Recent Patents on Food, Nutrition & Agriculture, 2011, 3, 1-8 1

Recent Patents on Eggshell: Shell and Membrane Applications

Cristianne M.M. Cordeiro and Maxwell.T. Hincke*

Department of Cellular and Molecular Medicine, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5, Canada

Received: August 18, 2010; Accepted: September 9, 2010; Revised: October 16, 2010
Abstract: The chicken eggshell and its membranes are an inexpensive and abundant waste material which exhibit
interesting characteristics for many potential applications. The eggshell is formed mainly of calcium carbonate (CaCO3)
and is used widely as an animal feed, lime (Ca(OH)2) substitute or a fertilizer. Moreover, the associated eggshell
membranes have a high content of bioactive components, as well as properties of moisture retention and biodegradability
which have potential use for clinical, comestic, nutraceutical and nanotechnology applications. The eggshell membranes
have been also used for biosorption of heavy metals and dyes and as a template to synthesize metal nanoparticles. The
combination of nanosized calcium phosphate (Ca3(PO4)2) biomaterials synthesized from eggshell and eggshell membrane
show promise to develop drug delivery system and nanowires for electronic devices. In addition, a derived product, the
soluble eggshell membrane protein (SEP) has applications in tissue engineering. This review discusses the patented
applications of eggshell membrane waste: shell and membrane for the last 10 years as well as their future applications.
Keywords: Bioremediation, clinical, collagen, cosmetics, eggshell membrane, nanotechnology, nutraceutical and shell.

INTRODUCTION plant [10]. The eggshell residue generated by these opera-

tions was historically useless and commonly disposed of in
The avian eggshell structure is composed of shell and
landfill without any pre-treatment. However, this waste
membrane and represents about 10% of the egg weight. The
management is not a desirable practice in view of the
shell is a calcareous structure predominantly constituted of
environmental odour from biodegradation. Egg-based
calcium carbonate(CaCO3) (95%) and an organic matrix processing companies have been investing in research and
composed of proteins, glycoprotein and proteoglycans
development of processes to extract products with potential
(3.5%) [1, 2]. The shell is composed of calcitic columns
commercial application from their waste. The eggshell
which are organized in two parts: a major part with roughly
membrane waste is a source of active compounds which are
parallel sides (the palisade layer) and a minor part forming
naturally concentrated and can be extracted using eco-
the rounded inner ends (mammillary cone layer) that are
friendly and cost effective processes. A number of processes
attached to the fibres of the shell membranes (Fig. (1A). The for harvesting the application potential of eggshell mem-
outermost layer of the chicken eggshell is the cuticle, formed
brane waste have been patented. Table 1 [11, 18], Table 2
by vesicles that are >85% proteinaceous [3]. The eggshell
[19, 32] and Table 3 [33, 44] show relevant patent appli-
membranes are found between the egg white and the
cations for shell and membranes that are further discussed in
calcified shell, and are thought to play an important role in
the next sections of this article.
determining the origin and ultrastructure of the calcified
shell [4, 5]. The collagen fibres of the eggshell membrane
are organized into two individual membranes: a thin inner I. EGGSHELL & PROCESSING
one located upon the egg white (albumen), and a thick outer Egg shell waste has been used as animal feed, lime
membrane attached to the shell Fig. (1B-D). The eggshell substitute or a fertilizer. Eggshell waste is a promising
membrane consists of cross-linked collagens (I, V and X), fertilizer because only a small amount of eggshell can have a
glycosaminoglycans (GAGs), egg white proteins (i.e. positive impact on the fertilization process [19]. In many
Ovotransferrin, Lysozyme) and eggshell matrix proteins (i.e. countries, it is accepted practice for egg shells to be dried at
Ovocalyxin-36) [1, 4, 6-8]. Hydroxyproline, hydroxylysine high temperature to be decontaminated for use as a source of
and desmosine are detected in its acid hydrolysates [9]. calcium in animal feeds. A polypropylene composite with
In the egg producing industry, a large percentage of eggs crushed eggshell as filler has been recently fabricated and
are diverted to breaker operations to produce liquid egg characterized for industrial applications [20, 45]. Shells have
products and are not consumed as shell eggs (i.e. 32% in been dried, crushed, acid-treated, abraded and tumbled, but
United States, close to 25% in Europe). The by-products of the membrane remains tenaciously attached to the shell by
these operations are the eggshell and associated membranes, the outer membrane fibres that penetrate the mammillary
which constitute approximately 6g / egg; this represents a cones. However, considerable progress has been made in
significant waste from the egg-derived products processor eggshell processing and some promising eggshell processing
methods have been patented. A simple and economic method
*Address correspondence to this author at the Department of Cellular and based on differential flotation of shell fragments and mem-
Molecular Medicine, University of Ottawa, 451 Smyth Road, Ottawa, K1H brane particles in water was proposed by MacNeil [11]
8M5, Canada; Tel: +1 613 562 5800, x8193; Fax: +1 613 562 5687;
E-mail: mhincke@uottawa.ca

