Bioresource Technology Reports 9 (2020) 100374

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Bioresource Technology Reports

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Lignin nanoparticles: Eco-friendly and versatile tool for new era T

Prakram Singh Chauhan
College of Basic Sciences and Humanities, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar 263145, Uttarakhand, India

ARTICLE INFO ABSTRACT

Keywords: Lignin is the second most abundant biopolymer after cellulose and is one of the most intriguing natural materials
Lignin for utilization across a wide range of applications. It is generated in large quantities as by-products in pulping
Lignin nanoparticle synthesis industries and biorefineries through various processes. Conversion of lignin into nanoparticles exhibit unique
Antibacterial properties because of this, recently it gained significant interest among researchers. Therefore this review is
UV absorbent
providing the proper updated information on the state of the art of different chemical, physical, mechanical
techniques which are involved in the synthesis of variety of lignin nanoparticles and their potential applications
in different fields such as antibacterial, UV adsorbent, antioxidant, hybrid nanocomposites, drug delivery ve-
hicles, bioremediation and carbon precursor etc. have been discussed.

1. Introduction
Lignin is the second most abundant biopolymer on earth, after
cellulose. According to Sameni (2015) and Schutyser et al. (2018),
biorefineries and pulping industries produce lignin as a by-product
through various processes such as kraft, organosolv, soda, enzymatic
hydrolysis and steam explosion. A target of 79 billion litres of second-
generation biofuels needs to be produced within the year 2022, as per
the U.S. Energy Security and Independence Act regulated in 2007. As
per the assumption, 355 L of bioethanol can be produced from one ton
of dry biomass and total of 223 million tons of dry biomass is processed
every year thus generating 62 million tons of lignin (Wang et al.
2019a).
Most of the lignin produced nowadays is used as boiler fuel whereas
small portion is used for the production of value-added products
(Agarwal et al. 2018). The type of plant material as well as its pro-
cessing methods decides the physico-chemical properties of lignin.
Lignin is inexpensive and possesses number of excellent features for
instance, high thermal stability, antioxidant activity, high amount of
carbon and favourable stiffness (Kai et al. 2016). However, different
types of lignin show distinct variants in terms of functional groups,
elemental composition and molecular weight (Upton and Kasko, 2016).
These advantages increased the interest among researcher for the
conversion of lignin into value-added products to be used in different
applications (Kai et al. 2016).
The lignin produced from kraft process is shown to possess particles
in the size range of 10 μm to > 100 μm however it harms the me-
chanical properties of the blends (Melro et al. 2018). Lignin Nano-
particles (LNPs) is one of the best strategies to improve the blending
properties of lignin since they possess novel characteristics such as high
surface area and improved properties compared to original material.
Furthermore they can be easily surface modify due to the availability of
huge number of functional groups like thiols, aliphatic hydroxyl and
phenolic which can be chemical modified thus enhancing their appli-
cation potential (Zhao et al. 2016). Thus, the researchers recently
shifted their interest towards the preparation of LNPs and exploring
their potential applications (Beisl et al. 2017; Figueiredo et al. 2018).
It is expected that LNPs will play a vital role in promoting lignin
valorization, similar to synthetic polymer nanoparticles contribute in
the polymer industry (Zhao et al. 2016; Ma et al. 2018). Various re-
search articles published so far emphasized the lignin conversion into
functional and useful products (Kai et al. 2016; Zhao et al. 2016;
Fortunati et al. 2016; Upton and Kasko, 2016; Roopan, 2017; Melro
et al. 2018; Agarwal et al. 2018; Ma et al. 2018; Figueiredo et al. 2018;
Wang et al. 2019a). However, none of the comprehensive review is
available which specifically focus on lignin nanoparticle synthesis and
their applications together. Therefore, In the present review, I sum-
marized all the information in two section, first section deals with
different preparation methods of LNPs synthesis and second section
deals with their potential applications in various field. The article is
intended to gain wide range of attention from the targeted audience
towards leveraging the LNPs for value-added applications.
2. Different methods of lignin nanoparticles synthesis
Recently various kinds of LNPs are being synthesized from different
resources using a combination of chemical and physical methods.
Different approaches such as polymerization, acid precipitation, solvent

E-mail address: drprakramsingh@gmail.com.

https://doi.org/10.1016/j.biteb.2019.100374
Received 4 November 2019; Received in revised form 23 December 2019; Accepted 24 December 2019
Available online 29 December 2019
2589-014X/ © 2019 Elsevier Ltd. All rights reserved.
 Table 1
Summary of different methods used in the preparation of lignin nanoparticles and their potential applications.
P.S. Chauhan

S. No. Abbrevation Morphology Raw material Diameter Shape Preparation method Brief description of method Application Reference

1 HX-40-DHP-G- Nanoparticle HX with ferulate 12.4–36.8 nm Complex patterns Polymerization HX dissolved in buffer; coniferyl/sinapyl NR Barakat et al.
HX-40-DHP-GS Nanoparticle 2.2–35 nm Complex patterns alcohol dissolve in dioxane; mixed in (2007) and Barakat
HX-90-DHP-G Nanoparticle HX without 21.8–39.8 nm Spherical presence of peroxidase. et al. (2008)
HX-90-DHP-GS Nanoparticle ferulate 16.8–47.2 nm Spherical
2 LNP Nanoparticle IAT 100–1200 nm Lignin clusters Acid precipitation IAT dissolved in EG and then HCl added. Encapsulation of hydrophobic Frangville et al.
LNP Nanoparticle IAT 50–150 nm Lignin clusters Acid precipitation IAT dissolved in high pH solution and then compounds for drug delivery. (2012)
rapidly HNO3 added.
3 CLNP Nanoparticle Protobind 2400 25–150 nm Porous Freeze-drying and Lignin dissolved in KOH solution followed NR Gonugunta et al.,
lignin microstructure stabilization by freeze drying, thermal stabilization and 2012
carbonized in a furnace.
4 NR Nanoparticle Organosolv lignin 140–147 nm Sphere Anti-solvent Organosolv lignin dissolved in acetone, As a antioxidant and free Lu et al., 2012
precipitation carried away by super critical CO2. radical scavenger.
5 LRPFs Nanoparticle- Calcium softwood 1600 nm NR NR NR As a reinforcing agent or filler Del Saz-Orozco
reinforced foams lignosulfonate in phenolic composites. et al., 2012
6 LNP Nanoparticle Protobind 2400 154–762 nm Lignin cluster Acid precipitation Lignin dissolved in EG and then HCl was NR Gupta et al., 2014
lignin added.
7 LMC Nanocapsules Kraft lignin 300–1400 nm Sphere Interfacial crosslinking Lignin cross linking at the interface of oil Encapsulation of hydrophobic Tortora et al., 2014
droplets using high-intensity ultrasonic compounds
technology.
8 Lignin-g- Nanoparticle Alkali lignin 20–50 μm Spherical Polymerization PDEAEMA-grafted lignin nanoparticles Surfactant for Pickering Qian et al., 2014a
DEAEMA prepared via ATPR. emulsions
9 NR Colloidal spheres Alkali lignin 104–110 nm Spheres Solvent exchange Acetylated lignin was dissolved in THF and Drug delivery systems or Qian et al. 2014b
water was gradually added to the solution. encapsulation of pesticides
11 NLP Nanoparticle Kraft lignin 10–30 nm Irregular shape Mechanical shearing Lignin (kraft) dissolved in water, treated NR Nair et al. 2014
using a high shear homogenizer for 4 h.

2
12 LNP Nanoparticle Sarkanda grass 0.081–0.27 nm NR Acid precipitation Lignin dissolved in water followed by NR Gilca et al. 2014
lignin heating and increase the pH by NaOH;
After adding formaldehyde solution pH
was decrease by HCl.
13 EbNPs Nanoparticle Indulin alkali 30–175 nm Non-spherical Acid precipitation IAT dissolved in EG and precipitated by Antibacterial effects; drug Richter et al. 2015
lignin HNO3. delivery vehicles and as a
absorbents for heavy metal
14 LRM Micelles Alkali lignin 100–120 nm Spheres Self-assembly Lignin dissolved in dioxane, aggregation Lignin/high-density Qian et al. 2015a
induced by adding cyclohexane. polyethylene blends
17 bio-PTT/LNP/ Nanoparticle Lignin protobind- 5.79–117 nm Cluster-forming Acid precipitation Lignin (protobind-1000) dissolved in EG, Hybrid nanocomposites Gupta et al. 2015
VGCF 1000 aggregate precipitated by HCl
18 LNP Nanoparticle Pristine lignin, 32.5–65.1 nm Spherical Acid precipitation Lignin dissolved in EG and then HCl was As a nanofiller in bio-based Yang et al. 2015a,
added. matrices, food packaging 2015b, 2015c
applications.
19 LNP Nanoparticles Kraft lignin 150–15,200 nm Spherical Microemulsion; Lignin solution added to the octane Surfactant for Pickering Nypelo et al. 2015
Interfacial crosslinking containing a mixture of Span 80, Tween 80 emulsions; Loading of silver
and 1-pentanol, and crosslinking by adding nanoparticles
epichlorohydrin.
20 ALNP, DLNP Nanoparticle Hardwood dioxane 44–164, 53–107 nm Spherical Solvent exchange; Lignin (dioxane, alkali) dissolved in As an antioxidants and UV Yearla and
lignin, alkali lignin Nanoprecipitation acetone/water mixture (9: 1, v/v), protectants Padmasree 2016
precipitated in water.
21 LNP Nanoparticle Acetic acid lignin 10–900 nm Spherical and Solvent exchange; Acetylated lignin was dissolved in THF and NR Kai et al. 2015
aggregate like Nanoprecipitation water was gradually added to the solution.
cluster
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Bioresource Technology Reports 9 (2020) 100374
 Table 1 (continued)

S. No. Abbrevation Morphology Raw material Diameter Shape Preparation method Brief description of method Application Reference
P.S. Chauhan

22 LNP Nanoparticle Wheat straw and 10–10,000 nm Spherical Ultrasonication Sonication for 60 min at 20 KHz frequency, NR Gilca et al. 2015
Sarkanda grass 600 W power.
lignin
23 NR Nanocapsules Allyl- 100–400 nm Spherical Miniemulsion; Lignin was grafted with allyl groups via Drug delivery and controlled Chen et al. 2016
functionalized Interfacial crosslinking etherification and dispersed in an oil-in- release of hydrophobic
lignin water miniemulsion by ultrasonication, compounds
and then reacted with a thiol-based
crosslinking agent at the interface of
miniemulsion droplets.
24 NR Nanocapsules Kraft lignin 200–300 nm Hollow spherical Solvent exchange A solution of lignin in ethanol was Potential applications in Li et al. 2016
prepared and the ultra-pure water was various fields
dropped into this solution (90%).
26 PKL Nanoparticle Kraft lignin 19–28 nm Quasi-spherical Anti-solvent Lignin was dissolved in DMF, and the Applications in cosmetics, Myint et al. 2016
nanoparticles precipitation solution was sprayed into the precipitator drug delivery systems, and
using compressed liquid CO2. nanocomposites materials
27 IATNP, HPLNP Nanoparticle Kraft lignin and 45–250 nm Spherical Acid precipitation; IAT lignin dissolved in EG and then HCl Potential application at Richter et al. 2016
Organosolv Solvent exchange was added; HPL dissolve in acetone and extreme pH conditions
then H2O was added.
28 PLA –LNP/ Binary and Pristine lignin NR Film Acid precipitation Lignin dissolved in EG and then HCl was UV light blocking capability, Yang et al. 2016a,
CNC; PVA/ Ternary polymeric added and grafted with glycidyl helpful towards harmful 2016b
CH/LNP films methacrylate and poly (lactic acid). pathogens, helpful in the food
packaging sector
29 LPAM100; Nanoparticle Kraft lignin 5–12 nm Spherical NR By reacting lignin with potassium Application ranging from Silmore et al. 2016
LPAM700 xanthate, 2-bromopropionic acid, and lubrication to improved
thionyl chloride in dry THF under nitrogen composite materials.
at 70 °C overnight.

