Title: Non-rubbers in Natural Rubber Latex

Author: Eng Aik Hwee

Affiliate: Ansell Shah Alam, Malaysia,

Lot 16, Persiaran Perushaan, Section 23, Shah Alam, 40000, Selangor,
Malaysia

e-mail: engah@ap.ansell.com
Introduction
The major components of fresh natural rubber latex are water and cis 1,4-
polyisoprene. The rubber hydrocarbon content, i.e., dry rubber content (DRC),
normally in the range of 30-40% is determined by coagulating the latex with an
organic acid, such as acetic or formic acids followed by washing, and drying in an
oven. The total solid content (TSC), however, is determined by drying the latex in the
oven, removing the volatile materials. It is normally about 5% higher than DRC due
to the presence of non-rubbers in the latex. There are many factors that can affect the
non rubbers content of natural rubber and these include clone, soil and climate
conditions, wintering season, type of fertilizers used, and tapping frequency. A typical
composition of natural rubber latex is given in Table 1.

Table 1: Composition of fresh latex

Component Percentage, w/w
Rubber hydrocarbon 36.0
Proteins, amino acids & nitrogenous compounds 1.7
Lipids 1.6
Ash 0.5
Inositols & carbohydrates 1.6
Water 58.6

Furthermore, the non-rubbers can also be chemically bonded to the main-chain

rubber molecule. They are known as the abnormal groups of natural rubber, i.e. non
cis isoprene group. According to the known biosynthesis pathway for the naturally
occurring cis polyisoprene, the initiating group, or the -terminal is expected to be a
dimethylallyl while the terminating group, or -terminal is an alcohol, or fatty ester
groups.
Although the roles of these non-rubbers and abnormal groups in the latex are
yet to be fully understood, they are believed to be of biological significance with
regard to the structure and biosynthesis pathway of the rubber. In addition, they also
known to affect the physical and chemical properties of the latex, dry rubber and
rubber products. This makes natural rubber a unique material that is difficult to be
replicated synthetically. Since the review on the abnormal groups has been presented
recently,1 this paper focuses on the composition of various non-rubbers and their
effects on natural rubber properties.

Lipids
Natural rubber lipids, which comprise of neutral and polar lipids, make up the
largest proportion of non-rubbers in the latex. Depending on the clonal origin of the
latex, the amount of lipids isolated from different latices has been found to vary from
1.3 – 3.5%w/w. The lipids that are adsorbed on the surface of rubber particles are
normally found in the cream fraction of centrifuged latex. In contrast, the rubber
particles in the skim latex, which are of smaller size, have a very low level of
adsorbed lipids.2-3 Drying natural rubber by smoking, has been found to increase the
amount of extractable lipid but reduce the free fatty acid content.4

Neutral lipids
Triglycerides are the largest component of the neutral lipids. The others are
esters, phenolic compounds (tocotrienols), sterols. In addition, the neutral lipids of
natural rubber have also been found to contain free -tocotrienol and -tocotrienol,
and esterified -tocotrienol and -tocotrienol.5 The total amount of tocotrienols in the
dry natural rubbers varies from 0.02% to 0.15%, depending on the clonal origin of the
latex. Tocotrienols together with other phenolic compounds from the unsaponifiable
fraction are the most important natural antioxidant, preventing the autoxidation of the
raw rubber.6 The minor components of the neutral lipids are free fatty acids, fatty
alcohols, monoglycerides, diglycerides, and carotenoid pigments. The free fatty acids
are mainly stearic, oleic and linoleic acids.7-10 The free fatty acids are believed to be
the breakdown products of lipids. They could form soaps with cations and adsorbed
onto the rubber particles, contributing to the colloidal stability of the latex. On the
other hand, the free fatty acids and unsaturated methyl fatty esters have also been
reported to accelerate the oxidation and chain scission of deproteinized natural
rubber.11 Certain free fatty acids, such as stearic and linolenic, are activators of
sulphur vulcanization. These compounds give better heat build-up properties to
natural rubber, 12 and also function as crystallization nuclei,13-17 plasticizer,18 and
softener.19 However, when added to natural rubber, the long chain fatty acids do not
contribute to the stretch induced crystallization of the rubber, a property that enhances
the physical strength of natural rubber.20

