Method For Preparation of Vitamin C and Method For Determination of Vitamin C in Tablets
Method For Preparation of Vitamin C and Method For Determination of Vitamin C in Tablets
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Head - QC, Qa/R&D-Ipl Research centre, Lucknow, India
Abstract
Procedure for calcium ascorbate (VITAMIN-C) includes such technological steps as reaction of ascorbic acid
on calcium carbonate in water in recent years; the determination of vitamin C has become an important subject in the field
of biochemistry and commercial foods. This is because vitamin C plays an important role in maintaining human health.
Due to the importance of vitamin C in human beings, the quantitative analysis of vitamin C has gained a significant
increase in several areas of analytical chemistry such as pharmaceutical and food applications there are numerous
methods for the determination of vitamin C in a variety of natural samples, biological fluids and pharmaceutical
formulations. The preparation method and methods for the determination of vitamin C are spectrophotometric
methods and non-spectrophotometric methods. For non-spectrophotometric methods are such as high-performance
liquid chromatography (HPLC), titration, enzymatic method and fluorometry. Direct spectrophotometry also has been
applied to determine the vitamin C content in soft drinks, fruit juices, and cordials after correction for background
absorption in the UV region.
Received June 05, 2018; Accepted June 21, 2018; Published July 04, 2018
Empirical formula: C6H8O6
Citation: Krishnasarma Pathy (2018) Process for Preparation of Vitamin C
Molecular weight: 176.1 and Method for Determination of Vitamin C in Tablets. SF J Chem Res 2:1.
Melting point: about 190°C (with decomposition) Copyright: © 2018 Krishnasarma Pathy. This is an open-access article
distributed under the terms of the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any
Appearance: white to slightly yellowish crystalline medium, provided the original author and source are credited.
powder, practically odorless, with a strong acidic taste.
The water-soluble vitamin C is probably the most acid. Vitamin C is probably one of the most highly well
well-known vitamin. Even before its discovery in 1932, known. Furthermore, people have become more aware to
physicians recognized that there must be a compound the importance of vitamin C. Hence, this causes the global
in citrus fruits preventing scurvy, a disease that killed market flooded with vitamin C fortified foods. The term
as many as two million sailors between 1500 and 1800. of vitamin C is used as generic term for all compounds
Later researchers, discovered that man, other primates and exhibiting qualitatively the biological activity of ascorbic
the guinea pig depend on external sources to cover their acid. The molecular structure of vitamin C is C6H8O6 and
vitamin C requirements. Most other animals are able to the molecular weight is 176.1. Vitamin in foods: Analysis
synthesize vitamin C from glucose and galactose in their Bioavailability and Stability. United States of America:
bodies. CRC Press Taylor and Francis Group. Vitamin C is highly
Nowadays, health has become the most important polar and readily soluble in aqueous solution and insoluble
property of human’s life. Commonly, diets with high in less nonpolar solvents. It is an acidic compound due to
contents of fruits are protective against several human the facile ionization of hydroxyl group on carbon 3 (pK1
diseases such as cardiovascular diseases and even cancer. = 4.17) while the hydroxyl group on carbon 2 is much
Therefore, people are putting more and more attention on more resistant to ionization (pK2 = 11.79). The structure
antioxidant substances such as vitamin C which is also of L-ascorbic acid is shown in Figure 1.
known as ascorbic acid or more specifically L-ascorbic
Figure 1: Schematic Diagram of the rFIA-CL Manifold used in this Study for the Determination of Ascorbic Acid
Ball also stated that ascorbic acid is easily and vitamin E. It is also found to be a strong antioxidant as
reversibly oxidized to dehydroascorbic acid, forming it helps to neutralize harmful free radicals. Iodometric
the ascorbyl radical anion which is also known as Determination of Ascorbic Acid(Vitamin C) in Citrus
semidehydoascorbate as an intermediate as shown in Fruits. Research Journal of Agriculture and Biological
Figure 2. Dehydoascorbic acid possesses full vitamin C Sciences.