3
30 NR Nanoparticle LignoBoost™ 200–500 nm Spherical Solvent exchange Kraft lignin was dissolved in THF and Potential application in Lievonen et al. 2016
water was subsequently introduced into composites, Pickering
the system via dialysis. emulsions and antimicrobial
materials. Drug delivery
carrier for cancer therapy
31 LNP Nanoparticle Agave tequilana 1200–3800 nm Spherical Alkaline precipitation Lignin slurry was stirrer and followed by As a skin protector Gutiérrez-
lignin (Organosolv), NaOH, NH4OH, formaldehyde solution was Hernández et al.
400–1400 nm (Soda) added and heating to 85 °C. 2016
32 pLNP; Fe- Nanoparticle LignoBoost™ 200–220; 139–181; Spherical Solvent exchange A mixture containing 50:50 w/w of lignin Promising for cancer therapy Figueiredo et al.
LNPs; Fe3O4- softwood kraft 449–479 nm solution and oleic acid coated Fe3O4 NPs in and diagnosis 2017a
LNPs lignin THF was prepared and dialyzed against
Milli-Q water.
33 CLNPs Nanoparticle LignoBoost™ 218–262 nm Spherical Solvent exchange Lignin was dissolved in THF and water was Targeted and pH-responsive Figueiredo et al.
softwood kraft subsequently introduced into the system delivery of anticancer drugs 2017b
lignin via dialysis
34 QAL/SDBS Micelles Alkali lignin 45–110 nm Spherical Self-assembly Lignin was reacted with CHMAC to prepare Controlled and pH responsive Li et al. 2017
QAL and dialysis in water; QAL mixed with delivery of hydrophobic oral
SDBS, heat and precipitate dissolve in drugs
ethanol.
36 NR Nanoparticle Lignin from NR Spherical Adsorption Lignin suspensions were prepared by Support for enzyme Gong et al. 2017
bamboo shoot suspending the lignin in the buffer and immobilization
shells varying the final concentration.
37 NR Nanoparticle Lignocellulosic 20–100 nm Cubodial Solvent exchange; Cryo crushing of dried fruit; Organo Applications across fields Rangan et al. 2017
fibers Enzyme mediated solvent extraction of powdered fruit, including automobile,
Enzymatic hydrolysis in acidic pH. pharmaceutical and polymer
industries
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Bioresource Technology Reports 9 (2020) 100374
 Table 1 (continued)

S. No. Abbrevation Morphology Raw material Diameter Shape Preparation method Brief description of method Application Reference
P.S. Chauhan

38 LNP Nanoparticle Steam treated 500–1000 nm Spherical Solvent exchange; Steam pre-treatment of lignocellulose, Helpful in integrated Tian et al., 2017a
poplar, pine, corn Enzyme mediated enzyme treated residue extracted with lignocellulose biorefinery
stover DMSO and dialysis with tap water. process
39 NR Nanoparticle/ Cocount fibre 2–150 μm Spherical Microbial hydrolysis Bulk lignin degraded by fungi under Useful in textile, biomedical Juikar and
nanolignin shaking condition followed by and environmental Vigneshwaran,
centrifugation. applications 2017
40 DLNPs/PVA; LNP films Poplar biomass 195 nm; 197 nm Spherical Solvent exchange Lignin were dissolved in DMSO and Nanocomposites having high Tian et al. 2017b
OLNPs/PVA subjected to micellization using dialysis; UV-shielding efficacy,
blend with PVA. antioxidant activity, and in
advanced packaging field.
41 AL/RSV/ Nanoparticle Alkali lignin 131.2 nm Spherical Self-assembly Alkali lignin was dissolved in Methanol, Efficient delivery of Dai et al. 2017
Fe3O4 NPs Ethanol, THF and water was subsequently hydrophobic drug
introduced into the system via dropwise.
42 LNPs Nanoparticle Wiley-milled 108.6–238.9 nm (AL Spherical and Self-assembly Fractionate of poplar wood and spent acid Helpful in bioremediation Chen et al. 2017
poplar NE222 conc. 0.5 mg/ml) Non Spherical liquor stream containing mainly dissolved
lignin that converting into nanoparticle
upon dilution with water.
43 NL Nanoparticle Kraft lignin 10–50 nm Irregular Ultrasonication Lignin dissolved in water and Bioderived fillers for Gonzalez et al. 2017
ultrasonication for 2,4 and 6 h. advanced nanocomposite
applications
44 LNP Nanoparticle Steam exploded 15–20 nm Spherical Acid precipitation SERSL dissolve in EG or castor oil and HCl Acted as a anticorrosive Rahman et al. 2018
Rice straw lignin was added dropwise to the reaction nanofillers for the protection
mixture in a nitrogen atmosphere. of carbon steel
45 LN-g-PAA Nanoparticle Alkali lignin 40–60 nm Uniform and flat Acid precipitation AL dissolved in polyethylene glycol and Used as a adsorbent to Azimvand et al.
HCl was added dropwise to the reaction remove Safranin-O dye and 2018a, 2018b,
mixture until pH 4.0. Basic Red 2 dye 2018c

4
46 CLPs Nanoparticle LignoBoost™ 300 nm Spherical Self-assembly Lignin dissolved in THF and dialysis system To prepare bionanomaterials Mattinen et al.
with water. for advanced applications 2018a, 2018b
80–1000 nm Lignin was dissolved in NaOH under
stirring and then HCl was added.
47 NR Microsphere Commercial lignin 6.85–43.90 μm Spherical Reverse phase Lignin dissolve in ammonia, Potential applications on Yu et al. 2018
polymerization Hexamethylenetetramine and water adsorption or supercapacitor
thereafter formaldehyde was added.
Heated at 94 °C for 2 h in oil phase and
cooled down to get microsphere.
48 L-NPs; L-NPs- Nanoparticle Switch grass lignin 80–800 nm; Spherical Ultrasonication Lignin was dissolve in water, NaOH and Application as a flocculant in Yin et al. 2018
gelatin 500–2000 nm sonication for 60 min at 600 W. Gelatin is wastewater treatment and
attached through direct mixing in different detection of microorganism,
ratio. etc.
49 CLPs; Enzyme Nanoparticle Pine kraft lignin 175–179 nm Spherical Solvent exchange KL was dissolved in THF and water was Application in aqueous ester Sipponen et al.
coated c-CLPs subsequently introduced into the system synthesis 2017; Sipponen
via dialysis. Hydrolase/Lipase is attached et al. 2018
through adsorption on surface.
50 LNP; PMMA-g- Nanoparticle/ Pristine lignin 32.5–65.1 nm Spherical Acid precipitation; Lignin dissolved in EG and then HCl was Application in automotive, Yang et al. 2018a
LNP Nanocomposite Solvent-free radical added. For PMMA-g-LNP, MMA monomer flooring, acrylic glasses and
polymerization, Micro and LNP mixed, ultrasound for 30 min lenses
extrusion followed by benzoyl peroxide was added,
cooled down and heated.
51 PVA/Ch/LNP Nanoparticle/ Pristine lignin 40–60 nm Spherical Acid precipitation; Lignin dissolved in EG and then HCl was Application in drug delivery, Yang et al. 2018b
nanocomposite Freezing-thawing added; For PVA/Ch/LNP, PVA dissolve in food packaging, wound
water and then add LNP. Chitosan dressing
dissolved in 1% acetic acid and mix with
above solution, final mixture freeze thaw
up to 5 cycles.
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Bioresource Technology Reports 9 (2020) 100374
 P.S. Chauhan

Table 1 (continued)

S. No. Abbrevation Morphology Raw material Diameter Shape Preparation method Brief description of method Application Reference

52 LNPs Nanoparticle Alkaline lignin 80–230 nm Spherical Solvent exchange AL dissolved in APS solution and dilute Drug delivery vehicle for Chen et al. 2018
with water up to below MHC to obtain LNP hydrophobic compounds
precipitate.
53 LBNT Nanoparticle Alkaline lignin 154.4–165.6 nm Mostly spherical Inverse-emulsion Lignin dissolved in distilled water, with an Application in bioremediation Xiao et al. 2019
copolymerization amine, and formaldehyde emulsified in and antimicrobial.
liquid paraffin by a surfactant. Heated and
cooled down to 30 °C after adding CS2 for
esterification and freeze drying to obtain
LBNT.
54 LnP-AcL Nanoparticle Alkali lignin 200–800 nm (size can Spherical Solvent exchange; Acetylated lignin dissolves in THF and Application in UV absorbance Wang et al. 2019b
be controlled) Ultrasonication ultrasonication to different intensity. in sunscreen creams.
Precipitates were dialyzed with DDW.
55 LNPs Nanoparticle Soda lignin 50–300 nm Aggregated Solvent exchange/ Soda lignin dissolved in acetone and water, UV-blocking films for Xing et al. 2019

5
particles Nanoprecipitation filtered and filtrate added rapidly in potential applications in
deionized water. Acetone removed and agricultural or food
centrifuged to get LNP. packaging materials.
56 LNPs Nanoparticle Iroko saw dust and NR NR Solvent exchange Lignin dissolved in DMSO and dialysis with Wood-protection applications Zikeli et al. 2019
Norway spruce excess water. against weathering
57 LNPs Nanoparticle Corn stover 132–1099 nm Spherical and Acid precipitation; Fractionated lignin from each SOFA was Application in biorefineries. Liu et al. 2019
uniform Solvent exchange precipitated in HCl, than centrifuged,
washed with distilled water and freeze
dried. The lignin was dissolved in THF,
sonicate it and dialysis with water.
58 NR Nanoparticle Kraft lignin 30–152 nm NR Acid precipitation Lignin (protobind-1000) dissolved in EG, NR Yang et al., 2019
precipitated by HCl.