1876-1429/11 $100.00+.00 © 2011 Bentham Science Publishers Ltd.

2 Recent Patents on Food, Nutrition & Agriculture, 2011, Vol. 3, No. 1 Cordeiro and Hincke

Fig. (1). Scanning electron micrographs illustrating the morphology of the eggshell and eggshell membranes. (A) Eggshell cross-fractured to
reveal the shell membrane (SM), mammillary layer (ML) and palisade layer (PL); (B) higher magnification of the membrane/mammillary
body interface. Outer shell membrane fibres (OSM) insert into the tips of the mammillary bodies (MB). Inner shell membranes (ISM); (C)
Enlargement of the shell membrane fibres (SMF), revealing their interwoven and coalescing nature; (D) inner aspect of the inner shell
membrane (ISM), demonstrating the limiting membrane (LM) that surrounds the egg white (here removed) during sample preparation. Scale
bars: a, 50 μm; b, 20 μm; C and D, 2 μm.
Reprinted from Matrix Biology,19, Hincke MT, Gautron J, Panheleux M, Garcia-Ruiz J, McKee MD, Nys Y, Identification and localization
of lysozyme as a component of eggshell membranes and eggshell matrix, 443-453, Copyright (2000), with permission from Elsevier.
Table 1. Relevant Patents for Shell and Membranes.

Shell and Membranes Patent Reference No.

Separation of membranes from shell based on differential flotation [11]

Separation of membranes and shell from hatchery eggshell waste by particle size reducing [12]

Separation of membranes and shell with processing chamber impact zone [13]

Separation of membranes and shell with pulsed energy separation unit [14]

Separation of membranes and shell with cavitation [15]

Separation of membranes and shell with overlapping vanes [16]

Separation of membranes and shell using solutions of EDTA and hexane [17]

Pulverized eggshell membrane and chitosan as bio-cream and lotion [18]

Applications for Eggshell & Membranes Recent Patents on Food, Nutrition & Agriculture, 2011, Vol. 3, No. 1 3