activity because it is readily reduced to ascorbic acid in the Vitamin C is an almost odorless white or pale
animal body. However, dehydoascorbic acid is not an acid yellow crystalline powder with a pleasant sharp taste and
in the chemical sense, as it does not have the dissociable melting point of about 190 and deg;C. It is not a carboxylic
protons that ascorbic acid has at carbon 2 and carbon 3 acid but a lactone and ease of oxidation to the presence of
positions. an enediol grouping [5]. Vitamin C is highly susceptible
One of the most important properties of vitamin C to oxidation, especially when catalyzed by metal ions
is that it is an antioxidant. Nevertheless, it has a wide range such as copper(II) ion and iron(III) ion. The functions
of antioxidant properties outside the body and can quench and activities of vitamin C are based on its properties as a
most biologically active radicals. It scavenges superoxide, reversible biological reductant. Image 1
nitroxide, hydroxide, hydrogen peroxide and will reduce
1. Preparation of Iodine Solution were weighed and smashed to form powder and average
For preparation of 0.1 M iodine solution, 10g of value was calculated.
KI was taken in a 250 ml Volumetric flask and 35 ml of
distilled water was added followed by heating the solution; Equipment and Apparatus 2
the mixture was cooled to room temperature and 3.15 g of 334 mg of the powder tablet was taken in a 100.00
solid Iodine powder was dissolved. Similarly, to prepare ml beaker and dissolved in 100 ml distilled water. 5.
0.005M of iodine solution 2g of KI was taken in a 500 ml Standardization of the iodine solution with the vitamin
beaker and dissolving in 100 ml of distilled water and 1.3 C standard solution and sample solution. The measured
g of iodine powder was stirred with small quantity of water volume of 20ml of both standard and sample was taken
and qs (quantum satis) to 1 liter. from each solution and equilibrated with 150 ml distilled
water separately into distinct two Erlenmeyer flask 250.00
2. Preparation of Starch Solution ml and titrant containing iodine solution was run against
Addition of 0.25 g of starch powder in 50ml warm analyte containing either sample or standard; 5-6 drops
distilled water, As the starch is insoluble in cold water and of prepared starch solution were added to the analyte and
needs to be boiled to stay in solution. titration was started. The burette level for each analyte for
distinctive experiment was noted as mentioned below:
3. Preparation of Vitamin C Standard Solution For standard solution the volume of iodine solution
25mg Ascorbic acid was taken in a 100.00 ml required for complete reaction = 45 ml Equally, for
beaker and dissolved in 100 ml distilled water. Sample solution the volume of iodine solution required =
49 ml The endpoint was noted when analyte appears blue
4. Preparation of Vitamin C Sample Solution in color.
From the strip of Vitamin C random two tablets
Equipment and Apparatus 2:
C where its systems allow faster sampling rates and pharmaceuticals by flow injection analysis using brown
consumed fewer reagents compared with segmented- mono 1-10-Phenanthroline-Iron(III) complex as oxidant.
flow analysis. Memon, Dahot and Ansari had proposed Medical Journal of Islamic Academy of Sciences.
a method by using mono 1, 10-phenanthroline-iron(III) This method can be improved within certain limits
complex as oxidant. This experiment was based on its by increasing the volume of the injected sample in flow
reducing reaction on mono(1-10-Phenanthroline)-iron(III) injection analysis. The sensitivity is increased two fold with
to tris(1,10-Phenanthroline)-iron(II) (ferroin) and the the increase of sample volume. As conclusion, since the
absorbance of ferroin was monitored at 510nm through time required for sample preparation is short and reagent
spectrophotometer equipped with a flow through cell. consumption is low, hence the method is highly economical
In this analysis single channel manifold is used as and is suitable to use on routine basis for determination of
shown in Figure 2. The reagent stream is pumped at the ascorbic acid in pharmaceutical preparations.