NR = Not reported. HX: Heteroxylan. IAT: Indulin AT. PDEAEMA: poly(2-(diethylamino) ethyl methacrylate). THF: Tetrahydrofuran. EG: Ethylene glycol. CO2: Carbon dioide. HCL: Hydrochloric acid. HNO3: Nitiric acid.
DMF: Dimethylformamide. NaOH: Sodium hydroxide. NH4OH: Ammonium hydroxide. QAL: quaternized alkali lignin. DMSO: Dimethyl sulfoxide. PVA: Polyvinyl alchol. PMMA: Poly(methyl methacrylate). MMA: Methyl
methacrylate. Ch: Chitosan. CHMAC: 3-chloro-2-hydroxypropyltri methylammonium chloride. SDBS: Sodium dodecyl benzenesulfonate.
Bioresource Technology Reports 9 (2020) 100374
P.S. Chauhan
exchange, ultrasonication, self-assembly, interfacial crosslinking and
emulsion, antisolvent precipitation, microbial and enzyme mediated,
freeze-drying and thermal stabilization, homogenization and alkaline
precipitation has been discovered by which lignin can be converted into
lignin nanoparticles (Table 1).
2.1. Polymerization
In a first of its kind, the coniferyl alcohol was polymerized to syn-
thesize the arabinoxylan-dehydrogenation polymers (synthetic lignin
polymer) nanoparticles. During this process, a combination of coniferyl
alcohol/sinapyl alcohol was used with two structurally-related hetero-
xylans (HX). The nanoparticles produced in this process were char-
acterized by the help of Light Scattering (LS), Atomic Force Microscopy
(AFM), Transmission Electron Microscopy (TEM) and fluorescent
probes (Barakat et al. 2007; Barakat et al. 2008). Qian et al. (2014b)
synthesized N,N-diethylaminoethyl methacrylate (DEAEMA)-grafted
LNPs via Atom transfer radical polymerization (ATRP), which had size
range of 237 to 404 nm. The produced nanoparticles were utilized in
the CO2/N2-switchability pickering emulsions as a surfactant which was
in correlation with graft density as well as the DEAEMA's chain length.
2.2. Acid precipitation
A unique acid preparation method was developed by Frangville
et al. (2012) in order to produce novel LNPs that remain non-toxic for
yeasts as well as microalgae. In this research, two different methods
were compared to synthesize the nanoparticles that have variations in
its stability upon change in pH. In the first method, low-sulfonated
lignin gets precipitated by ethylene glycol solution in the presence of
diluted acidic aqueous solutions. In this way, the LNPs can be retrieved
and their stability is high in different pH ranges. In the second method,
the lignin gets acid-precipitated from high pH aqueous solution thus
resulting in the formation of LNPs that has stability only at acidic range
(Fig. 1).
Gupta et al. (2014) also explored the use of acid precipitation
Fig. 1. Lignin NP synthesis with the proposed mechanism a) the Indulin AT (IAT) getting precipitated from EG with HCl (aq.) and possible subsequent cross-linking
and dialysis, and by b) IAT getting precipitated from aqueous solution in basic range of pH to acidic range. The lignin NPs thus synthesized in this method (b) dissolve
when the pH is above 5 whereas the ones that were synthesized through method (a) can sustain up to pH 10 even in the absence of cross-linking step (Frangville et al.
2012).
6
Bioresource Technology Reports 9 (2020) 100374
technology and synthesized the LNPs. The particle size was measured
using TEM and Dynamic Light Scattering (DLS) and it was observed that
particles do not get aggregated and they are uniformly distributed
which allows to upscaling of this process. Gilca et al. (2014) reported a
chemical method to synthesize nanoparticles using hydroxymethylated
lignin and optimized the conditions for particle size point of view. The
LNPs produced from this process contained high number of hydroxyl
groups due to which they can replace the phenol in the synthesis of
phenolformaldehyde resin, composites, biocides etc. Furthermore,
Richter et al. (2015) and Richter et al. (2016) also synthesized the LNPs
with the precipitation method discussed above, additionally silver ions
were infused in the LNPs and coated with poly(diallyldimethylammo-
nium chloride) (PDAC). Another unique method for the synthesis of
high amount of spherical LNPs was developed by Rahman et al. (2018)
using different media such as Water (W), Ethylene Glycol (EG) and
Castor Oil (CO) through acid precipitation technology and nano-
particles were characterized by TEM, DLS and Scanning Electron Mi-
croscopy (SEM). Azimvand et al. (2018a; 2018b; 2018c) synthesize the
LNPs using alkali lignin as a raw substrate after modification by EG
characterized by DLS, SEM, Fourier-transform infrared spectroscopy
(FTIR), Thermogravimetric analysis (TGA-DTG). Yang et al. (2019) in-
vestigated the role of EG on solution structure of lignin before the re-
duction in pH thus enabling the assessment of the solvent quality, the
conformation of lignin subunits and their aggregation in EG.
2.3. Solvent exchange method
In the solvent shifting method, a solution of an organic compound in
a water miscible organic solvent, which is mixed with an excess of
water, and nanoparticles are generated due to their decreasing solubi-
lity. There is a vast range of application available for this process and it
show a straightforward system. But the major disadvantage in this
process is the amount of low solid content i.e., approximately 1 wt%
(Beisl et al. 2017). For the first time, Qian et al. (2014a), produced the
colloidal lignin spheres of 110 nm based on self-assembly method in
which acetylated lignin was dissolved in Tetrahydrofuran (THF) and
P.S. Chauhan
the water (67%) was added gradually to the solution. This caused the
lignin molecules to associate with one another because of hydrophobic
interactions.
Yearla and Padmasree (2016) study focused on the generation of
spherical dioxane lignin nanoparticles (DLNP) and alkali lignin nano-
particles (ALNP) from two lignin sources such as softwood alkali lignin
(AL) and hardwood dioxane lignin (DL) that was extracted from the
stem region of subabul. Kai et al. (2015) produced lignin nanoparticles
of controllable size using acetylated lignin dissolved in THF and water
was added gradually to the solution. Similarly, Li et al. (2016) prepared
nanocapsules of 10–100 nm range by dissolving the kraft lignin solution
into the ethanol and drop by drop addition of water. The nanocapsules
are preferred for different applications due to its distinct characteristics
such as cheap preparation cost, easy preparation methods and en-
vironment-friendly features. Lievonen et al. (2016) introduced a direct
method for the production of LNPs from the waste lignin obtained from
kraft pulping. When the softwood kraft lignin was dissolved in THF, the
spherical-shaped LNPs were obtained after which the water was in-
troduced in the system via dialysis. There was no need to chemically
modify the lignin in this case… The nanoparticles' final size is decided
on the basis of pre-dialysis concentration of the dissolved lignin. This
study also investigated the firmness of the nanoparticle dispersion in
terms of pH, salt concentration and time. The stability of the lignin
nanoparticle was above 2 months in both pure water as well as in room
temperature, but in case of low pH or high salt concentration, it in-
creased the aggregation of LNPs. Figueiredo et al., 2017a; 2017b) de-
veloped three types of spherical LNPs that had been prepared by solvent
exchange. These three types were pure lignin nanoparticles (pLNPs),
iron(III)-complexed lignin nanoparticles (Fe-LNPs), and iron oxide
(Fe3O4) nanoparticle infused lignin nanoparticles (Fe3O4-LNPs). When
a lignin solution was mixed with oleic acid-coated Fe3O4 nanoparticles
in THF (50:50 w/w) followed by a water dialysis, it resulted in the
formation of Fe3O4-LNPs. Since it possesses excellent paramagnetic
behaviour, it is considered as the best choice for diagnosis and treat-
ment of cancer. Tian et al. (2017a) developed a thin film through the
amalgamation of polyvinyl alcohol with dual kinds of LNPs namely
DLNP and ALNP on the basis of lignin isolation technical method. To
achieve full utilization of lignocellulosic biomass and easy integration
of LNPs production into current biorefinery concept, a two-step pre-
treatment strategy, mild steam pretreatment followed by solvent ex-
traction, was employed to produce Deep eutectic solvent (DES) and
organosolv technical lignins from raw hardwood poplar. Both the
technical lignin dissolved in dimethylsulfoxide (DMSO) and underwent
micellization with the help of dialysis process resulting in the produc-
tion of evenly distributed LNPs often mentioned as DLNPs and OLNPs,
respectively. The LNPs thus produced were spherical in shape with
novel core-shell nanostructure.
Sipponen et al., 2017, 2018) synthesized Colloidal Lignin Particles
(CLPs) by dissolving the softwood kraft lignin in THF: water ratio of
5:1 v/v over a period of 3 h and then precipitating by rapid pouring of
the solution into 380 g of vigorously stirred deionized water. Various
weight proportions of cationic lignin were then coated onto the CLPs in
soluble fraction. In comparison, the cationic CLPs were able to stabilize
a wide range of long-lasting pickering emulsions than its counterpart
i.e., irregular kraft lignin particles or otherwise the regular CLPs. Wang
et al. (2019b) proposed a very simple, yet unique and green technique
to produce regular LNPs of high quantity. Lignin was first modified
through a microwave acetylation process without any catalysts and
solvents other than acetic anhydride, which acted as both reaction re-
agent and dispersion solvent. Consequently, high yielding regular LNPs
were produced through a combination of solvent shifting and ultra-
sound process. The produced LNPs can be easily separated through
centrifugation and used THF can be recycled and reused which simply
reduce the cost of the process. Interestingly, upon increasing the con-
centration of lignin content and ultrasound intensity, yield of LNPs has
also been increased. Xing et al. (2019) produced LNPs by dissolving
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Bioresource Technology Reports 9 (2020) 100374
soda lignin in acetone and water, filtered and filtrate added rapidly in
deionized water. Through rotary evaporation, acetone was removed
after which the solution was centrifuged to obtain LNPs. For the first
time, the study implemented a bioinspired melanin-like polydopamine
thin layer in the LNPs which increases their biocompatibility. Recently
Zikeli et al. (2019) separated the lignin from wood wastes which con-
sisted of Iroko sawdust (IR) as well as mixed sawdust from Iroko and
Norway Spruce (IRNS) and nanoparticles were produced via solvent
exchange method by first dissolving lignin in DMSO and dialysis with
excess water.
2.4. Ultrasonication
Dry and wet milling techniques as well as other such mechanical
treatments are generally utilized to reduce the particle size to nano-
metre scale. However, it has few disadvantages such as non-uniformity
in the size and broad particle size distributions. In spite of these, it is
still preferred because it is a simple method to produce nanoparticles
(Beisl et al. 2017). Gilca et al. (2015) used the sonication process in
order to reduce the size of lignin extracted from wheat straw and sar-
kanda grass. The synthesized nanoparticles were in the range of 10 to
50 nm and they were characterized both morphologically and di-
mensionally to evaluate any abnormal changes in its composition and
structure. Results confirmed that there was no significant relationship
between the applied intensity and the compositional and structural
changes of nanoparticles. This is an even more attracting method for
future applications since none of the solvents used in this process are
hazardous. Gonzalez et al. (2017) produced LNPs by the ultrasonic
treatment for different time periods and oscillation amplitude was 95%
in order to get the lignin-water dispersions having superior colloidal
stability. Yin et al. (2018) also produced LNPs through the ultrasonic-
assisted alkali method using switch grass as a lignin source by dissol-
ving into sodium hydroxide and then sonicated at about 600 W for 1 h
to obtain LNPs of 220 nm average size.
2.5. Self-assembly
Self-assembly is a process in which an incoherent system of the pre-
existing components generates a pattern or an organized structure be-
cause of the specific interactions in the absence of any external direc-
tion. A self-assembly method was made use for the preparation of lignin
reverse micelles (Qian et al. 2015a). In this method, cyclohexane was
added into alkali lignin solution that was pre-dissolved in dioxane. With
further addition of cyclohexane, there may be possible flocculation and
precipitation of lignin micelles occur. The size of the nanoparticles
rendered, was the range of 120 nm to few micrometers on the basis of
cyclohexane content. Li et al. (2017) proposed an easy and practical
method in the preparation of lignin-based complex spherical micelles in
green solvents that possess pH-responsive features. After quaterniza-
tion, the alkali lignin (AL) was left to self-assembly so as to form the
lignin-based complex micelles along with sodium dodecyl benzene-
sulfonate (SDBS) in ethanol/water mixture with increasing amount of
water i.e. 0 to above 86%.
Dai et al. (2017) created a first-of-its-kind green nanoparticle plat-
form using lignin in the absence of chemical alterations. The corn cob
was utilized as a source of AL in the preparation of spherical-shaped
nanoparticles that possess good dispersion characteristic. In this pre-
paration process, a simple self-assembly method was followed in which
the water was added with methanol solution of AL. Mattinen et al.
(2018a, b) prepared small-sized colloidal lignin particles (CLPs) in-
