National Dairy Research Institute, NDRI, Karnal

Course Title: Advances in chemistry of milk processing (DC-721) Date: 07 Feb’11

Name of the student: Minaxi
Class: Ph.D, I year, II sem (DC)

ASPARTAME AND ACESULFAME K- THE ARTIFICIAL SWEETNERS

Introduction
In the last three decades, the growing concern about health and life quality has
encouraged people to exercise, eat healthy food and decrease the consumption of food rich in
sugar, salt and fat. In addition, there has been an increase in the demand by consumers for food
with functional properties, resulting in the concept of production of quality food. More recently,
there is increased consumer interest in reducing sugar intake. Food products made with
sweeteners rather than sugar have become more popular. The discovery of a great number of
sweeteners during the last decade has triggered the development of new sugar-free products,
particularly for diabetics, people on special diets and/or for the obese.
Sweeteners are alternative substances to sugars, which give food a sweet taste and are
used to partially or totally replace sucrose. During the past few decades, low-calorie artificial
sweeteners, such as aspartame, saccharin, acesulfame-K and sucralose, have become sugar
alternatives to replace sucrose, and have been widely used in dairy products especially in Europe
and USA (http://www.informaworld.com/smpp/title~content=t713401167).
Today aspartame is an integral part of the modern diet and is sold commercially under
names like NutraSweet and Canderel, aspartame can be found in more than 5,000 foods,
including fizzy drinks, chewing gum, table-top sweeteners, diet and diabetic foods, breakfast
cereals, jams, sweets, vitamins, prescription and over-the-counter drugs.
Because it contains no calories, aspartame is considered a boon to health-conscious
individuals everywhere. But independent scientists say aspartame can produce a range of
disturbing adverse effects in humans, including headaches, memory loss, mood swings, seizures,
multiple sclerosis and Parkinson's-like symptoms, tumours and even death
(www.theecologist.org.).
ASPARTAME
Description
Aspartame, a synthetic nonnutritive sweetener, is a methyl ester of a dipeptide composed
of aspartic acid and phenylalanine. There are two forms of aspartame, an alpha and a beta form.
Only α form is sweet and aspartame generally refers to the α form. It is a white crystalline
powder having no odor, but is intensely sweet, approximately 200 times sweeter than sucrose
(the accepted standard for sweetness). Although aspartame yields the same caloric intake as
sugar on a weight to weight basis (i.e., 4 kcal/g), it can be added at almost 200 times lower levels
and achieve the same sweetness, thereby providing a far lower net caloric intake. This attribute
has resulted in the use of aspartame as a low calorie or nonnutritive sweetener in foods and
beverages worldwide (http://www.informaworld.com/smpp/title~content=t713401167).

Chemically, this sweet-tasting compound is N-L-α-aspartyl-Lphenylalanine-1-methyl

ester, and may be synthesized from a mixture of compounds by chemical reaction or by the
action of enzymes with high specificity. It consists of 39.5% aspartic acid, 50% phenylalanine
and 10.5% methyl ester (Pinheiro et al., 2005) having a molecular weight of 294.30 and formula
C14H18N2O5 (http://www.dorway.com/help.txt). Although the individual amino acids have no
sweetness, but when combined, make the product 200 times sweeter that sucrose.

History of Use:
Aspartame was discovered accidentally in 1965 by James Schlatter, a scientist in the G.
D. Searle research laboratories, (that later became a Monsanto company), while working to
synthesize a tetrapeptide as an inhibitor of the gastrointestinal secretory hormone, gastrin. While
preparing the tetrapeptide, he was working to crystallize an intermediate (a dipeptide),
aspartylphenylalaninemethyl ester, and some of the solution accidentally was spilled onto his
hands. Against all good safety practices, Schlatter licked his fingers to pick up a piece of paper
and discovered the intense sweet taste of the intermediate
((http://www.informaworld.com/smpp/title~content=t713401167).
Cloninger and Baldwin (1970) published a report in Science in 1970, comparing the
sweetness and other flavor attributes of aspartame in comparison with sugar and other
sweeteners (saccharin and cyclamate) and proposed its use as an artificial sweetener. In 1980 it
was approved as a food additive alone or in combination with other sweeteners, it was introduced
in the U.S. in 1981, followed by in 1991 approval as a tabletop sweetener. It is best known to the
public by the trade names NutraSweet® and Equal® (http://www.dietriffic.com/).