(Table 2) [33, 44] who patented a device for abrading the useful amounts of a large number of microelements such as
membrane from shell and a tank with water where the strontium (Sr), fluorine (F) and selenium (Se) [48]. The
eggshell and membranes are passively separated. The health benefits of consuming chicken eggshell powder as a
eggshell particles tend to settle to the bottom of the tank and calcium supplement have shown significant and beneficial
the membrane remains in the liquid. After that, two devices results for decreasing bone loss. Clinical studies with post-
communicate with the tank: one for shell recovery and other menopausal women and in persons with osteoporosis or
for membranes recovery. Some optimization for eggshell osteopenia showed that the intake of calcium extracted from
membrane separation was developed using a vacuum suction chicken eggshell powder is associated with increases in bone
system to recover hatchery waste shells from live chicks and mineral density (BMD) of hips and lumbar spine [48, 49].
unhatched eggs [12] or even using a processing chamber to Eggshell powder has also been compared to purified calcium
create impact zone conditions where the size of remaining carbonate as a calcium source in animal feed for piglets [50].
eggshell is reduced [13]. Moreover, the separation of shell A heating process was developed to obtain eggshell powder
from membranes in the liquid medium was also improved from waste eggshell in order to use as a calcium supplement
with the application of a pulsed energy separation system in food and drink [22] Table 2. A patented process claims the
[14] or the application of cavitation [15]. The use of multiple use of food grade hydrochloric acid to extract calcium
overlap-ping vanes of defined specific gravity in the interior carbonate from eggshell providing calcium carbonate
of tanks where the membranes are separated from shell product without heavy metals in its composition for clinical
mineral allows total recovery of membrane and shell [16]. A and nutraceutical applications [23]. The eggshell powder
chemical method to separate the membrane from shell waste is also a source for the synthesis of new compounds
involving the use of a combination of EDTA and hexane has such as calcium glutamate which can be used as calcium
been patented [17]. Separation of eggshell into its mineral supplement, salt substitute and flavoring agent [24].
and membrane constituents allows different applications for Monocalcium and dicalcium phosphate were produced using
each material to be developed. For example, the membrane- eggshell and can be used as calcium and phosphate
free shell powder can be used in the paper industry as supplements for farm animal feed [25]. New organic calcium
coating pigments for ink-jet printing paper [46], as chalk for with good absorption by the human body such as calcium
drawing on blackboard which does not chap the skin fumarate and calcium maleate were obtained using eggshell
compared to the common chalk [21] or in agriculture as lime [26]. Eggshell as a bone substitute suitable for grafting to
substitute [47]. promote bone healing has been explored in animal models
using rat calvaria and cranial defects in rabbits [51, 52]. A
II. NUTRACEUTICAL & CLINICAL APPLICATIONS chemical extraction of active polypeptides from soft and
hard eggshell tissue was patented [27] for application to
Shell stimulate osteoblast growth. These eggshell polypeptides
Eggshell is a great source of dietary calcium; it is showed potential benefit on osteoblast cell growth which
composed of about 39% elemental Ca which also contains suggests a possible oral medication for treatment of bone

Table 2. Relevant Patents for Shell.

Shell Patent Reference No.

Shell as fertilizer [19]

Shell powder as filler for polypropylene composites [20]

Shell powder as chalk [21]

Shell powder as calcium supplement [22]

Synthesis of calcium carbonate [23]

Synthesis of calcium glutamate [24]

Synthesis of monocalcium and dicalcium phosphate [25]

Synthesis of organic calcium [26]

Extraction of eggshell polypeptides to osteoblasts cell growth [27]

Removal of heavy metals from water [28]

Removal of heavy metals from soil using pulverized shell [29]

Eggshell microparticles as facial cleanser [30]

Eggshell as friction agent [31]

Synthesis of ceramic powder containing hydroxyapatite nanoparticles from calcined eggshell [32]
4 Recent Patents on Food, Nutrition & Agriculture, 2011, Vol. 3, No. 1 Cordeiro and Hincke