cluding bilayer polypeptide alterations from KL utilizing self-assembly
so as to make the CNF surfaces adapt to the heat treatment. Liu et al.
(2019) produced LNPs on the basis of Sequential Organosolv Frag-
mentation Approach (SOFA) using ethanol as well as various stages of
catalysts were explored for selective-dissolution of lignin from corn
stover. This was performed to generate multiple uniform lignin streams
P.S. Chauhan
and alter its chemical characteristics as well as its reactivity so that the
LNPs can be fabricated using expected quality features through self-
assembly. The effective diameter more or less was in the footsteps of
stage 1 (50% ethanol +1% sulphuric acid), stage 3 (50% ethanol), and
stage 2 (50% ethanol +1% formic acid) in every SOFA. The smallest
effective diameter was 130 nm approximately from SOFA that utilized
ethanol as well as sulphuric acid.
2.6. Interfacial cross linking and emulsion
Emulsification and cross linking are the two phenomena in asso-
ciation with ultrasound to produce lignin microcapsule. It was observed
lignin cross linking capability was induced at water/oil interface and
enhanced by addition of cross linking agent. Mechanism involve in this
process comprise two steps: the first process depends on the collapse
intensity of the cavitation bubbles (responsible for the shear forces).
The oil-in-water emulsion was obtained applying an acoustic power of
160 Wcm−2 for 40 s. The amphiphilic lignin chains are thought to
diffuse towards the oil microdroplets so as to stabilize the water/oil
interface. In the second step, lignin chains is induced for cross linking
by the hydroxyl (•OH) and superoxide (HO2 − •) radicals generated
during the acoustic cavitation process (Tortora et al. 2014). Nypelo
et al. (2015) prepared spherical particles using a water-in-oil micro-
emulsion and thereby mixing of lignin solution with a blend of sur-
factants, for instance, Span 80, Tween 80 and pentanol pre-dissolved in
octane. This led to the formation of hydrophilic–lipophilic balance with
ca. 7. In the another study performed by Chen et al. (2016), lignin
nanocapsules having pH-responsive features has been synthesized
through interfacial miniemulsion polymerization followed by cross-
linking. In the first step, the ultrasonication process was performed to
obtain an oil-in-water miniemulsion by mixing vigorously butyl acetate,
hexadecane (co-stabilizer), azobisisobutyronitrile (AIBN, oil soluble
initiator) and the cross-linker, trimethylolpropane tris(3-mercaptopro-
pionate) and further mixed with water phase bound lignosulfonate
[with and without sodium dodecyl sulfate (SDS)]. After this process, a
cross-linking agent was made to undergo reaction with lignin at mini-
emulsion droplet interface so as to produce the nanocapsules. The
prepared lignin nanocapsule possesses pH responsive properties due to
presence of acid-labile b-thiopropionate cross-linkages in the capsule
shell.
Xiao et al. (2019) developed a Lignin-Based Nano-Trap (LBNT)
using a simple inverse-emulsion copolymerization method. Initially
lignin was dissolved in the distilled water using amine and a cross-
linker (CH2O), it was then emulsified in liquid paraffin using a surfac-
tant (Span80) followed by heating. This was then cooled down to 30 °C
to which CS2 was added so that it undergoes esterification. The solution
was then centrifuged, washed and freeze-dried so as to finally obtain
the LGNT. The following features are expected in LGNT reduced sized of
the particles so that the diffusion and contacting frequency can be in-
creased; further it should possess heavy metal-binding groups on its
surface that can afford various heavy metal ions with good adsorption
capability and finally the surface-dispersed binding sites should be
controlling the heavy metal ions thus helping in various applications.
2.7. Antisolvent precipitation
Over the last two decades, the “compressed/supercritical fluid”
based technology used in the production of polymeric nanoparticles has
gained considerable attention. Its capability to control the size and size
distribution as well as morphology are its unique features which are
critical in the development of pharmaceutical and drug delivery ap-
plications. CO2 gained special attention since it possesses excellent and
critically-important features (TC = 304.3 K and PC = 7.4 MPa) because
of its physicochemical properties can be fine-tuned and its cheap, non-
toxic, non-flammable with abundant quantities. In addition to the
above, it remains a poor solvent in case of macromolecules, namely
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Bioresource Technology Reports 9 (2020) 100374
polymers, due to which it is often cited as the best antisolvent for
precipitation processes in which the precipitates can be influenced
through the control of temperature and pressure (Beisl et al. 2017). Lu
et al. (2012) made use of 80.5% pure poplar OS in a supercritical CO2
(SC-CO2) process. In this method, precipitation chamber was filled with
SC-CO2 at 30 MPa under 35 °C temperature with pure acetone. After
reaching stable conditions, 0.5 g/L lignin/acetone solution was used to
replacing the acetone and precipitated nano-scale particles were re-
moved out of the stainless steel-made frit in precipitation chamber.
Result shows upon conversion into nano scale lignin solublility were
significantly enhanced in the water. Myint et al., (2016) produced LNPs
using compressed fluid anti-solvent method in which nanoparticles
were quasi-spherical and identical in nature. In this method, firstly KL
was dissolved in DMF after which it was sprayed onto the precipitator
with the help of compressed liquid CO2 (clCO2) as an anti-solvent. This
led to the formation of identical and quasi-spherical LNPs which shows
enhanced solubility and stability in water in comparison to raw lignin.
2.8. Biological method
Lignocellulose, the most abundant renewable biomass on earth,
chiefly consists of cellulose, hemicelluloses, lignin and pectin as the
major constituents with resins, waxes, proteins and extractives as minor
constituents. Recently, there is an increased potential tapped in the
enzymatic methods followed in biomass degradation processes by
combining saccharification and fermentation processes. When the
lignin is converted into nanolignin through enzymatic hydrolytic pro-
cess, especially, with the help of lignocellulosic materials, there is an
improvement observed in the properties of the materials. For the first
time, Rangan et al. (2017) prepared lignin-rich cubodial nanoparticles
from lignocellulosic fibers generated from Indian ridge gourd (Luffa
cylindrica) through the breakdown of lignin-cellulose complex using
specific enzymes. After this Juikar and Vigneshwaran, 2017 generated
LNPs using coconut fibers as sources with the help of Aspergillus sp., a
fungal isolate that is capable of producing high amount of lignin per-
oxidase enzyme. The bulk lignin size was between 2 and 150 μm with
the peak value being 55 μm. On comparison between the microbial
processes and homogenization & ultrasonication processes, the yield of
nanolignin in the former was 58.4% whereas it was 81.4% and 64.3% in
the latter respectively. Following this study, Tian et al., (2017b) pro-
duced LNPs from cellulolytic enzyme treated lignin (CEL), obtained
from steam-pretreated, agriculture reside corn stover, hardwood po-
plar, and softwood lodge pole following the prevalent dialysis method.
In this method, the LNPs produced were in the range of 81.8%, 90.9%
and 41.0% with their relevant average particle sizes being 218, 131,
and 104 nm, respectively. The high-quality LNPs thus obtained were
spherical in shape with abundant functional groups and high stability in
the pH range of 4–10, which showed tremendous promise for the ap-
plications in the emerging nanomaterial fields (Fig. 2).
2.9. Freeze-drying and thermal stabilization
Gonugunta et al. (2012) synthesized LNPs through combination of
freeze-drying process, thermal stabilization as well as carbonization
processes using lignin as a renewable source. The researchers in-
vestigated the effect of adding different amounts of KOH to lignin so-
lution in terms of its solubility, freeze-drying process and thermal sta-
bilization of the freeze-dried lignin.. The SEM analyses confirmed lignin
was influenced to form porous microstructure due to freeze-drying
process. Furthermore, TEM analysis showed that the thermal stabili-
zation of freeze-dried lignin has stopped the generation of agglomer-
ated carbon nanoparticles at the time of carbonization. The carbon
nanoparticles had the least size range of 25 nm when prepared using
lignin precursor and 15% KOH.
P.S. Chauhan
Fig. 2. Production of integrated lignin nanoparticles into lignocellulosic biorefinery – a diagrammatic representation. The enzymatic hydrolysis was performed over a
period of 72 h at 2% w/v solid loading using Cellic Ctec3. The lignin was extracted in a very mild condition (2% w/v, 3 h, 80 °C) using DMSO before subsequent
dialysis at room temperature. Then the dialysis was stoped until all the DMSO is evaporated from the wastewater (Tian et al., 2017a).
2.10. Homogenization
Nair et al. (2014) prepared nanolignin with the help of a simple high
shear homogenizer. The kraft lignin particles with a broad distribution
ranging from large micron- to nano-sized articles were completely
homogenized to nanolignin particles with sizes < 100 nm after 4 h of
mechanical shearing. Based on the results obtained from 13C nuclear
magnetic resonance (NMR) and 31P NMR analyses, it was found that
there was no significant chemical composition changes occurred in
kraft lignin particles in comparison with mechanically-treated nano-
lignin particles. There was no notable change observed among the na-
nolignin particles in terms of molecular weight distribution and poly-
dispersity in comparison with pure lignin particles.
2.11. Alkaline precipitation
Gutiérrez-Hernández et al. (2016) generated LNP via alkaline pre-
cipitation method using lignin obtained from Agave tequilana Weber
bagasse by soda and organosolv. In brief, lignin slurry 17 wt% was
stirred for 1 h followed by the addition of NaOH (exact amount can be
calculated on the basis of lignin content). After 2 h, ammonium hy-
droxide was added and then high intensity mixing was performed using
Ultra-turrax (IKA, T10) at 24,000 rpm for 5 min. Then active for-
maldehyde was added and increased the temperature up to 85 °C for
2 h. Finally, suspension was kept under magnetic stirring at 600 rpm for
the crosslinking and the formation of nanoparticles. Total six types of
lignin NPs were prepared, three for each type of lignin source, using 18
and 27 wt% of active formaldehyde (CH2O) and the lignin samples in
absence of formaldehyde as blanks. L1 (without formaldehyde or
blank), L2 (18 wt% CH2O) and L3 (27 wt% CH2O) are the labels for
every type of lignin source i.e. organosolv or soda.
3. Application of lignin nanoparticles
LNPs have attracted a great deal of attention in the last decade
because of their biotechnological potential in various industrial pro-
cesses. The following section will discuss some of the most promising
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Bioresource Technology Reports 9 (2020) 100374
and newly explored applications of LNPs.
3.1. Antibacterial agent
As research in current medical and biological fields advances, an
increasing interest in the possibility of lignin to act as an antimicrobial
agent has been garnered. The phenolic components present in the lignin
plays the vital role behind the antimicrobial characteristic of the lignin,
more especially the side-chain structure as well as its functional groups.
In general, presence of double bond in α and β positions of the side
chain with a methyl group in γ position in phenolic fragment shows the
highest efficacy against microbes. Gregorová et al. (2011) conducted
detailed investigations about the application of lignin as an anti-
microbial agent through agar diffusion tests for Gram +ve and Gram
-ve bacteria. The antibacterial effectiveness of Bjorkman lignin that was
produced by beechwood was compared with standard additives, for
instance, silver nitrate, chlorhexidine, benzalkonium chloride and
bronopol. Result showed that there were similarities in the diameter of
the growth inhibition for Bjorkman lignin and that of bronopol and
chlorhexidine with both bacterial strains. So, it has been reported that
the lignin's antibacterial property is in par with the existing anti-
microbial agents. Further, this study also investigated the possibilities
of lignin being used as a commercial antimicrobial agent by in-
corporating Bjorkman lignin in polyethylene films, and mechanical
testing was performed which shows that there was no drastic drop in
the mechanical properties. Thus, it confirms that the Bjorkman lignin
can potentially be used in polyethylene films while preserving the an-
tibacterial properties of silver. Another study conducted with the same
objective which shows lignin at a nano scale seems to be useful in the
treatment of dangerous microbes after introducing it into silver nano-
particles. In spite of the fact that AgNPs are applied against a range of
microbes, such as, but not limited to, Escherichia coli and Pseudomonas
aeruginosa, they still are loaded with demerits. In their activated form,
the unused silver nanoparticles remain in the environment and could
adversely affect the ecosystems. Richter et al. (2015) suggested that
lignin as the best choice to replace silver core since the former is
abundantly available with environment-friendly characteristics. They
P.S. Chauhan Bioresource Technology Reports 9 (2020) 100374