Structure:
The aspartame molecule consists of three substances linked together: the amino acids,
phenylalanine and aspartic acid, and methanol (methyl alcohol). The two amino acids present in
aspartame are found naturally in foods as protein components. Methanol also occurs naturally in
some foods and is produced by the digestion of other food constituents but aspartame itself does
not occur naturally.

Phenylalanine Aspartic Acid Aspartame

Figure: Phenylalanine and Aspartic Acid to yield Aspartame

Manufacturing Process
Aspartame is manufactured by the coupling of amino acids L-phenylalanine methyl ester and L-
aspartic acid to produce the dipeptide methylester. If the coupling is done chemically, both the
sweet α form and the nonsweet β form of aspartame are produced, requiring separation to obtain
only the α form. The enzymatic process yields only α-aspartame. Esterified L-phenylalanine
(Phe) is reacted with Nprotected L-aspartic anhydride (N-protected-Asp O) to form Nprotected
alpha-aspartame. After the coupling reaction, a series of steps of protection removal,
crystallization, decolorization, filtration, sterilization, fine crystallization, and drying, the final
product aspartame (L- aspartylphenylalanine methyl ester) is obtained.

Fig. Flow chart for the production of aspartame.

The individual amino acids are produced by an initial fermentation process using specific
strains of bacteria. Aspartic acid is made from the bacteria Brevibacterium flavum, while the
phenylalanine is generated from Corynebacterium glutamicum. These two materials are
combined through amidation to establish the peptide bond.
Metabolism of aspartame:
One of aspartames major health risk comes from the reverse of this process in the body.
Once ingested in the body, it is metabolized in the gastrointestinal tract by esterases and
peptidases to methanol and the two amino acids.
Aspartame may be completely hydrolyzed to these three components in the
gastrointestinal lumen and absorbed into the general circulation, or may be hydrolyzed to
methanol and aspartyl-phenylalanine dipeptide. In this case, the dipeptide is absorbed into the
gastrointestinal mucosa cells and then cleaved into amino acids. Aspartame can also be absorbed
into the mucosal cells prior to hydrolysis, and be cleaved within the cell, to its three components,
which then enter circulation. The available evidence indicates that aspartame does not enter the
circulation prior to hydrolysis. Metabolism of the three cleaved components of aspartame has
been shown to be identical to the metabolism of the components given individually.
The extra addition of phenylalanine only bothers those with a specific disorder (a rare
genetic disorder phenylketonuria), which means they cannot handle certain concentrations of
phenylalanine (Phe). Individuals with PKU lack the oxidizing system, specifically Phe-
hydroxylase, that converts excess Phe into tyrosine (Tyr) through “transamination with α-
ketoglutarate to p-hydroxyphenylpyruvate” and then ultimately into homogentisic acid.
Excess Phenylalanine accumulated in the blood can lead to “metabolic acidosis”,
distortions of plasma concentrations of other amino acids which can affect brain function due to
lack of neurotransmitters, in particular serotonin. Even with the health risks associated with
aspartame, it is used in a multitude of food applications, including tabletop portions, with the
exception of baked goods as degrades at temperatures above 90°F; although there have been
recent advances to encapsulate it in order to protect it under heated conditions
(http://www.dietriddic.com).