Glycolipids
The glycolipids in natural rubber vary from 0.3 – 1.0%, depending on the
rubber clones. The four main components of glycolipids are digalactosyl diglyceride,
steryl glucosides, esterified steryl glucosides, and monogalactosyl diacylglycerols.
The sterols are mainly stigmasterol, b-sitosterol and D5-avenasterol. The distribution
of covalently bonded long chain fatty acids (acyl components) is very similar to that
of free fatty acids, suggesting that these free fatty acids are originated from the
hydrolysis breakdown of the bonded fatty acids.7-10

Phospholipids
Phospholipids in natural rubber are normally below 0.6% w/w. A recent
9-10
study showed that the major components were phosphatidyl choline and
lysophosphatidyl choline. Other minor components were phosphatidyl ethanolamine,
phosphatidyl inositol, lysophosphatidyl inositol, and metal phosphatidates or
phosphatidic acid. Unlike neutral lipids and glycolipids, the acyl components of
phospholipids normally contain very low level of furanoic acid, except for certain
rubber clones. The isolated fatty acids are mainly palmitic, stearic, oleic and linoleic
acids.7-10
Hydrolysis of phospholipids in the presence of ammonia produces phosphate
which could increase the stability of ammoniated latex during storage. Out of
phosphatidyl choline, phosphatidyl ethanolamine and phosphatidic acid, phosphatidyl
ethanolamine was found to be the most effective antioxidant.21

Proteins, amino acids and other nitrogenous compounds in NR

Proteins
About one quarter of natural rubber proteins are adsorbed on the surface of
rubber particles. Most of the adsorbed proteins are found in cream fraction of
centrifuged latex.22 The major proteins have the molecular weights of 14.5 and 29
kDa, which are similar to Rubber Elongation Factor and Hevamines, respectively.23
Half of the proteins are in the serum fraction.22 The molecular weight distributions of
these proteins are wide, ranging from 6 to more than 200 kDa. The remaining proteins
are found in the bottom fraction of centrifuged latex. The rubber particles in the skim
latex, on the other hand have a very low level of adsorbed proteins.2,3
The major protein in the serum of natural rubber latex is low sulphur -
globulin, having an isoelectrical pH of 4.5.24-26 In contrast to -globulin, hevein with
an isoeletrical pH of 4.7-4.9, is a cystine-rich protein having low surface activity.27-29
It is heat stable due to the presence of disulfide groups in cystine. Hevein has also
been reported to have the antifungal properties.28,29 Other proteins reported in the
literature include microfibrils, basic proteins such as hevamines A and B,24,30 some
high isoelectrical pH proteins31,32 and various types of enzymes.33 The rubber
biosynthesis enzyme was initially thought to be rubber particle bound. However,
recently, some enzymes from the bottom fraction have been reported to catalyze the in
vitro biosynthesis of rubber.34,35 In the case of high ammonia latex concentrate,36 the
molecular weights of five proteins have been found to be in the range of 14k-45kDa
and one greater than 100kDa. The major proteins that associated with the rubber
particles were found to be 14kDa while the minor 24kDa.
The carboxyl groups of proteins adsorbed on the surface of rubber particles
provide negative charges to the particles, contributing to the colloidal stability of the
latex.37,38 A recent Atomic Force Microscopy (AFM) study revealed that the majority
of the rubber particles surface was covered with proteins while the minority
phospholipids.39 Some of the proteins have also been reported to function as an
antioxidant40-44, e.g. glutathione which is tripeptide consisting of glutamic acid,
cysteine and glycine.45-46 Oxidizing agent can react with the –SH group of cysteine of
the glutathione. The rubber proteins have also been reported to promote storage
hardening of natural rubber, i.e spontaneous crosslinking reactions during storage,47,48
The storage hardening has been shown to contribute to the Plasticity Retention Index
(PRI) value of natural rubber, a measurement of aging resistance of natural rubber.49,50
Natural rubber proteins can also increase the moisture absorption,51-53 and high
modulus,54-56 creep and stress relaxation.51 Certain water soluble proteins in various
dipped products have been reported to cause Type I hypersensitivity (IgE mediated
allergy) in certain sensitive individuals.57-61 So far, 13 allergenic proteins have been
identified. Of these, Hev b 1, 3, 5, and 6.02 have been found to correlate with the
validated human IgE-based ELISA-inhibition test.62 Although most manufacturers
have taken steps to reduce the water soluble proteins in their products, as part of the
risks management, more users are now switching to synthetic rubber products,
particularly when the one is not sure of if a patient is allergy to the rubber proteins.