disorders. This promising result leads to other possible III. BIOREMEDIATION

applications for these polypeptides including growing bone
The limitations and high investments (i.e. installations
cells ex vivo and bone grafting in surgery.
and operating cost) necessary for some traditional techniques
to remove dyes and heavy metals from contaminated
Membranes aqueous environments (chemical precipitation, coagulation,
Natural eggshell membrane (NEM) has been used as a ion exchange, etc.) have drawn attention to nonconventional
dietary supplement for nutraceutical treatment. Ingestion of alternative techniques [58]. The application of natural waste
glycosaminoglycans and proteins from chicken eggshell products to solve this environmental problem is an especially
membrane are suggested to be beneficial for healthy joints promising strategy. A number of studies have demonstrated
and regulation of connective tissues. Clinical trials that eggshell membranes bind metal ions and other positively
demonstrate that patients with knee osteoarthritis receiving charged molecules (perhaps due to immobilized ovotrans-
500mg of NEM daily experienced a noticeable reduction in ferrin content). Ground eggshell waste may be utilized as
joint pain as well as stiffness, compared to the placebo [53, low cost adsorbent for the removal of anionic dyes or heavy
54]. Eggshell membrane has been evaluated as a biode- metals from aqueous solution [59, 60].
gradable bone regeneration inhibitor [55]. Glycosamino- For successful use of eggshell powder as a biological
glycans present in the eggshell membrane such as hyaluronic sorbent, it is important to characterize features such as
acid have been isolated due to the promising application for particle size, shape and porosity [61, 62]. Finely powdered
treating diseases such as osteoarthritis and periodontal eggshell was more effective than coarse particles to adsorb
diseases [33, 34] Table 3. Other patent claims report a anionic dyes from water samples (BET surface area of 21
method that evaluates the inhibitory effect of processed and 1.023m2/g, respectively). Eggshell was also tested for
eggshell membrane and eggshell membrane isolates (i.e. sorption of heavy metals such as Fe (III) ions from aqueous
collagen, glucosamine, chondroitin sulfate and hyaluronic solutions. A patented process for treating wastewater (heavy
acid) on the expression of proinflammatory cytokines in metals) using ground eggshell showed a heavy metal
mammals. It was observed that these substances isolated removal efficiency of 50% - 90%, where the process does
from eggshell membrane showed anti-inflammatory pro- not require special working conditions [28]. Soils conta-
perties based on reduction of pro-inflammatory plasma minated with heavy metals can be also remediated by the use
cytokine levels [35, 36]. Some compounds extracted from of pulverized eggshell. A patent reports that mixing
the hydrolysis of eggshell membrane by proteases may be pulverized eggshell with soil contaminated with heavy metal
used for treating diseases such as hypertension [37]. In some can reduce the heavy metal concentration in the soil as well
cases, the eggshell membrane powder extracted using a as improve the quality of soil [29]. Some components
simple method which maintains the original nutritional extracted from natural eggshells have been also considered
compounds was used for treatment of gastrointestinal as promising absorbents for heavy metals removal. The
disorders [38]. It is reported that fresh eggshell membrane potential applicability of calcined eggshell (99.63% lime) to
can be utilized as an antimicrobial tissue adhesive to aid remove heavy metals from synthetic and natural wastewater
healing of certain skin lacerations [39, 56, 57]. Fibers made was reported by Park and coworkers [58]. Both natural
from eggshell membrane have been developed by an eggshell and calcined eggshell showed impressive capacity
electrospinning method. The fibers strongly adhere to skin to remove heavy metals. However, calcined eggshell showed
tissue, indicating potential use as wound dressing [40]. higher efficiency even in drastic conditions such as strong

Table 3. Relevant Patents for Membranes.

Membranes Patent Reference No.

Extraction of hyaluronic acid from membranes [33]

Treatment of osteoarthritis with hyaluronic acid isolates from membranes [34]

Extraction of bioactive components from membranes with anti- inflammatory properties [35,36]

Treatment of diseases such as hypertension with hydrolysed membrane [37]

Membrane component powder to treat gastrointestinal disorders [38]

Hydrolysed membranes as adhesive plaster to treat wounds [39]

Membranes as fiber to produce wound dressing [40]

Membranes as template for carbon doped TiO2 porous nanometer photocatalyst [41]

Hydrolysed membrane as cream or powder makeup [42]

Hydrolysed membrane and collagen as cosmetic to prevent wrinkle formation [43]

Therapeutic, nutraceutical and cosmetics applications for eggshell membrane and processed eggshell membrane [44]
Applications for Eggshell & Membranes Recent Patents on Food, Nutrition & Agriculture, 2011, Vol. 3, No. 1 5