flow rate 1.1mL/min via a peristaltic pump equipped with
PVC pump tubing. The vitamin C sample is introduced into Ultraviolet (UV) Spectrophotometry
the reagent stream via a rotary teflon valve. A calibration Direct ultraviolet spectrophotometry is a fast,
curve for vitamin C in the range 0-50ppm was plotted from simple and reliable method for the determination of
the results obtained by Memon, Memon, Dahot and Ansari vitamin C. This method can be done through alkaline
which are shown in Figure 2. treatment and the maximum absorption of vitamin C falls
Figure 2: Reagent Concentration (%) at 243nm at pH2. The absorption of UV light by the sample
matrix was the major problem in this method. Therefore,
alkaline treatment method was found to be used as
background correction in blank. This is because more than
95% of vitamin C will be destroyed in 10 minutes after
alkaline treatment which is in the range of pH 12 to 13.
UV spectrophotometry method was found to be applicable
for most fruits, fruit juices and soft drinks except those
that are unstable to alkaline treatment, and were deeply
colored, or contained high concentration of caffeine,
saccharin, caramel and tannic acid. Study of Direct
Figure 3:
Ultraviolet Spectrophotometry on the Determination of
Ascorbic Acid In Fruit Juices and Soft Drinks. Journal of
China Agriculture University.
To determine the total content of vitamin C in
food samples, a well-established method was investigated
by Khan, Rahman, Islam and Begum, 2006 by using the
2,4-dinitrophenyl hydrazine methods (DNPH). This is
a simplified method for the simultaneous determination
of total vitamin C employed coupling reaction of
2,4-dinitrophenyl hydrazine dye with vitamin C and
followed by spectrophotometric determination. The
spectrophotometric method involves the oxidation of
ascorbic acid to dehydroascorbic acid [4, 10] by the action
They also studied about the effect of reaction of bromine solution in the presence of acetic acid. Reaction
coil and reagent concentration. From the graph (Figure between dehydoascorbic acid and 2,4-dinitrophenyl
3), the maximum intensity was observed at 50cm hydrazine at 37 and deg;C temperature for three hours
reaction coil. While the results of the effect of reagent will form an osazone. The solution is treated with 85%
concentration obtained is shown in Figure 6 indicating that H2SO4 to produce a red color complex. The absorbance of
the maximum signal could be obtained at 35% reagent. all standards was measured at 521 nm (a wavelength range
Spectrophotometric determination of ascorbic acid in from 350 nm to 650 nm showed a maximum absorbance
((ƛmax) at 521nm) by using a UV-spectrophotometer. The form osazone when react with DNPH [11-14]. Hence,
results obtained were taken to contruct a calibration curve. there is a chance of error in this method which may give
Figure 3.1 false results.
Figure 3.1: Another interference was due to the extracted
glucose which contains similar structure like vitamin C.
Therefore, some of the glucose may be extracted in the
meta-phosphoric acid during the extraction of ascorbic
acid from sample. Glucose may also cause the formation
of colored complex with DNPH and gives the false result
in the determination of vitamin C. This was proven in
Figure 4.1 where there is no absorption peak around the
interested peak at 521nm.
Figure 4.1: Calibration curve of standard vitamin C at 521 nm
Figure 4:
to prevent the formation of the quinoxaline derivative. bands at 682nm for MB and 452nm for LMB. In Figure
It is used to reveal any fluorescence due to interfering 7, the emission peak of MB at 682nm increased due to
substances. the increase of its concentration. A linear relationship
Figure 5: between MB concentration and intensity was obtained
over the concentration range of mol L-1 MB (y= 49.082x
+ 94.46,r2=0.9969). The excitation peak of MB at 664 nm
also linearly increased depending on the increase of its
concentration.