Figure: Metabolic processing of phenylalanine

A fairly complete list of aspartame breakdown components from the aspartic acid, phenylalanine,
and methanol is as follows:
 Formaldehyde CH2O  Norepinephrine
 Formic Acid CH2O2  Epinephrine
 Beta Aspartame  Phenylethylamine
 Aspartylphenylalanine  Phenylpyruvate
 Aspartylphenylalanine amide  Phenylactic acid
 Tyrosine C9H11NO3  Phenylacetic acid
 L-Dopa  Diketopiperazine (DKP)
 Dopamine
(http://www.dorway.com/help.txt)
Estimated daily intake
The evaluation of safety of adding a food component to the food supply requires
consideration of the amount of the component that will be added to food, the amount of its
metabolites that will be in the foods, and the amount of these components that are already
present or naturally occurring in the food supply.ADI is 50 mg/kg body weight.
Characteristics:
Aspartame is a white powder, and does not leave any bitter, chemical or metallic
aftertaste. Its sweetness is 133 and 200 times greater than that of sucrose at 4 and 10%,
respectively (Pinheiro et al., 2005). Aspartame provides four calories per gram — as many
calories as protein or sugar. Aspartame can be stored for prolonged periods in a dry state, both
pure and blended with other ingredients.

Technical data Aspartame

Molecular formula C14H18N2O5

Molecular weight 294

Appearance Odourless, crystalline white powder,
with a strong sweet taste
Function Sweetener
Sweeteness intensities 133-200 times sweeter than sucrose
Solubility Slightly soluble in water and ethanol

Properties:

1. Stability:
The stability of aspartame is affected by time, humidity, pH (neutral or alkaline) and by
very high temperatures for long durations (e.g. cooking, baking or sterilization), but not by its
concentration.

Storage condition:
Aspartame is very stable under dry conditions, but degrades during prolonged heat
treatment in aqueous solutions. A study identified five degradation products of a 1% aqueous
◦
solution of aspartame following storage at 37 C for 2 months at pH 4.6. The compounds were 3-
carboxymethyl-6-benzyl-2,5- diketopiperazine (DKP), L-aspartyl-phenylalanine, L-aspartic acid,
L-phenylalanine, and L-phenylalanine methyl ester. None of the breakdown products are sweet
(Maisons-Alfort , 2002).

pH and Temperature:
The rate of degradation in aqueous solutions depends on the pH and the temperature.
Aspartame is most stable between pH 4-5, with a half-life of over 250 days at 25◦C.
At room temperature its stability is good at pH values of between 3.4 and 5 and it is
maximum at pH 4.3.
At pH below 3.4 the dipeptide is hydrolysed and at a pH greater than 5, cyclisation occurs
with the formation of diketopiperazine (DKP) or hydrolyze to α-aspartylphenylalanine and
methanol (DKP and α-aspartylphenylalanine can interconvert, but do not revert to aspartame). In
both cases, this transformation results in the loss of sweetness.
 At acidic pH (less than 4.5), aspartame can undergo rearrangement to β-aspartame.
(http://www.informaworld.com/smpp/title~content=t713401167).

Figure: Effect of pH on aspartame

Aspartame is very stable in the dry state. At 105°C a loss of approximately 5% (due to
formation of diketopiperazine) is observed after 100 hours of treatment. At 120°C, a 50% loss is
obtained after 80 hours of treatment. In solution, when stored at temperatures ranging from 30 to
80°C, aspartame is progressively degraded into diketopiperazine (Maisons-Alfort , 2002). It is
therefore not usable in foods heated at higher temperature (cooking, sterilization, etc.)
(http://www.informaworld.com/smpp/title~content=t713401167).

2. Solubility:
The solubility of aspartame in water is dependent on pH and temperature. The solubility
in water is low (1 g/ 100 g at 25°C), but increases as the pH is decreased to 2.2 or by increasing
the temperature to 40°C at pH 4. Aspartame is insoluble in oil or fat (Pinheiro, et al., 2005).
The maximum solubility is reached at pH 2.2 (20 mg/ml at 25°C) and the minimum
solubility at pH 5.2 is 13.5 mg/ml at 25°C (Maisons-Alfort, 2002).