acid wastewater. Carbonate hydroxylapatite (CHAP) and chitosan [18]. Moreover, eggshell has also been patented
(Ca5(PO4CO3)3OH) synthesized using eggshell waste also as cosmetic products such as a facial cleansing agent [30]
shows high affinity for heavy metals including cadmium, and friction agent [31]. Eggshell particles showed better effi-
copper and lead ions present in aqueous solution [63, 64]. ciency to remove sebum and dust from clogged skin pores in
The carbon doped TiO2 porous nanometer photocatalyst can comparison to other facial cleansers.
be successfully applied to degrade organic contaminants in
industrial sewage [41]. VI. NANOTECHNOLOGY: NEW BIOMATERIALS,
COMPOSITES AND DRUG DELIVERY
IV. CHEMICAL PROCESSING SUPPORT
Shell and eggshell membranes have been a starting point
Eggshell membranes can be used as natural solid phase for synthesis of new biomaterials. The synthesis of bioactive
support for biomimetic chemical processing in a variety of materials based on calcium phosphate (CaP) structures using
nanotechnology applications. Many studies have shown the eggshell as the main source of calcium have been employed
use of eggshell membrane to synthesize metal nanoparticles in many clinical applications [70-72]. Among many CaP
(NPs) of various core sizes which have many applications structures, hydroxyapatite (Ca10(PO4)6(OH)2 is well known
for catalysis, medical imaging and optics [65-68]. For as a biocompatible material with chemical similarity to the
instance; barium nano-structures of chromate (BaCrO4) may mineral component of bones and teeth. The hydroxyapatite
be synthesized using eggshell membrane as a biotemplate produced from eggshell has been used as bioinert implants,
[65]. In situ synthesis of lead sulphide (PbS) nanoclusters for prosthetics and bone filler [72, 73]. Moreover, when hydro-
optoelectronics applications has been achieved on eggshell xyapatite is synthesized as nanostructures, the biocom-
membrane fibres [66]. In a study, thin films with a macro- patibility of hydroxyapatite is enhanced with a consequent
porous network structure made up of crystalline ZrO2 were increase in the range of applications [74]. For instance,
synthesized using eggshell membrane template. ZrO2 was eggshell-derived hydroxyapatite nanoparticles have been
coated onto eggshell membranes by the sol-gel minerali- used as a scaffold for tissue engineering, suitable drug
zation method. The assembly of biotemplate and sol-gel delivery agent and as carriers for non-viral gene delivery
approach is very effective since the synthetic metal oxides [74-76]. The synthesis of hydroxyapatite nanopowders from
with similar structure to natural metal oxides can be eggshell and also mollusk shells using mechanochemical
produced [67, 68]. In another application, electrodialysis of activation was developed [77]. Nano-hydroxyapatite can also
racemic mixtures of amino acids through eggshell mem- be produced from calcined eggshell by a hydrothermal
branes resulted in selective chiral separation [69]. A patent method with a cationic surfactant template [74]. The produc-
claims the use of eggshell membrane as template for pre- tion of ceramic powder containing nano-hydroxyapatite
paring a carbon doped TiO2 porous nanometer photocatalyst particles from calcined eggshell for use as a bone substitute
[41]. is the subject of a patent claim [32]. Nanocrystalline
hydroxyapatite synthesized from eggshell waste showed
V. COSMETICS improvements in some properties such as stability at high
temperatures and osteoblast cell adhesion compared to syn-
Eggshell membranes are a potential source of bioactive thetic and commercial hydroxyapatite [78]. Eggshell mem-
proteins, proteoglycans and glycosaminoglycans which are brane has been employed as a biotemplate for the develop-
associated with the fibers composed of collagen types I, V ment of metallic nanostructures such as a Tris (8-
and X. These compounds are present in eggshell membrane hydroxyquinoline) aluminum (III) (Alq3) nanowires [79].
in significant amounts compared to other known sources and Alq3 nanowires can be used as building blocks for electronic
may require less processing to isolate them [4, 44]. Collagen devices [80]. A combination of hydroxyapatite and
extracted from eggshell membrane has potential applications nanoribbon spherulites treated with fluorescein using
in food, cosmetic and biomedical industries due to its tested eggshell membrane template might be a promising candidate
safety properties including low autoimmune and allergic for a delivery system to be used for medical and pharma-
reactions compared to collagen extracted from other sources ceutical uses [81]. Hydroxyapatite nanoribbon spherites were
[33]. Many cosmetics are based on eggshell membrane and also used to produce nanocomposite spheres containing
collagens which are hydrolysed to generate an eggshell silver nanoparticles using eggshell membrane as template.
membrane hydrolysate. Such a hydrolysate is composed of These nanocomposite spheres formed with hydroxyapatite
hexosamine, chondroitin sulfate, hyaluronic acid, collagen, improve the antibacterial effect of silver nanoparticles [82].
and/or other proteins and has been the main composition of
some cosmetics for example: emulsion, cold cream or
VII. CURRENT& FUTURE DEVELOPMENTS
powder makeup [44]. In other patents, eggshell membrane
hydrolysate was used to produce cosmetics possessing great The eggshell membrane is a promising natural bioma-
promise to prevent the formation of skin wrinkles [43] as terial which can be used for nutraceutical, clinical,
well as improvement of the elasticity and thickness of skin bioremediation, chemical process support and cosmetic
[42]. Combining various active compounds is becoming a applications. Despite its many virtues, the shape, size and
trend to enhance the efficacy of cosmetic products and has thickness of eggshell membranes cannot be modified, and
become more attractive for large scale commercialization. insolubility due to the presence of disulfide bounds in its
For instance, a patent claims that a water soluble bio-cream / molecular structure considerably reduces the range of
bio-lotion which activates skin can be produced by com- eggshell membrane applications [83]. However, many
bining lactic acid - treated pulverized eggshell membrane studies have considered a soluble eggshell membrane protein
6 Recent Patents on Food, Nutrition & Agriculture, 2011, Vol. 3, No. 1 Cordeiro and Hincke