Figure 7:
6
mol.l-1
vitamin C of orange juice is readily oxidized and lost prepared which one contained vitamin E stock solution
during staying of the juice. On the other hand, there are and vitamin C stock solution while another contained only
several factors that will also affect the stability of vitamin vitamin C stock solution. The samples were analysed once
C in orange juice. The factors are such as the effect of a week for five weeks.
vitamin E, pH, and parameters which include air, heat, The results of the stability of vitamin C show that
water as well as prolonged storage and overcooking. the presence of vitamin E influenced the decay of vitamin
According to Ball, a meta-oxygen-ascorbate C. It shows that there were differences between samples
complex is formed in the presence of molecular oxygen with or without vitamin E, it can be clearly seen that the
and trace amounts of transition metal which particularly concentration of vitamin C without vitamin E fell down
are copper (II) and iron (III). This complex contains a to 1.2mg/L on the second day. However, in the presence
resonance form of a diradical that rapidly decompose to of both vitamins, the decay was also observed, but it was
give the ascorbate radical anion, the original metal ion, and lesser. The concentration of vitamin C in the orange juice
hydrogen peroxide. This radical anion will in turn reacts with vitamin E was 13mg/L in the fifth week.
with the oxygen to give dehydroascorbic acid (DHAA) As a result, it seems that vitamin E stabilized
[15]. For anaerobic pathway of vitamin C which occurs vitamin C in orange juice at a determined concentration.
in the absence of free oxygen, the degradation is caused This is because vitamin E delay the oxidation of vitamin
by the formation of diketogulconic acid. As the rate of C thus, enhances the stability of vitamin C in orange juice.
degradation is maximum at pH 3 to pH 4, therefore this The combination of vitamin C with vitamin E makes
pathway is mostly responsible for anaerobic loss of vitamin the orange juice more stable and slower the degradation
C in canned grapefruit and orange juices. of orange juice. This concluded that orange juice with
vitamin E addition is a good way to preserve the vitamin
Effect of Vitamin E on The Stability of Vitamin C C content during storage. The stability of vitamin C in
In Orange Juice different beverages. British Food Journal.
Vitamin E is a fat soluble antioxidant that has
four tocopherols and four tocotrienols. In nature, these Effect of Temperature on The Stability of Vitamin
four tocopherols and four corresponding tocotrienols are C In Orange Juice
designated as alpha-(?), beta-(?), gamma-(?) and delta-(?) Vitamin C of fruit juice is readily oxidized
according to the number and position of methyl substituent and lost depends on the conditions of storage. There
in chromonal ring. are studies about the determination of the amounts of
The vitamin E functions as a biological antioxidant vitamin C content in fruit juices under different storage
by protecting the vital phospholipids in cellular and conditions. Kabasakalis, Sipadou and Moshatou had done
subcellular membranes from peroxidative degeneration. an experiment to determine the rate loss of vitamin C
Vitamin E mostly accumulates in body which are liver with respect to time and temperature of storage. A long-
and pancreas. But unlike vitamins A and D, vitamin E is life and short-life commercial orange juice 100% without
essentially nontoxic. preservatives and fresh orange juice were used for analysis.
Nagymate and Fodor have designed a method to In this experiment, the days before the expiration date
study the effect of vitamin E on the stability of vitamin C. were recorded in Table 3. to observe the loss of vitamin
In this experiment, vitamin E stock solution was prepared C in short-life and long-life orange juice 100% as the
by dissolving ?-tocopherol in absolute ethanol. The orange expiration date was approached.
juice which contained vitamin E and vitamin C was
used as sample. The storage temperature of the vials was Table 3: Program of the Temperature conditions for sample Storage
4anddeg;C and they were covered with aluminium foil STORAGE TEMPERATURE(0C)
to prevent the effect of sunlight. Besides, two different TIME (h)
Condition n0 1 Condition n0 2
temperatures were used to examine the effect of vitamin E
0 4 4
at that temperature which half of the samples were stored at
4 8 8
20anddeg;C. On the other hand, the additive effect of these
8 4 12
vitamins was also examined but only cool samples (4 and
deg;C) were used for this experiment. Two samples were 12/72 4 8
Table 3. shows the loss of vitamin C from fresh As reported, decreases of vitamin C upon storage did not
and long-life commercial orange juice 100% during a 31 correspond to increases of dehydroascorbic acid levels. In
days period, with measurements made every 1 to 3 days. fact, there was an increase of dehydoascorbic acid levels
The samples were refrigerated into containers which in aseptically packaged orange juices. This means that the
after the initial measurement remained either open or overall nutritional quality of orange juices is affected upon
with closed cap until the next measurement. Based on storage.