3. Sweetening power:
The sweetening power of aspartame is generally greater at low concentrations and in
products stored at room temperature. It enhances the aroma and prolongs the perceived taste of
fruit, mainly the acidic fruits (Pinheiro, et al., 2005).

4. Safety:
Methanol is dangerous as inside cells, it is converted to formaldehyde, an undetectable
toxin and recognized cancer causing agent, rated at the highest order (Group I) by the
International Agency for Research on Cancer (IARC). Even when formaldehyde is injected
directly into a living human, it turns into formalhydrate, an aggressive molecule that instantly
attaches to any protein molecule with which it makes contact. The formaldehyde molecule
completely disappears under the cover of the much larger protein, which then loses function. No
diagnostic procedures can detect a protein molecule so changed. Damaged protein molecules are
not tolerated by the immune system. Specific detection sites for ‘formaldehyde-modified protein’
are found on white blood cells called macrophages. Macrophages seek out and destroy these
proteins at a rate 100 times faster than they do with proteins not treated with formaldehyde
(www.fitnesslife.co.nz).

APPLICATIONS:
Aspartame is used in a wide variety of foods and beverages and as a table-top sweetener.
Aspartame is unstable if subjected to prolonged heating, so it is not suitable for use in cooking or
baking. It also breaks down in liquids during prolonged storage. However, control of the
distribution system has allowed aspartame to be used successfully in liquid products including
carbonated beverages (http://www.informaworld.com/smpp/title~content=t713401167).
The aspartame can be used at a rate 0.028 g/100 g of yogurt (Pibheiro, et al., 2005).

ACESULFAME-K

High intensity sweeteners like acesulfame K (Sunett™) are used as a partial substitute for
sucrose in most sugar free or reduced calorie hard candy formulations and contribute to the
overall desired sweetness. Due to their intense sweetness only small quantities are used which do
not supply bulk and do not contribute to the textural characteristics of the hard candy. Therefore,
intense sweeteners must only be regarded as sweet tasting compounds or sweet flavoring
compounds in hard candy production (Nakhost, 1995).
Acesulfame K (synonyms: Acesulfam K; Potassium salt of 6-methyl-1,2,3-oxathiazine-
4(3H)- one-2,3-dioxide; Potassium salt of 3,4-dihydro-6-methyl-1,2,3-oxathiazine-4-one-2,2-
dioxide; Acetosulfam) is an artificial sweetener, approximately 200 times sweeter than sugar. It
is a white, odourless, crystalline powder.
It is approximately 130-200 times sweeter than sugar. Because
acesulfame-K is heat-stable, it can be used in cooking and baking.
Acesulfame-K is not broken down in the human body, and therefore it
provides no calories. It does not promote tooth decay.

History:
Acesulfame-K was discovered by accident by German scientists Clauss and Jensen in
1967, while they were conducting research on new cyclic group, dihydro-oxathiaxinone dioxide.
(http://www.dietriffic.com/).
Acesulfame-K first approved for use in foods in the United Kingdom in 1983. In 1988,
the Food and Drug Administration (FDA) approved the use of acesulfame-K in a variety of dry
food products. It is now approved in confections, baked goods and soft drinks as well and is now
approved for various uses in more than 90 countries. In the U.S., in 1992, it began to appear on
the market, under the trade names Sunette ™ and Sweet One™. In Europe, acesulfame- K is
already in use in a variety of products

Properties:
Physical and chemical property:
Appearance: Free flowing colourless crystals or white, crystalline powder
Good taste: Similar to sucrose in terms of taste (at high doses, it has a slight aftertaste)
High sweetness: About 200 times sweeter than sucrose
Solubility: Dissolves quickly and easily in water under room temperature
Non-calories: No metabolism and no calorific value in the body