Fig. 3. An illustrative presentation with the principle of bactericidal action performed by environmentally-benign lignin-core nanoparticles (EbNPs) and currently
used silver nanoparticles (AgNPs). a, the common phenomenon in the antimicrobial action of AgNPs through the release of Ag+ ions, which continues post-
utilization. b, Antimicrobial action mechanism of Ag+ ion-infused EbNPs with a cationic polyelectrolyte coating which help the electrostatic attraction between
EbNPs and negatively charged cell walls. When compared to AgNPs, there were no Ag + ions present in EbNPs when applied due to which its actions are minimum in
post-utilization activity. c, TEM micrograph of as-synthesized EbNPs sized in the range of 40–70 nm. d, Confocal microscopy image of EbNPs with polyelectrolyte
coating adhering to E. coli cell membrane (Richter et al. 2015).

tested the capability of environmentally-benign LNPs against a range of
bacterial strains so that its antimicrobial efficiency can be evaluated. In
this study bacteria were exposed to LNPs and then controlled the same
for a specific time period before culturing on Luria-Bertani agar plates.
The used controls included polydiallyldimethylammonium chloride
polyelectrolyte solution, silver nitrate (AgNO3) solution, positively
charged branched polyethyleneimine-coated silver nanoparticles, lignin
nanoparticles without silver ions and centrifuged supernatant solution
from the sample. Surprisingly outstanding performance of LNPs was
recorded over controls, AgNO3, and traditional AgNP which had > 12-
times and 20-times excess silver ions respectively. Further, it was also
found that the capability against microbes got lost after the silver ions
got exhausted. But there is still absence of data on minimum con-
centration of lignin needed to get rid of bacteria and its kinetics in
targeting the bacteria. It is crucial to explore the life span of lignin and
its degradation rate so that the amount of substance required to inhibit
the growth of microbes can be found which eventually results in the
elimination of bacteria (Fig. 3).
Interestingly, Yang et al. (2016a) produced ternary polymeric films
on the basis of content of LNPs (1 wt% and 3 wt%) with dispersing
cellulose nanocrystals (CNC) in neat poly (lactic acid) in as well as in
glycidyl methacrylate (GMA) grafted PLA (g-PLA) respectively which
shows excellent antibacterial activity in order to reduce the expansion
of Pseudomonas syringae pv. tomato (Pst), a bacterial plant pathogen.
Furthermore, binary and ternary polymeric films produced on the basis
of polyvinyl alcohol (PVA), chitosan (CH) and lignin nanoparticles
(LNP) using solvent casting method. The capabilities to prevent the
growth of Gram negative bacteria, Erwinia carotovorasub sp. carotovora
and Xanthomonas arboricola pv. pruni over the time was revealed
through antimicrobial assays (Yang et al. 2016b). The Polyvinyl al-
cohol/chitosan (PVA/Ch) hydrogels consist of 1 wt% as well as 3 wt%
of LNPs were also prepared by the same group via freezing-thaw pro-
cedure. The results shows LNPs are effective against Gram -ve bacteria
(E. coli) in comparison with Gram +ve (Staphylococcus aureus and S.
epidermidis) strains when tested using antimicrobial tests. These results
pointed out how this green functionality is able to develop novel stra-
tegies to save from harmful pathogens and how far it is helpful in food
packaging sector (Yang et al. 2018b).
3.2. Antioxidant agent
As mentioned earlier, due to complex chemical structure and pre-
sence of functional groups in lignin, the oxidation propagation reaction
can be stopped through hydrogen donation. Lu et al. (2012) developed
highly concentrated solution of nanolignin (12. 4 times higher than the
raw lignin) which possesses higher antioxidant as well as free radical
scavenging activities, resulting in improved hydrogenating power. Ge
et al., 2014conducted an investigation of nanolignin prepared by al-
kaline solution precipitation method for Free Radical Scavenging (FRS)
activity and it was observed that nanolignin shows 3.3 fold higher