(SEP) as an alternative biomaterial [84, 85].The preparation [14] Oren, E. Eggshell membranes and eggshells separation system
of SEP is based on the dissolution of eggshell membrane by comprises supply conduit, pulsed energy separation unit, and
separator for separating membranes from liquid medium.
reductive cleavage of disulfide bonds [83] and causes US126912 (2010).
modification of the morphology, thickness and solubility of [15] Vlad, V. Eggshell membrane separation method. US7584909
the eggshell membrane. SEP has been extensively used in (2009).
tissue engineering and in the biomedical field [83, 86]. [16] Floh, R.V., Jalfen, S.L. Apparatus for separating the organic
membrane portion and the mineral portion of broken egg shells.
Another future potential application for eggshell membranes US206009 (2009).
is in the pharmaceutical domain, since Ovocalyxin-36 [17] Nakkarike, M.S., Narayan, B., Patiram, Z.S., Namadev, S.M.
(OCX-36), a novel potential antibacterial protein associated Process for separation of shell membrane from wet egg shells.
with the eggshell membrane was recently identified. IN00315 (2006).
Analysis of its protein sequence showed significant identity [18] Yozo, I. Moisturising lotion or cream contg. aminoacid - is prepd.
by adding chitosan to soln. obtd. by diluting eggshell with water or
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understanding of the properties of the purified protein are pylene composites with reinforcement based on eggshells:
required to define potential therapeutic applications of procedure to obtain the said composite, reinforcement based on
OCX-36. eggshells, and procedure for obtaining it. US7459492 (2008).
[21] Simo, H. Chalk for drawing picture on blackboard, is obtained by
removing egg membrane from eggshell, drying, grinding, mixing
ACKNOWLEDGMENTS sizing agent with ground product, and solidifying. JP120280
The authors would like to thank the support provided by (2005).
[22] Kimura, Y. Eggshell powder useful as calcium supplement in pet
the Canadian Natural Sciences and Engineering Research foods, tablet, foodstuff and drink, obtained by heat-processing shell
Council, Ontario Ministry of Agriculture, Food and Rural of hen's egg, removing thin layer and obtaining fine powder of the
Affairs, and the Poultry Industry Council. eggshell. JP017827 (2008).
[23] Yu, J. Biological calcium carbonate extract for medical, health-
care, and food additives, comprises eggshell or shell, and edible
CONFLICT OF INTEREST hydrochloric acid. CN101554387 (2009).
[24] Wang, J., Wei, Y. Synthesis of calcium glutamate chelate for use as
The authors declare no conflict of interest. calcium supplement, salt substitute and flavoring agent comprises
using shell processing waste as calcium source, reacting glutamic
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