Molecular weight: 201.24

Chemical Formula: C4H4KNO4 S
Pure/Mixture: Pure
Solubility in H2O: Appreciably soluble
Solubility others: Slightly soluble in ethanol

Thermostability: Highly thermostable (therefore suitable for baking and cooking)

Synergistic effect: Acesulfame-K has a distinguished synergistic effect when used in
combination with other sweeteners, which can make a 20-40% reduction
in the quantity of product required
(best synergy results with Aspartame and Cyclamate)
Shelf life: extended shelf life
Production cost: low production cost (Miltnberg and Samton, Inc.).

Effect of pH and temperatute: Under extreme conditions of pH and temperature, detectable

decomposition may occur leading to the formation of acetone, CO 2, and ammonium hydrogen
sulfate, or amido-sulfate, as final decomposition products; under acid (pH 2.5) conditions,
minute quantities of acetoacetamide and acetoacetamide N-sulfonic acid are formed as unstable
intermediate decomposition products, while under alkaline (pH 3-10.5) conditions, acetoacetic
acid and acetoacetamide N-sulfonic acid can be detected.

Physiological characteristics:
It is not converted in the body and is excreted unchanged by the kidneys, hence is calorie-
free, tooth friendly and suitable for diabetics.
ADI-value:
15 mg per kg of body weight.

Safety:
Since, acesulfame-K is not metabolized and does not accumulate in the body, that it has
no drug-like effects, and that it does not cause cancer or birth defects, hence doesn't affect the
normal function of human body.
After studies, FDA concluded that acesulfame-K is safe for its intended uses when level
of consumption is well within the acceptable range.
In February 1992, FDA concluded that, “Acesulfame potassium has been thoroughly
tested for safety... the agency has concluded that the studies conducted to establish the safety of
this compound are adequate to demonstrate, to a reasonable certainty, the safety of acesulfame
potassium for its intended uses”.
After this, Acesulfame-K has also been scrutinized and approved by international
regulatory bodies including the Scientific Committee for Foods of the European Community and
the Joint Expert Committee on Food Additives (JECFA) of the World Health Organization
(WHO) and the Food and Agriculture Organization (FAO) of the United Nations (Kathleen et al.,
1993).
Joint Expert Committee on Food Additives. reviewed extensive toxicological studies on
the breakdown products, acetoacetamide and acetoacetamide-N- sulfonic acid, which indicated
that these compounds have a low toxicity and are not mutagenic.

Thus, scientists and regulatory authorities have concluded that even under extreme
acidity, temperature and storage conditions, acesulfame potassium has been found to be safe for
use in beverages.

Manufacturing Process:
Acesulfame potassium is made from a process that involves the transformation of an organic
intermediate, acetoacetic acid, and its combination with the naturally occurring mineral,
potassium, to form a highly stable, crystalline sweetener (http://ificinfo.health.org).

Application:
A major positive trait is its stability under heat applications. This translates into longer
shelf life, which makes it highly suitable in packaged goods, as well as available for use in
baking.
It is used in chewing gum, dry beverages mixes, instant coffees, teas, gelatin, puddings,
non-dairy creamers, water- and milk-based drinks puddings, desserts ice cream and frozen
desserts toothpaste, mouthwash pharmaceuticals etc.
Application has been made for its use in carbonated beverages, baked goods, and hard
and soft confections. It is used in 1000 food products in worldwide.

USE IN BLENDS

Acesulfame potassium is often used in sweetener blends to produce a more sugar-like taste in a
food or beverage. The ingredient also helps the blend retain its sweetness during baking or heat
processing, which is important for preparing foods, such as cookies and candies. Acesulfame
potassium helps blends sustain their sweetness over time, thereby increasing the sweetness shelf
life of products. In addition, foods containing blends of acesulfame potassium contain up to 40
percent less total sweetener.