10
P.S. Chauhan
activity compared to control because of small size, higher surface area
and bioavailiability. The results achieved from the 2,2-Diphenyl-1-pi-
crylhydrazyl (DPPH) antioxidant assay inferred that the high amount of
antioxidant activity is present in nanolignin. The IC50 value of na-
noscale lignin was 2.70 ± 0.17 mg/ml indicating that it possesses high
amount of antioxidant activity whereas in case of microscale lignin, the
value as merely 32.21 ± 0.1 mg/ml. Yearla and Padmasree (2016)
performed study in which the dioxane LNPs were fabricated and higher
antioxidant properties were observed than the bulk lignin based on the
data recorded for E. coli survival rate. Domenek et al. (2013) in-
vestigated the antioxidation effect in material applications with the
suggestion that lignin can replace synthetic antioxidants especially in
food protection i.e., integration of active packaging materials with
natural antioxidants. Pouteau et al. (2003) evaluated the polypropylene
(PP)/lignin blends' antioxidant activities on the basis of oxidation In-
duction Time (OIT) with the help of TGA. Results show in the absence
of lignin OIT of PP was 30 min but upon addition to 1 wt% lignin, the
OIT of PP/lignin blends ranging from 70 to 670 min. Furthermore,
when the phenolic content and OIT are compared (in the ratio of ali-
phatic hydroxyl groups to phenolic organosolv lignin groups), the OIT
got increased with the diminishing values of phenolic content. This
denotes the fact that when there is low phenolic content present, it
would increase the compatibility between lignin and PP and accord-
ingly the antioxidant activity too. Morandim-Giannetti et al. (2012)
also performed similar study of lignin in PP/coir fibre (PP/CF) com-
posite showing significant increase in OIT was observed upon addition
of lignin and it vary from content of lignin. Tian et al. (2017a) in-
troduced LNP into conventional polymeric matrix such as poly(vinyl
alchol) to form transparent films which exhibit excellent antioxidant
functionalities (reached ~160 μm mol Trolox g−1 with 4 wt% of LNPs).
Simultaneously, high number of phenolic hydroxyl groups present in
the shell region of the LNPs enabled an excellent interfacial adhesion
with PVA matrix via the formation of hydrogen bonding network which
further improved the mechanical and thermal performances of the
fabricated LNPs/PVA nanocomposite films. The antioxidant properties
and high dissolubility were found in nanolignin. In spite of the fact that
highly-concentrated lignin can exhibit strong anti-oxidant properties
but still there are some serious impacts caused by it. So, there must be
an optimization of lignin concentration in biomaterials to equalize the
cytotoxicity as well as antioxidant activity (Fig. 4).
3.3. UV absorbents
Lignin has tremendous potential in sun block due to its excellent
non-oxidizability capability. Various lignin-based sunscreens were
prepared through the amalgamation of LNPs and pure cream. It was
observed that that the SPF value of pure cream was 1.03 whereas
creams contained different concentration of LNPs has a SPF in the range
Fig. 4. Synthetic fabrication of LNPs and LNPs/PVA composite film. Proposed mechanism for antioxidant activity with the help of LNPs as the functional additive
(Tian et al. 2017b).
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Bioresource Technology Reports 9 (2020) 100374
of 1.26 to 2.23. There was an enhancement in SPF value to 2.23 with
5% increase in LNP content. The performance of the sunscreen cream
having NAcL-2 was better with 2.23 SPF value i.e., 115% increase in
comparison with pure cream. This phenomenon has a direct relation
with LNP size i.e., smaller the size of LNPs, higher the performance of
the sunscreens. The probable reason for the considerable higher per-
formance of the sunscreen with LNPs might be attributed to the con-
jugated system in lignin which might have been produced at the pre-
paration process of the nanoparticles. Furthermore, this system is
supported by easy UV-absorbing functional groups namely methoxyl
groups and S- and G-type lignin in a significant manner. The presence of
π − π stacking between the sunscreen cream and aromatic rings in
LNPs also benefit the performance of the sunscreen creams (Wang et al.
2019b). Tian et al. (2017b) incorporated the nanoparticles in conven-
tional polymeric matrix namely poly(vinyl alcohol) and resultant
transparent nanocomposite film have an added UV-shielding efficacy
(reached ~80% at 400 nm with 4 wt% of LNPs). Gutiérrez-Hernández
et al. (2016) evaluate the photoprotection characteristics of LNPs pro-
duced from Agave tequilana lignin in association with ZnO nanoparticles
(primary active ingredients for sunscreens). The SPF value was in the
range of 4–13 achieved from the mixture of all the components in the
presence of different concentration of LNPs. It was observed that pre-
sence of LNPs were able to absorb both UV-B as well as UV-C regions
which improve the SPF value of sunscreens in comparison to without
LNPs. Xing et al. (2019) prepared the poly(butylene adipate-coter-
ephthalate) (PBAT) films in combination with LNPs, lignin−melanin
core−shell nanoparticles (LMNP) and melanin core - shell nano-
particles (MNP). All the films showed improved tensile properties,
thermal stability with noteworthy UV-blocking ability (> 80% UV-B) in
the presence of NPs concentration ranging between 0.5 and 5.0 wt%.
Yang et al. (2015c) also developed a thin film of poly (lacticacid)
(PLA)/LNP films by premixing 1%wt of LNP into PLA or glycidyl me-
thacrylate(GMA) grafted PLA (g-PLA) with the help of novel master
batch procedure and investigated for accelerated UV weathering (up to
480 h). Results retrieved from UV–vis characterization showed the
better behaviour of the LNPs as a UV light barrier in grafted PLA and it
has been also proved that GMA grafting onto PLA matrix promoted the
dispersion of LNP in matrix. Pucciariello et al. (2008) blended the
biodegradable poly(ε-caprolactone) (PCL) and steam-explosion lignin
by high-energy ball milling process. The lignin addition strongly en-
hanced the UV stabilization of PCL constituents, since the lignin has
phenolic groups which acted as a radical scavenger, thereby reduces or
promotes the rate of radical degradation process. Qian et al. (2015b)
assessed the lignin in the upgradation of high-performing broad-spec-
trums sunscreens. The addition of 2 wt% and 10 wt% lignin leads to
increment of SPF to 30 and 50 respectively from 15. Interestingly,
sunscreen performance improves with UV-radiation time. After 2 h of
UV radiation, the UV absorbance of the 10 wt% lignin SPF 15 lotion
P.S. Chauhan
increases dramatically.. This might be due to the antioxidant property
of lignin as well as specific synergistic effects that present between
lignin and other such components in the lotions. The lignin is con-
sidered as a green option that can replace the synthetically prepared
sunscreen actives. Yu et al. (2015) developed lignin-grafted co-poly-
mers, namely lignin-graft-poly(methyl methacrylate-co-butyl acrylate)
(lignin-g-P(MMA-co-BA) through the process of grafting from Atom
Transfer Radical Polymerization (ATRP) with help from lignin-based
macroinitiators and determined as Sustainable Thermoplastic Elasto-
mers (TPEs). Based on the results, the TPEs lignin-g-P(MMA-co-BA)
copolymers' mechanical properties were improved in a significant
manner compared with linear P(MMA-co-BA) copolymer counterparts
with 70% elastic strain recovery. High amount of absorption was ex-
hibited by lignin-g-P(MMA-co-BA) copolymers in UV spectrum which
might allow the applications in UV-blocking coatings.
3.4. Hybrid nanocomposites
LNPs have also been exploited as reinforcing agents in polymer
matrix and nanocomposites. The resultant copolymers possess excellent
mechanical, thermal properties and biocompatibility compared to ori-
ginal polymers (Feldman 2016; Figueiredo et al. 2018; Yang et al.,
2019. Gupta et al. (2015) made use of LNPs as reinforcing agent to bio-
poly(trimethylene terephthalate) (bio-PPT) hybrid nanocomposites
which had 1.5% weight of LNPs and 7.0% weight of vapor-grown
carbon fibers (VGCF). Results established that there was an enhance-
ment in the mechanical features of nanocomposites, for instance, im-
pact strength in the thermal properties, tensile strength, tensile mod-
ulus and biodegradation characteristics compared to bio-PTT.
LNPs were also utilized as reinforcing agent for those materials
which contain porous structures as shown by Del Saz-Orozco et al.
(2012). In this study LNP's in their different weight fraction were added
into phenolic foams and results were analyzed using analysis of var-
iance approach (ANOVA). The results showed that low density and
smaller cell sizes were obtained, due to the surfactant effect of lignin in
the formulation mixture. In addition, there was a 28% increment in
compressive modulus and 74% increase in compressive strength ex-
perienced was observed in comparison with non-reinforced foams. Qian
et al. (2015a) developed lignin reverse micelles (LRM) films using cy-
clohexane, alkali lignin (AL)/dioxane solution into high density poly-
ethylene (HDPE). It was observed that water contact angle of the LRM
films was > 80° while it was only 52° in AL film control sample this
leads to an improvement in the miscibility LRM films because of hy-
drophobic nature. There was also an increase in mechanical strength as
well as young's modules from 1066 to 2104 MPa when 5 wt% LRM was
loaded.
Yang et al. (2015a) incorporated 0, 1 and 3%wt of LNPs into Wheat
Gluten (WG) bio-nanocomposites in order to efficient absorbance of the
UV spectrum and enhancement of the water sensitivity. Further, in spite
of the fact that when LNPs are added, it reduces the transparency at two
varied weight contents, but it still improved the thermal stability, glass
transition as well as the mechanical behaviour of the WG-based bio-
nanocomposites. In addition to the above, Yang et al. (2015b) prepared
LNPs/polylactic (acid) (PLA) bio-nanocomposites through the processes
of melt extrusion (E-PLA) and solvent casting (C-PLA) with an aim to
increase the content of lignin. The nucleation effect was proved to be
effectively improved at the time of accomplishing homogenous dis-
persion of LNPs in PLA matrix at 1 wt% in melt-extruded samples but on
further addition of LNPs, there was no favourable crystallization be-
haviour observed. Silmore et al. (2016) developed polymer-grafted
lignin nanoparticles (PGLNs) with the help of reversible addition-frag-
mentation chain transfer (RAFT) chemistry in order to tune the ag-
gregation strength while at the same time it also retain the interfacial
activities in generating the pickering emulsions. The results observed,
notified a fact that there is a significant impact of salt concentration on
zeta potential, aggregation, as well as interfacial tension. These factors
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Bioresource Technology Reports 9 (2020) 100374
are responsible for the alterations in solubility of both kraft lignin as
well as polyacrylamide grafts.
Sun et al. (2015) prepared PLA-lignin composites by mixing lignin-
g-rubber-g-poly (D-lactide) copolymer particles and commercial poly (L-
lactide) (PLLA) in chloroform. Although this method observed as an
efficient for the dispersion of lignin in PLA matrix but it is challenging
and cost-consuming. In addition to the above, the lignin was modified
by Chung et al. (2013) through graft copolymerization reaction using
lactide and in the presence of triazabicyclodecene (TBD) as a catalyst.
By changing the lignin/lactide ratio and pre-acylation treatment, one
can easily control the PLA's chain length in lignin-g-poly (lactic acid)
copolymer. It was observed that there is a notable increase in the tensile
strength as well as strain up to 16% and 9% respectively upon addition
of lignin-g-PLA copolymers without losing tensile modulus.
Jiang et al. (2014) suggested the dual role of lignin as a cross-linker
to cure epoxidized natural rubber (ENR) as well as reinforcing filler to
enhance the mechanical properties of the lignin/ENR composites. It was
hypothesized that presence of hydroxyl and carboxyl groups of lignin
could react with ENR's epoxy sites through ring opening reaction under
a high-temperature dynamic heat treatment (180 °C) leads to an even
dispersion of lignin along the matrix of ENR and provides excellent
compatibility to rubber materials. Liu et al. (2015) developed compo-
site fibers with lignin/PAN/carbon nanotubes (CNT) following the gel-
spinning technology. The PAN, PAN/lignin and PAN/lignin/CNT were
transformed into carbon filters under carbonization (1100 °C) and
identical stabilization conditions.
Yang et al. (2014) prepared lignin-based xerogel having excellent
self-cleaning properties and super hydrophobicity with the use of
lignin, modified diisocyanate (MMDI) and polyurethane. The produced
xerogel capable to absorb high amount of oil from the oil/water mix-
ture which open the new route in removal of spilled oil. Few years after
Mahmood et al., 2016prepared polyurethane based lignin foam
showing enhanced mechanical properties, thermal stability, curing
rates, and also the flame resistance in comparison to without lignin
based polyurethane foam, which suggest that presence of lignin is ad-
vantageous in foam and rubber materials (Hatakeyama et al. 2005).
Peng and Chen (2011) developed pH-sensitive hydrogels by the che-
mical crosslinking of acetic acid lignin with isocyanate group-termi-
nated polyurethane monomers, in which the swelling ratios enhanced
with the increase of the pH. Moreover, thermal stabilities of the gen-
erated hydrogels were improved and the release profiles of ammonium
sulfate suggested that prepared hydrogels could be used as coating
materials to control the release of fertilizer for different agricultural and
horticultural applications (Peng and Chen, 2011). Yang et al. (2018a)
developed LNPs reinforced Poly(methyl methacrylate) (PMMA) nano-
composites through a combination of hot-press methods, solvent-free
radical polymerization and micro extrusion following the masterbatch
approach. Result shows there was significant improvement was ob-
served in various parameters, specifically, excellent thermal, scratch,
UV resistance and hardness in case of PMMA/LNP nanocomposites and
it also paves the way for these systems to be used in other sectors such
