.K., Chang, Y.H., 2018. Synthesis, characterization, and
functional properties of chlorophylls, pheophytins, and Zn-pheophytins. Food Chem.
245, 943–950. https://doi.org/10.1016/j.foodchem.2017.11.079.
Karcz, D., Boroń, B., Matwijczuk, A., Furso, J., Staroń, J., Ratuszna, A., Fiedor, L., 2014.
Lessons from chlorophylls: modifications of porphyrinoids towards optimized solar
energy conversion. Molecules 19, 15938–15954. https://doi.org/10.3390/
molecules191015938.
Li, M.Q., Chen, L., Dai, Y.T., Li, J.C., Li, F., Li, Q., Yu, Z.H., Chai, K.Q., Zhu, Y.Q., 2021.
Differential metabolomics and network pharmacology analysis of silkworm
biotransformation between mulberry leaves and silkworm droppings. Evid. Based
Complement. Altern. Med. 2021 https://doi.org/10.1155/2021/8819538.
Liao, Q.H., Peng, X.F., Fang, H., Turng, L.S., Huang, A., Chang, C.C., 2020. Fabrication of
poly(lactic acid)/silkworm excrement composite with enhanced crystallization,
toughness and biodegradation properties. J. Polym. Environ. 28, 295–303. https://
doi.org/10.1007/s10924-019-01595-9.
Liu, Q., Zhao, S., Zhang, Y., An, X., Wang, Q., Li, S., Lin, A., Du, Y., Wei, H., 2022.
Biochar nanozyme from silkworm excrement for scavenging vapor-phase free
radicals in cigarette smoke. ACS Appl. Bio Mater. 5, 1831–1838. https://doi.org/
10.1021/acsabm.1c01080.
Manjunath, R., Kumar, A., Arun Kumar, K., 2020. Utilisation of sericulture waste by
employing possible approaches. Contam. Agric. Sources, Impacts Manag. 385–398.
https://doi.org/10.1007/978-3-030-41552-5_19.
Matti, K.M., Singh, S.S., Savanurmath, C.J., Hinchigeri, S.B., 2009. A unique red
fluorescent protein of silkworm bearing two photochromic moieties. Photochem.
Photobiol. Sci. 8, 1364–1372. https://doi.org/10.1039/b904102h.
Milenković, S.M., Zvezdanović, J.B., Anđelković, T.D., Marković, D.Z., 2012. The
identification of chlorophyll and its derivatives in the pigment mixtures: HPLC-
chromatography, visible and mass spectroscopy studies. Adv. Technol. 1, 16–24. 〈htt
ps://www.researchgate.net/publication/258696540〉.
Mongkholrattanasit, R., Kryštůfek, J., Wiener, J., 2009. Dyeing of wool and silk by
eucalyptus leaves extract. J. Nat. Fibers 6, 319–330. https://doi.org/10.1080/
15440470903354695.
Moore, W.R., 1953. The structure and properties of natural and allied fibres. J. Soc. Dyers
Colour. 69, 149–154. https://doi.org/10.1111/j.1478-4408.1953.tb02822.x.
Mortensen, A., Geppel, A., 2007. HPLC-MS analysis of the green food colorant sodium
copper chlorophyllin. Innov. Food Sci. Emerg. Technol. 8, 419–425. https://doi.org/
10.1016/j.ifset.2007.03.018.
Park, J.H., Lee, D.Y., Yun, P., Yeon, S.W., Ko, J.H., Kim, Y.S., Baek, N.I., 2011. Flavonoids
from silkworm droppings and their promotional activities on heme oxygenase-1.
J. Asian Nat. Prod. Res. 13, 377–382. https://doi.org/10.1080/
10286020.2011.555331.
Park, Y.J., Kim, W.S., Ko, S.H., Lim, D.S., Lee, H.J., Lee, W.Y., Lee, D.W., 2003.
Separation and characterization of chlorophyll degradation products in silkworm
using HPLC-UVAPCI-MS. J. Liq. Chromatogr. Relat. Technol. 26, 3183–3197.
https://doi.org/10.1081/jlc-120025517.
Industrial Crops & Products 209 (2024) 117937
Rather, L.J., Zhou, Q., Ali, A., Haque, Q.M.R., Li, Q., 2020. Valorization of natural dyes
extracted from Mugwort leaves (Folium artemisiae argyi) for wool fabric dyeing:
optimization of extraction and dyeing processes with simultaneous coloration and
biofunctionalization. ACS Sustain. Chem. Eng. 8, 2822–2834. https://doi.org/
10.1021/acssuschemeng.9b06928.
Riva, A., Algaba, I., Pepio, M., Prieto, R., 2009. Modeling the effects of color on the UV
protection provided by cotton woven fabrics dyed with azo dyestuffs. Ind. Eng.
Chem. Res. 48, 9817–9822. https://doi.org/10.1021/ie9006694.
Samide, A., Tutunaru, B., 2017. Thermal behavior of the chlorophyll extract from a
mixture of plants and seaweed. J. Therm. Anal. Calorim. 127, 597–604. https://doi.
org/10.1007/s10973-016-5490-y.