Amino acids
Fresh natural rubber latex contains about 0.1% of amino acids of which 80%
are in the serum fraction.63,64 The major amino acids in the serum of latex by weight
are aspartic acid, glutamic acid and alanine.45,65 Other amino acids include glycine,
phenylalanine, tryptophan, leucine, isoleucine, lysine, cystine. In addition, some trace
amount of valine, arginine, proline, serine and threonine have also been detected.63,64
Several amino acids such as histidine, glutamic acid, and glutamine, have
been found to show antioxidant activities.65 In addition, cystine, alanine, asparagine,
and phenylalanine have been reported to function as anti-oxidants in the study of
radiation vulcanized natural rubber latex (RVNRL).66 The reaction mechanism of
these amino acids at high temperature during vulcanization has also been reported.67
In the presence of amino acids, purified natural rubber was observed to undergo
storage hardening, indicating that the reactions involve amino acids.47
Other nitrogenous compounds
Apart from protein and amino acids, there are other nitrogenous compounds in
the latex. These compounds are mainly amines and the corresponding derivatives,
such as choline, methylamine, tetramethylenediamine, pentamethylenediamine,
ethanolamine, and trigonelline.30 These amine compounds, such as choline, a
quaternary saturated amine, have been reported to be an accelerator, and antioxidant
for raw rubber.68

Inositols and Carbohydrates

Quebrachitol (2-O- methyl L-inositol) constitutes about 1% of fresh natural rubber
latex.69 The purified compound has a peak melting temperature of 188-189oC.70 The
chiral structure of this compound allow one to simplify the syntheses of many drugs71-
73
The main glucid or saccharid in the latex is sucrose. In addition, a small quantity of
glucose, galactose, fructose, raffinose and two pentoses has also been reported.74-76
Microbial attack on carbohydrates can lead to the generation of volatile fatty acids
(VFA).77 A low VFA number has been found to give a better physical property to the
film either un-vulcanized or vulcanized, vice versa.78

Ash
The major components of natural rubber ash are phosphorous, potassium, sodium and
magnesium, while the minor components copper, manganese, iron, and calcium.31, 79-83
A high magnesium level is known to cause low mechanical stability and high volatile
fatty acids in latex.84 On the other hand, phosphates are known to give good stability
property to the latex. Therefore, the ratio of phosphate to magnesium ratio is
important in determining the stability of the latex.85-88 The phosphate can form an
insoluble precipitate with the magnesium ions.89-91 During storage in ammonia, the
hydrolysis of phospholipids further releases more phosphates. Therefore, the latex
mechanical stability increases with storage time.
Copper, manganese, and iron can catalyse the oxidative degradation of natural
rubber, with copper being most active among the three.92 However, only free transition
metals can act as a catalyst in the rubber oxidation process.93 When in latex, these
metals might be in complex form and do not catalyse the oxidative degradation
reaction. These metals may be released when their complexes undergo chemical or
biological reactions.6
The spontaneous crosslinking reaction of natural rubber has been postulated to
involve the multivalent metal ions such as calcium and magnesium and carboxylic
acids of bonded proteins or amino acids.48 Although the addition of these metal ions
did not lead to the hardening of the rubber,94 a microscopic analysis on the gel fraction
of natural rubber by transmission electron microscopy associated to electron energy-loss
spectroscopy revealed the presence of ionic crosslinking.95 This seems to contradict the
conclusion from an early study using deproteinization, saponification, and centrifugation
to obtain the purified rubber, which is rather non-specific.96,97

Volatile Matters
Moisture is the main component in the volatile matters. There are small quantities of
other volatile acids such as formic, acetic, and propionic in the latex.98 The moisture
retention of natural rubber is enhanced by the presence of hydrophilic impurities
mainly inorganic salts and to some extent proteins.53 A high moisture content can
promote mold growth and cause undesirable odor to the dry rubber.98
Future trend
Quebrachitol is probably the only non-rubber that has attracted the attention of
scientists for commercialization. The non-rubbers are currently either being discarded
or recovered for low value applications such as fertilizers99 and Bifidobacterium
bifidum growth stimulator.100 The extraction of potentially high commercial value
non-rubbers such as glutathione and tocotrienols have not been fully explored,
probably because of their low concentrations in the latex, making it uneconomical to
extract. With the development of genetic engineering, such high value non-rubbers
content may be boosted.101-103

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