as lenses, acrylic glasses, flooring and automotive industry.
3.5. Drug delivery system
Lignin has the potential to produce nanoparticles for encapsulation
of different compounds for different pharmaceutical applications such
as anticancer therapy (Figueiredo et al. 2017a, 2017b). For the first
time Frangville et al. (2012) explored the use of nontoxic LNPs for
encapsulation of hydrophilic compounds i.e. Rhodamine 6G with high
loading capacity. Thereafter Tortora et al. (2014) prepared hydro-
phobic compound namely Coumarin-6 loaded lignin microcapsule
(LMC) and showed products had low cytotoxicity and could be effec-
tively internalized into Chinese hamster ovary cells. The coumarin-6
release in the presence of SDS solution 5% w/v showed that almost
100% of entrapped Coumarin-6 was released in 60 min. There may be
P.S. Chauhan
two concomitant reasons associated with the release of entrapped
Coumarin-6 in the presence of SDS such as deterioration on lignin hy-
drophobic interactions and the high affinity of the molecule for the new
dispersion fluid. The SDS molecule interfere with the lignin's stabilizing
characteristics at the core−shell boundaries which induce the release
of Coumarin-6 because of the high solubility of the hydrophobic mo-
lecule in SDS solution. From these results, it can be understood that
lignin micro/nanocapsules can be used as the topical applications since
the pool of antioxidants and essential oils may get released if they
contact with lipid tissue or dermatological diseases. One step further
Chen et al. (2016) developed pH sensitive nanocapsule loaded with
hydrophobic coumarin-6, in which release of coumarin-6 can be con-
trolled by altering the pH of outer solution because of the presence of
acid-labile β-thiopropionate cross-linkages in capsule shell.
Figueiredo et al. (2017a) synthesized three different type of LNPs
namely: pLNPs, Fe-LNPs and Fe3O4-LNPs having limited cytotoxicity
with different tested cell lines and only 12% haemolytic rate was ob-
served even after 12 h. In addition, prepared LNPs showing the pro-
duction of H2O2 and interaction with cells in time-bound manner. With
regards to drug loading, pLNPs were able to showed their abilities for
efficient loading of poorly water-soluble drugs and other cytotoxic
agents, e.g. sorafenib and benzazulene (BZL) and at the same time,
increase its release profiles in the pH range of 5.5 and 7.4 with sus-
tainable manner therby enhanced antiproliferation effect. The same
researchers developed carboxylate kraft lignin in order to make the
carboxylated lignin nanoparticles (CLNPs) with a block copolymer
made of PEG, poly(histidine), cell-penetrating peptide and showed its
anticancer effect in tumor cells. On the whole, the LNPs seem to be
promising candidates that can be used to deliver the drug and its su-
perparamagnetic behaviour of Fe3O4-LNPs makes it very powerful and
challenging player in anti-cancer therapy as well as its diagnosis, for
instance, MRI, magnetic targeting etc. (Figueiredo et al. 2017b).
Lievonen et al. (2016) proposed an easy and practical method for
the preparation of pH-sensitive lignin-based complex spherical micelles
in green solvents using pure AL for the encapsulation of Ibuprofen (IBU)
with the help of hydrophobic interaction. The in vitro release behaviour
of IBU was pH-dependent and exhibited controlled release properties. It
is possible to preserve 75% of IBU in simulated gastric fluid whereas in
simulated intestinal fluid, 90% could release smoothly. This work offer
a unique technique for the formulation and fabrication of oral drug
delivery carrier and it possess significant importance in the value-added
use of lignin. Dai et al., 2017showed that AL undergoes self-assembly
with bioactive molecule resveratrol (RSV) and Fe3O4 magnetic nano-
particles, it results in the creation of stable nanodrug carrier. In cyto-
logical as well as animal tests, excellent anti-cancer effects are produced
by the magnetic RSV-loaded lignin nanoparticles (AL/RSV/Fe3O4 NPs)
which further improved the in vitro release and stability of RSV, drug
accumulation, and better tumor reduction (Fig. 5).
Fig. 5. (a) Illustration of self-assembly process of AL/Resveratrol (RSV)/Fe3O4 NPs, (b) SEM and TEM image of AL/RSV/Fe3O4 NPs, and response of AL/RSV/Fe3O4
NPs solution to a magnetic field, (c) RSV release kinetics from different formulations (Dai et al. 2017).
13
Bioresource Technology Reports 9 (2020) 100374
Yiamsawas et al. (2014) generated lignin based polyurethane
hollow nanocapsule at the interface of water–cyclohexane which pos-
sess 2,4-toluene diisocyanate (TDI) and a surfactant. The encapsulating
capacity of the capsules was calculated with the help of hydrophilic
fluorescent dye sulfo-rhodamine and long-time stability was observed
over several months in both aqueous and organic phases. The release of
the dye could be obtained by an enzymatic degradation of the lignin
shell. Interestingly Chen et al. (2018) developed LNP through self-as-
sembly process using renewable and non-toxic aqueous sodium p-to-
luenesulfonate (pTsONa) solution. Number of water-soluble or even
water-insoluble drugs can be dissolved in pTsONa and undergoes 90%
encapsulation and possess sustained drug-releasing capability. In ad-
dition to the above, it is easy to recycle the non-loaded drugs as well as
free pTsONa for multiple times to achieve environmental sustainability.
This synthesis approach with broad processing window could realize
the industrial scale-up production of LNPs and have wide potential
applications.
3.6. As support for enzyme immobilization
Gong et al. (2017) checked the feasibility of a-amylase im-
mobilization supported by lignin of bamboo shoot shells (BSS). It was
interesting to know that BBS work as a α-amylase activator as well as
improve the α-amylase activity twice in the concentration of 5 mg/ml..
Even after reused for 14 times, the residual activity remained compe-
tent enough at 53.2%. Further, Sipponen et al. (2018) prepared cationic
colloidal lignin nanospheres (c-CLP) as activating anchors for hydro-
lases, which is helpful for aqueous ester production through the crea-
tion of spatially-limited biocatalysts in the event of self-assembly and
drying-driven aggregation in calcium alginate hydrogel. The microbial
cutinase and lipase which were spatially-confined had 97% and 70%
synthetic activities when the volume ratio of water to hexane increased
from 1:1 to 9:1 in the reaction medium. Overall above findings work
out for the fabrication of novel, dynamic and renewable biocatalysts to
be used in various applications and can provide insight into segmen-
tation of different heterogenous catalysts.
3.7. As an adsorbent to remove dyes
Azimvand et al. (2018a) prepared lignin nanoparticle-g-polyacrylic
acid adsorbent using copolymerization reactions between lignin nano-
particle and polyacrylic acid in the presence of radical initiator namely
potassium persulfate. The results inferred that the lignin nanoparticle
had adsorption capacity of 99 mg/g whereas it was 138.88 mg/g in case
of lignin nanoparticle-g-polyacrylic acid adsorbent. They further as-
sessed the adsorption capacity of AL and LNPs for the removal of Basic
Red 2 (BR2) from aqueous solutions.. The Langmuir Isotherm was ac-
cepted by both the absorbents AL and LNPs with values such as
P.S. Chauhan
55.2 mg/g and 81.9 mg/g respectively (Azimvand et al. 2018c).
3.8. As a super capacitor
Yu et al. (2018) developed lignin-based microsphere activated and
lignin precursor activated sample (LAC-M and LAC-P) and exploited
their testing it as super capacitors in 6 M KOH solution. The Cyclic
voltammetry (CV) curves of LACs at 50 mV s−1 exhibited approxi-
mately a clean capacitive behaviour of LACs and excellent rate per-
formance as the specific current from 0.05 to 100 A g−1.
3.9. As a nano trap
Yin et al. (2018) create LNPs-gelatin complex by the combination of
Gelatin and LNP for their use as a flocculant. Result shows that complex
shows better flocculation property than LNPs however efficiency of
flocculation is greatly influenced by pH and dosage against Gram +ve
(Staphylococcus aureus) and Gram −ve (Escherichia colii) strains. Ap-
proximately 95% of the flocculation efficiency was attained within
30 min at pH 4.5 whereas it took 1 h for the flocculation efficiency to
reach 90% at pH 5. The LNPs-gelatin complex showed promising results
so that it is a better option to be applied in the flocculation of bacteria
in wastewater treatment, identification of microbes and enrichment of
wastewater.
Recently Xiao et al. (2019) developed lignin-based nano-trap
(LBNT) by lignin functionalization for the removal of various heavy
metal ions. The results showed that there was excellent (> 99%) re-
moval efficiency achieved by LGNT in case of soft Hg(II), Cd(II)) as well
as borderline (Pb(II), Cu(II), Zn(II)) ions.. Furthermore, high amount of
bactericidal rate and antimicrobial rate was showed by LGNT with Ag
(Ag@LBNT) towards E. coli (99.68%) and Staphylococcus aureus
(99.76%). Above work gives clear idea for producing more efficient
cheap biomass-based and application-based nanomaterials as anti-
microbial agents in near future.
4. Conclusion
The field of lignin-based materials has seen significant growth in the
last decade, however, development and usage of lignin for broader
application still faces some challenges, because of its variable chemical
structure and polydispersity. LNPs are one of the lucrative form of the
lignin which shows tremendous opportunity for value added applica-
tions. Unfortunately, most of LNPs preparation methods use hazardous
solvents, therefore it is desirable to replace them with green solvents
namely water and room temperature ionic liquids (RTILs). In near fu-
ture, we need to create eco-friendly methods for LNPs synthesis and
their properties should be optimum for specific applications.
Acknowledgment
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Declaration of competing interest
The author declares that he has no conflict of interest.
References
Agarwal, A., Rana, M., Park, J.M., 2018. Advancement in technologies for the depoly-
merization of lignin. Fuel Process. Technol. 181, 115–132.
Azimvand, J., Didehban, K., Mirshokraie, S.A., 2018a. Safranin-O removal from aqueous
solutions using lignin nanoparticle-g-polyacrylic acid adsorbent: Synthesis, proper-
ties, and application. Adsorpt. Sci. Technol. 36 (7–8), 1422–1440.
Azimvand, J., Didehban, Kh., Mirshokraie, S.A., 2018b. Preparation and characterization
of lignin polymeric nanoparticle using the green solvent ethylene glycol:acid pre-
cipitation technology. BioResources 13 (2), 2887–2897.
14
Bioresource Technology Reports 9 (2020) 100374
Azimvand, J., Didehban, K., Mirshokraie, S.A., 2018c. Preparation and characterization of
nano-lignin biomaterial to remove Basic Red 2 dye from aqueous solutions. Pollution
4 (3), 395–415.
Barakat, A., Putaux, J.L., Saulnier, L., Chabbert, B., Cathala, B., 2007. Characterization of
arabinoxylan-dehydrogenation polymer (synthetic lignin polymer) nanoparticles.
Biomacromolecules 8 (4), 1236–1245.
Barakat, A., Gaillard, C., Lairez, D., Saulnier, L., Chabbert, B., Cathala, B., 2008.
Supramolecular organization of heteroxylan-dehydrogenation polymers (synthetic
lignin) nanoparticles. Biomacromolecules 9 (2), 487–493.
Beisl, S., Miltner, A., Friedi, A., 2017. Lignin from micro- to nanosize: production
methods. Int. J. Mol. Sci. 18, 1244–1274.
Chen, L., Dou, J., Ma, Q., Li, N., Wu, R., Bian, H., Yelle, D.J., Vuorinen, T., Fu, S., Pan, X.,
Zhu, J.Y., 2017. Rapid and near complete dissolution of wood lignin at ≤80°C by a
recyclable acid hydrotrope. Sci. Adv. 3, e1701735.
Chen, L., Zhou, X., Shi, Y., Gao, B., Wu, J., Kirk, T.B., Xu, J., Xue, W., 2018. Green
synthesis of lignin nanoparticle in aqueous hydrotropic solution toward broadening
the window for its processing and application. Chemical. Eng. J. 346, 217–225.
Chen, N., Dempere, L.A., Tong, Z., 2016. Synthesis of pH-responsive lignin-based nano-
capsules for controlled release of hydrophobic molecules. ACS Sustain. Chem. Eng. 4,
5204–5211.
Chung, Y.L., Olsson, J.V., Li, R.J., Frank, C.W., Waymouth, R.M., Billington, S.L., Sattely,
E.S., 2013. A renewable lignin–lactide copolymer and application in biobased com-
posites. ACS Sustain. Chem. Eng. 1, 1231–1238.
Dai, L., Liu, R., Hu, L.Q., Zou, Z.F., Si, C.L., 2017. Lignin nanoparticle as a novel green
carrier for the efficient delivery of resveratrol. ACS Sustain. Chem. Eng. 5,
8241–8249.
Del Saz-Orozco, B., Oliet, M., Alonso, M., Rojo, E., Rodríguez, F., 2012. Formulation
optimization of unreinforced and lignin nanoparticle-reinforced phenolic foams using
an analysis of variance approach. Compos. Sci. Technol. 72, 667–674.
Domenek, S., Louaifi, A., Guinault, A., Baumberger, S., 2013. Potential of lignins as an-
tioxidant additive in active biodegradable packaging materials. J. Polym. Environ.
21, 692–701.
Feldman, D., 2016. Lignin nanocomposites. J. Macromol. Sci. Part. A. 53, 382–387.
Figueiredo, P., Lintinen, K., Kiriazis, A., Hynninen, V., Liu, Z., Bauleth-Ramos, T., et al.,
2017a. In vitro evaluation of biodegradable lignin-based nanoparticles for drug de-
livery and enhanced antiproliferation effect in cancer cells. Biomaterials 21, 97–108.
Figueiredo, P., Ferro, C., Kemell, M., Liu, Z., Kiriazis, A., Lintinen, K., Florindi, H.F., Yli-
Kauhaluoma, J., Hirvonen, J., Kostiainen, A., Santos, H.A., 2017b. Functionalization
of carboxylated lignin nanoparticles for targeted and pH-responsive delivery of an-
ticancer drugs. Nanomedicine. (Lond.) 12 (21), 2581–2596.
Figueiredo, P., Lintinen, K., Hirvonen, J.T., Kostiainen, M.A., Santos, H.A., 2018.
Properties and chemical modifications of lignin: Towards. Lignin-based nanomater-
ials for biomedical applications. Prog. Mater. Sci. 93, 233–269.
Fortunati, E., Yang, W., Luzi, F., Kenny, J., Torre, L., Puglia, D., 2016. Lignocellulosic
nanostructures as reinforcement in extruded and solvent casted polymeric nano-
composites: an overview. Eur. Polym. J. 80, 295–316.
Frangville, C., Rutkevicius, M., Richter, A.P., Velev, O.D., Stoyanov, S.D., Paunov, V.N.,
2012. Fabrication of environmentally biodegradable lignin nanoparticles. Chem.
Phys. Chem. 13, 4235–4243.
Ge, Y., Wei, Q., Li, Z., 2014. Preparation and evaluation of the free radical scavenging
activities of nanoscale lignin biomaterials. BioResources 9, 6699–6706.
Gilca, I.A., Ghitescu, R.E., Puitel, A.C., Popa, V.I., 2014. Preparation of lignin nano-