Shahid, M., Zhou, Y.Y., Tang, R.C., Chen, G.Q., Wani, W.A., 2017. Colourful and
antioxidant silk with chlorogenic acid: process development and optimization by
central composite design. Dyes Pigm 138, 30–38. https://doi.org/10.1016/j.
dyepig.2016.11.012.
Sohn, B.H., Park, J.H., Lee, D.Y., Cho, J.G., Kim, Y.S., Jung, I.S., Kang, P.D., Baek, N.I.,
2009. Isolation and Identification of Lipids from the Silkworm (Bombyx mori)
Droppings. J. Korean Soc. Appl. Biol. Chem. 52, 336–341. https://doi.org/10.3839/
jksabc.2009.060.
Soobrattee, M.A., Neergheen, V.S., Luximon-Ramma, A., Aruoma, O.I., Bahorun, T.,
2005. Phenolics as potential antioxidant therapeutic agents: Mechanism and actions.
Mutat. Res. Fundam. Mol. Mech. Mutagen. 579, 200–213. https://doi.org/10.1016/
j.mrfmmm.2005.03.023.
Sugiyama, M., Katsube, T., Koyama, A., Itamura, H., 2017. Seasonal changes in
functional component contents in mulberry (Morus alba L.) leaves. Hortic. J. 86,
534–542. https://doi.org/10.2503/hortj.OKD-053.
Uzakova, D., Kolesnik, A., Zherebin, Y.L., Evstigneeva, R., Sarycheva, I., 1987a. Lipids of
mulberry leaves and of mulberry silkworm excreta. Chem. Nat. Compd. 23, 419–422.
Uzakova, D., Kolesnik, A., Zherebin, Y.L., Sarycheva, I., 1987b. Carotenoids of mulberry
leaves and of silkworm excreta. Chem. Nat. Compd. 124–125. https://doi.org/
10.1007/BF00602482.
Verma, M., Gahlot, N., Singh, S.S.J., Rose, N.M., 2021. UV protection and antibacterial
treatment of cellulosic fibre (cotton) using chitosan and onion skin dye. Carbohydr.
Polym. 257, 117612 https://doi.org/10.1016/j.carbpol.2020.117612.
Vila, N., Silva, M., Musialak, A., Fernandes, M., Souto, A., Ferreira, A., 2018. Silkworm
excrement used for dyeing textiles. IOP Conf. Ser. Mater. Sci. Eng. IOP Publishing,
012028.
Vimolmangkang, S., Somkhanngoen, C., Sukrong, S., 2014. Potential pharmaceutical
uses of the isolated compounds from silkworm excreta. Chiang Mai J. Sci. 41,
97–104. 〈https://www.thaiscience.info/Journals/Article/CMJS/10905208.pdf〉.
Wu, Z., Kang, L., Cong, Y., 2012. Improvement on extraction of chlorophyll and synthesis
of pheophorbide from silkworm excrement by orthogonal experiment. Chin. J. Inf.
Tradit. Chin. Med. 19, 52–54. https://doi.org/10.13748/j.cnki.issn1007-
7693.2012.08.003.
Zhang, X.Z., Wang, Z.Y., Dang, G.J., Zhang, D.M., Zhou, D.F., Fan, Y.P., Yu, M., Zhang, Q.
S., Xie, J.Y., 2019. Mesoporous biochar derived from silkworm excrement for high-
performance supercapacitors. Int. J. Electrochem. Sci. 14, 8793–8804. https://doi.
org/10.20964/2019.09.34.
Zhang, Y., Deng, A.-P., Weng, Q.-Q., Lin, W.-M., Zhan, Z.-L., Huang, L.-Q., 2020. Study
on degradation of silkworm excrement chemical components in vitro. China J. Chin.
Mater. Med. 45, 2130–2137. https://doi.org/10.19540/j.cnki.
cjcmm.20200219.202.
Zhu, L., Zhang, Y.Q., 2014. Identification and analysis of the pigment composition and
sources in the colored cocoon of the silkworm, Bombyx mori, by HPLC-DAD.
J. Insect Sci. 14, 10. https://doi.org/10.1093/jis/14.1.31.
Zhou, Q., Rather, L.J., Ali, A., Wang, W., Zhang, Y., Rizwanul Haque, Q.M., Li, Q., 2020.
Environmental friendly bioactive finishing of wool textiles using the tannin-rich
extracts of Chinese tallow (Sapium sebiferum L.) waste/fallen leaves. Dyes Pigm 176,
108230. https://doi.org/10.1016/j.dyepig.2020.108230.
Zhu, Z., Liang, F., Zhou, Z., Zeng, X., Wang, D., Dong, P., Zhao, J., Sun, S., Zhang, Y.,
Li, X., 2018. Expanded biomass-derived hard carbon with ultra-stable performance
in sodium-ion batteries. J. Mater. Chem. A 6, 1513–1522. https://doi.org/10.1039/
C7TA07951F.