particles by chemical modification. Iran. Polym. J. 23, 355–363.
Gilca, I.A., Popa, V.I., Crestini, C., 2015. Obtaining lignin nanoparticles by sonication.
Ultrason. Sonochem. 23, 369–375.
Gong, W., Ran, Z., Ye, F., Zhao, G., 2017. Lignin from bamboo shoot shells as an activator
and novel immobilizing support for a-amylase. Food Chem. 228, 455–462.
Gonugunta, P., Vivekanandhan, S., Mohanty, A.K., Misra, M., 2012. A study on synthesis
and characterization of biobased carbon nanoparticles from lignin. World. J. Nano.
Sci. Eng. 2, 148–153.
Gonzalez, M.N.G., Levi, M., Turri, S., Griffini, G., 2017. Lignin nanoparticles by ultra-
sonication and their incorporation in waterborne polymer nanocomposites. J. Appl.
Polym. Sci. 134, 45318.
Gregorová, A., Redik, S., Sedlarík, V., Stelzer, F., 2011. Lignin-containing polyethylene
films with antibacterial activity. In: Proceedings in 3rd International Conference,
Nanocon 2011-Conference, TANGER Ltd. Brno, Czech Republic, EU, Sept 21–23.
Gupta, A.K., Mohanty, S., Nayak, S., 2014. Synthesis, characterization and application of
lignin nanoparticles (LNPs). Mater. Focus. 3, 444–454.
Gupta, A.K., Mohanty, S., Nayak, S.K., 2015. Influence of addition of vapor grown carbon
fibers on mechanical, thermal and biodegradation properties of lignin nanoparticle
filled bio-poly(trimethylene terephthalate) hybrid nanocomposites. RSC Adv. 5,
56028–56036.
Gutiérrez-Hernández, J.M., Escalante, A., Murillo-Vázquez, R.N., Delgado, E., González,
F.J., Toríz, G., 2016. Use of Agave tequilana-lignin and zinc oxide nanoparticles for
skin photoprotection. J. Photochem. Photobiol 163, 156–161.
Hatakeyama, H., Nakayachi, A., Hatakeyama, T., 2005. Thermal and mechanical prop-
erties of polyurethane-based geocomposites derived from lignin and molasses.
Compos. A Appl. Sci. Manuf. 36, 698–704.
Jiang, C., He, H., Yao, X., Yu, P., Zhou, L., Jia, D., 2014. Self-crosslinkable lignin/epox-
idized natural rubber composites. J. Appl. Polym. Sci. 131, 41166.
Juikar, S.J., Vigneshwaran, N., 2017. Extraction of nanolignin from coconut fibers by
controlled microbial hydrolysis. Ind. Crops Prod. 109, 420–425.
Kai, D., Tan, M.J., Chee, P.L., Chu, Y.K., Yap, Y.L., Loh, X.J., 2016. Towards lignin-based
functional materials in a sustainable world. Green Chem. 18, 1175–1200.
Kai, X., Can, J., Gui-feng, L., Gilkes, N., Saddler, J.N., 2015. Preparation and character-
ization of lignin nanoparticles with controllable size by nanoprecipitation method.
P.S. Chauhan
Chem. Ind. For. Prod. 5, 85–92.
Li, H., Deng, Y., Liu, B., Ren, Y., Liang, J., Qian, Y., et al., 2016. Preparation of nano-
capsules via the self-assembly of Kraft lignin: a totally green process with renewable
resources. ACS Sustain. Chem. Eng. 4, 1946–1953.
Li, Y., Qiu, X., Qian, Y., Xiong, W., Yang, D., 2017. pH-responsive lignin-based complex
micelles: Preparation, characterization and application in oral drug delivery.
Chemical. Eng. J. 327, 1176–1183.
Lievonen, M., Valle-Delgado, J.J., Mattinen, M.L., Hult, E.L., Lintinen, K., Kostiainen,
M.A., et al., 2016. A simple process for lignin nanoparticle preparation. Green Chem.
18, 1416–1422.
Liu, H.C., Chien, A.T., Newcom, B.A., Liu, Y., Kumar, S., 2015. Processing, structure, and
properties of lignin- and cnt-incorporated polyacrylonitrile-based carbon fibers. ACS
Sustain. Chem. Eng. 3 (9), 1943–1954.
Liu, Z.H., Hao, N., Shinde, S., Pu, Y., Kang, Y., Kang, X., Ragauskas, A.J., Yuan, J.S., 2019.
Defining lignin nanoparticle properties through tailored lignin reactivity by se-
quential organosolv fragmentation approach (SOFA). Green Chem. 21, 245–260.
Lu, Q., Zhu, M., Zu, Y., Liu, W., Yang, L., Zhang, Y., et al., 2012. Comparative antioxidant
activity of nanoscale lignin prepared by a supercritical antisolvent (SAS) process with
non-nanoscale lignin. Food Chem. 135, 63–67.
Ma, R., Guo, M., Zhang, X., 2018. Recent advances in oxidative valorization of lignin.
Catal. Today 302, 50–60.
Mahmood, N., Yuan, Z., Schmidt, J., Xu, C., 2016. Depolymerization of lignins and their
applications for the preparation of polyols and rigid polyurethane foams: a review.
Renew. Sust. Energ. Rev. 60, 317–329.
Mattinen, M.L., Riviere, G., Henn, A., Nugroho, R.W.N., Leskinen, T., Nivala, O., Valle-
Delgado, J.J., Kostiainen, M.A., Österberg, M., 2018a. Colloidal lignin particles as
adhesives for soft materials. Nanomaterials. (Basel.) 8 (12), 1001.
Mattinen, M.L., Delgado, J.J.V., Leskinen, T., Tuomas, A., Riviere, G., Sipponen, M.,
Paananen, A., Lintinen, K., Kostiainen, M., Osterberg, M., 2018b. Enzymatically and
chemically oxidized lignin nanoparticles for biomaterial applications. Enzym.
Microb. Technol. 111, 48–56.
Melro, E., Alves, L., Antunes, F.E., Medronho, B., 2018. A brief overview on lignin dis-
solution. J. Mol. Liq. 265, 578–584.
Morandim-Giannetti, A.A., Agnelli, J.A.M., Lancas, B.Z., Magnabosco, R., Casarin, S.A.,
Bettini, S.H.P., 2012. Lignin as additive in polypropylene/coir composites thermal,
mechanical and morphological properties. Carbohydr. Polym. 87, 2563–2568.
Myint, A.A., Lee, H.W., Seo, B., Son, W.S., Yoon, J., Yoon, T.J., et al., 2016. One pot
synthesis of environmentally friendly lignin nanoparticles with compressed liquid
carbon dioxide as an antisolvent. Green Chem. 18, 2129–2146.
Nair, S.S., Sharma, S., Pu, Y., Sun, Q., Pan, S., Zhu, J.Y., et al., 2014. High shear homo-
genization of lignin to nanolignin and thermal stability of nanolignin-polyvinyl al-
cohol blends. Chem. Sus. Chem. 7, 3513–3520.
Nypelo, T.E., Carrillo, C.A., Rojas, O.J., 2015. Lignin supracolloids synthesized from (W/
O) microemulsions: use in the interfacial stabilization of Pickering systems and or-
ganic carriers for silver metal. Soft Matter 11, 2046–2054.
Peng, Z., Chen, F., 2011. Synthesis and properties of lignin-based polyurethane hydrogels.
Int. J. Polym. Mater. Polym. Biomater. 60, 674–683.
Pouteau, C., Dole, P., Cathala, B., Averous, L., Boquillon, N., 2003. Antioxidant properties
of lignin in polypropylene. Polym. Degrad. Stab. 81, 9–18.
Pucciariello, R., Bonini, C., D’Auria, M., Villani, V., Giammarino, G., Gorrasi, G., 2008.
Polymer blends of steam-explosion lignin and poly(ε-caprolactone) by high-energy
ball milling. J. Appl. Polym. Sci. 109, 309–313.
Qian, Y., Deng, Y., Qiu, X., Li, H., Yang, D., 2014a. Formation of uniform colloidal spheres
from lignin, a renewable resource recovered from pulping spent liquor. Green Chem.
16, 2156–2163.
Qian, Y., Zhang, Q., Qiu, X., Zhu, S., 2014b. CO2-responsive diethylaminoethyl-modified
lignin nanoparticles and their application as surfactants for CO2/N2-switchable
Pickering emulsions. Green Chem. 16, 4963–4968.
Qian, Y., Qiu, X., Zhong, X., Zhang, D., Deng, Y., Yang, D., et al., 2015a. Lignin reverse
micelles for UV-absorbing and high mechanical performance thermoplastics. Ind.
Eng. Chem. Res. 54, 12025–12030.
Qian, Y., Qiu, X.Q., Zhu, S.P., 2015b. Lignin: a nature-inspired sun blocker for broad-
spectrum sunscreens. Green Chem. 17, 320–324.
Rahman, O., Shi, S., Ding, J., Wang, D., Ahmad, S., Yu, H., 2018. Lignin nanoparticles:
synthesis, characterization and corrosion protection performance. New J. Chem. 42,
3415–3425.
Rangan, A., Manchiganti, M.V., Thilaividankan, R.M., Kestur, S.G., Menon, R., 2017.
Novel method for the preparation of lignin-rich nanoparticles from lignocellulosic
fibers. Ind. Crop. Prod. 103, 152–160.
Richter, A.P., Brown, J.S., Bharti, B., Wang, A., Gangwal, S., Houck, K., et al., 2015. An
environmentally benign antimicrobial nanoparticle based on a silver-infused lignin
core. Nat. Nanotechnol. 10, 817–823.
Richter, A.P., Bharti, B., Armstrong, H.B., Brown, J.S., Plemmons, D., Paunov, V.N.,
Stoyanov, S.D., Velev, O.D., 2016. Synthesis and characterization of biodegradable
lignin nanoparticles with tunable surface properties. Langmuir 32, 6468–6477.
Roopan, S.M., 2017. An overview of natural renewable bio-polymer lignin towards nano
and biotechnological applications. Int. J. Biol. Macromol. 103, 508–514.
Sameni, J.K., 2015. Physico-Chemical Characterization of Lignin Isolated from Industrial
Sources for Advanced Applications. PhD thesis. Faculty of Forestry, University of
Toronto.
Schutyser, W., Renders, T., Van den Bosch, S., Koelewijn, S.F., Beckham, G.T., Sels, B.F.,
2018. Chemicals from lignin: an interplay of lignocellulose fractionation, depoly-
merisation, and upgrading. Chem. Soc. Rev. 47, 852–908.
Silmore, K.S., Gupta, C., Washburn, N.R., 2016. Tunable Pickering emulsions with
polymer-grafted lignin nanoparticles (PGLNs). J. Colloid Interface Sci. 466, 91–100.
15
Bioresource Technology Reports 9 (2020) 100374
Sipponen, M.K., Smyth, M., Leskinen, T., Johansson, L.S., Österberg, M., 2017. All-lignin
approach to prepare cationic colloidal lignin particles: stabilization of durable
Pickering emulsions. Green Chem. 19, 5831–5840.
Sipponen, M.K., Farooq, M., Koivisto, J., Pellis, A., Seitsonen, J., Österberg, M., 2018.
Spatially confined lignin nanospheres for biocatalytic ester synthesis in aqueous
media. Nature. Comm. 9, 2300.
Sun, Y., Yang, L., Lu, X., He, C., 2015. Biodegradable and renewable poly (lactide)–lignin
composites: synthesis, interface and toughening mechanism. J. Mater. Chem. A 3,
3699–3709.
Tian, D., Hu, J., Bao, J., Chandra, R.P., Saddler, J.N., Lu, C., 2017a. Lignin valorization:
lignin nanoparticles as high-value bio-additive for multifunctional nanocomposites.
Biotechnol. Biofuels. 10, 192.
Tian, D., Hu, J., Chandra, R.P., Saddler, J.N., Lu, C., 2017b. Valorizing recalcitrant cel-
lulolytic enzyme lignin via lignin nanoparticles fabrication in an integrated bior-
efinery. ACS Sustain. Chem. Eng. 5, 2702–2710.
Tortora, M., Cavalieri, F., Mosesso, P., Ciaffardini, F., Melone, F., Crestini, C., 2014.
Ultrasound driven assembly of lignin into microcapsules for storage and delivery of
hydrophobic molecules. Biomacromolecules 15, 1634–1643.
Upton, B.M., Kasko, A.M., 2016. Strategies for the conversion of lignin to high-value
polymeric materials: review and perspective. Chem. Rev. 116, 2275–2306.
Wang, B., Sun, D., Wang, H.M., Yuan, T.Q., Sun, R.C., 2019b. Green and facile prepara-
tion of regular lignin nanoparticles with high yield and their natural broad-spectrum
sunscreens. ACS Sustain. Chem. Eng. 7, 2658–2666.
Wang, H., Pu, Y., Ragauskas, A., Yang, B., 2019a. From lignin to valuable products–-
strategies, challenges, and prospects. Bioresour. Technol. 271, 449–461.
Xiao, D., Ding, W., Zhang, J., Ge, Y., Wu, Z., Li, Z., 2019. Fabrication of a versatile lignin-
based nano-trap for heavy metal ion capture and bacterial inhibition. Chemical. Eng.
J. 358, 310–320.
Xing, Q., Buono, P., Ruch, D., Dubois, P., Wu, L., Wang, W.J., 2019. Biodegradable UV-
blocking films through core−shell lignin− melanin nanoparticles in poly(butylene
adipate-co-terephthalate). ACS Sustain. Chem. Eng. 7, 4147–4157.
Yang, M., Zhao, W., Singh, S., Simmons, B., Cheng, G., 2019. On the solution structure of
Kraft lignin in ethylene glycol and its implication for nanoparticle preparation.
Nanoscale. Adv. 1, 299–304.
Yang, W., Kenny, J.M., Puglia, D., 2015a. Structure and properties of biodegradable
wheat gluten bionanocomposites containing lignin nanoparticles. Ind. Crop. Prod. 74,
348–356.
Yang, W., Fortunati, E., Dominici, F., Kenny, J.M., Puglia, D., 2015b. Effect of processing
conditions and lignin content on thermal, mechanical and degradative behavior of
lignin nanoparticles/polylactic (acid) bionanocomposites prepared by melt extrusion
and solvent casting. Eur. Polym. J. 71, 126–139.
Yang, W., Dominici, F., Fortunati, E., Kenny, J.M., Puglia, D., 2015c. Effect of lignin
nanoparticles and masterbatch procedures on the finalproperties of glycidyl metha-
crylate-g-poly (lactic acid) films beforeand after accelerated UV weathering. Ind.
Crop. Prod. 77, 833–844.
Yang, W., Fortunati, E., Dominici, F., Giovanale, G., Mazzaglia, A., Balestra, G.M., Kenny,
J.M., Puglia, D., 2016a. Synergic effect of cellulose and lignin nanostructures in PLA
based systems for food antibacterial packaging. Eur. Polym. J. 79, 1–12.
Yang, W., Owczarek, J.S., Fortunati, E., Kozanecki, M., Mazzaglia, A., Balestra, G.M.,
Kenny, J.M., Torre, L., Puglia, D., 2016b. Antioxidant and antibacterial lignin na-
noparticles in polyvinylalcohol/chitosan films for active packaging. Ind. Crop. Prod.
94, 800–811.
Yang, W., Rallini, M., Wang, D.Y., Gao, D., Dominici, F., Torre, L., Kenny, J.M., Puglia, D.,
2018a. Role of lignin nanoparticles in UV resistance, thermal and mechanical per-
formance of PMMA nanocomposites prepared by a combined free-radical graft
polymerization/masterbatch procedure. Compos. Part A 107, 61–69.
Yang, W., Fortunatia, E., Bertoglio, F., Owczarek, J.S., Brunif, G., Kozanecki, M., Kenny,
J.M., Torre, L., Visai, L., Puglia, D., 2018b. Polyvinyl alcohol/chitosan hydrogels with
enhanced antioxidant and antibacterial properties induced by lignin nanoparticles.
Carbohydr. Polym. 181, 275–284.
Yang, Y., Deng, Y., Tong, Z., Wang, C., 2014. Renewable lignin based xerogels with self
cleaning properties and superhydrophobicity. ACS Sustain. Chem. Eng. 2,
1729–1733.
Yearla, S.R., Padmasree, K., 2016. Preparation and characterisation of lignin nano-
particles: evaluation of their potential as antioxidants and UV protectants. J. Exp.
Nanosci. 11, 289–302.
Yiamsawas, D., Baier, G., Thines, E., Landfester, K., Wurm, F.R., 2014. Biodegradable
lignin nanocontainers. RSC Adv. 4, 11661–11663.
Yin, H., Liu, L., Wang, X., Wang, T., Zhou, Y., Liu, B., Shan, Y., Wang, L., Lü, X., 2018. A
novel flocculant prepared by lignin nanoparticles-gelatin complex from switchgrass
for the capture of Staphylococcus aureus and Escherichia coli. Colloids Surf. A
Physicochem. Eng. Asp. 545, 51–59.
Yu, B., Chang, Z., Zhang, Y., Wang, C., 2018. Preparation and formation mechanism of
size-controlled lignin based microsphere by reverse phase polymerization. Mater.
Chem. Phys. 203, 97–105.
Yu, J., Wang, J.F., Wang, C.P., Liu, Y.P., Xu, Y.Z., Tang, C.B., Chu, F.X., 2015. UV-ab-
sorbent lignin-based multi-arm star thermoplastic elastomers. Macromol. Rapid
Commun. 36, 398–404.
Zhao, W., Simmons, B., Singh, S., Ragauskas, A., Cheng, G., 2016. From lignin association
to nano−/micro-particle preparation: extracting higher value of lignin. Green Chem.
18, 5693–5700.
Zikeli, F., Vinciguerra, V., D’Annibale, A., Capitani, D., Romagnoli, M., Mugnozza, G.S.,
2019. Preparation of lignin nanoparticles from wood waste for wood surface treat-
ment. Nanomaterials 9, 281.