the results shown in table 4, the magnitude of vitamin C The loss of the vitamin C in a commercial long-
did not differ significantly between open and closed cap life orange juice 100% stored in refrigerator and non-
for both juices. The commercial orange juice lost higher refrigerated for a period of 10 days in open containers
amounts of vitamin C compared with fresh orange juice. were shown in Figure 11.
Table 4: Reaction Rate Constant (k) for Ascorbic Acid Degradation in the Presence of Hydrogen Peroxide in Various Fruit Juices during Storage
Zero-Order First-Order
Sample H2O2Concentration(ppm) Temperature
K(mgl1h1) R2 K (h-1) R2
0.5 20 1.24 0.972 0.0029 0.922
0.5 30 2.1 0.953 0.0058 0.854
Orange Juice
0.5 40 3.5 0.989 0.0131 0.885
5 40 6.62 0.957 0.0184 0.964
Figure 11:AA Concentration (mg/100g) in Natural Orange Juice Stored Under Isothermal Conditions (a) and Non- Isothermal (b) conditions
(Condition n0 1- 4h/80C and 60h/40C; Condition n02-4h/40C, 4h/120C and 64h/80C)
Table 5: Assessment of Unpasteurized Refrigerated Orange Juice Stored Under Isothermal and Non-Isothermal Conditions for 72 hours
According to Figure 2, Table 5 non-refrigerated samples increase the degradation of vitamin C. However, raising
show higher percentage loss of vitamin C as compared to H2O2 concentration from 0.5ppm to 5ppm resulted in
refrigerated samples. This is because the dehydoascorbic a tremendous increase in degradation rates which was
acid, the oxidized form of ascorbic acid was more stable recorded in Table 6. At 0.5ppm H2O2, the antioxidant
at lower temperatures. Thus, the vitamin C, in the form substances in orange juice which was flavonols reacted with
of dehydroascorbic acid for refrigerated orange juice was H2O2, thereby preventing the autoxidation of vitamin C.
well retained than non-refrigerated orange juice. The protective mechanism of flavanols was mainly due to
chelation of metal ions and action of antioxidant. Flavanols
Effect of Hydrogen Peroxide on The Stability of function as antioxidants by donating the hydrogen ions
Orange Juice to reactive free radicals which may otherwise cause the
Hydrogen peroxide, H2O2 is the primary chemical autoxidation of vitamin C.
for sterilization of plastic packaging material used in Figure 12:
aseptic system. Aseptic packaging technology is widely
used by fruit juice industry for the production of shelf-life
stable fruit juices. A Food and Drug Administration (FDA)
regulation currently limits the residual of H2O2 to 0.5ppm,
leached into distilled water, in finished food packages
which stated in Code of Federal Regulations, 2000.
However, during the sterilization of aseptic chambers or
packaging material with H2O2, some residues will still be
left on the packaging material or vapors generated during
drying may get trapped inside the package upon sealing.
These residues will then cause the degradation of vitamin
C.
An experiment was proposed by Ozkan, Kirca and
Cemeroglu to determine the rates of vitamin C degradation
in orange juice with or without addition of H2O2 at various
storage temperatures. In this experiment, the orange juice Ozkan, Kirca and Cemeroglu also studied the
sample was thawed at room temperature and sodium degradation of vitamin C in the absence of H2O2. In this
benzoate was added to prevent spoilage. The degradation case, the activation energy, Ea was taken into account to
studies were done at H2O2 with 0.5ppm concentration at determine the stability of vitamin C in orange juice. The
20anddeg;C, 30anddeg;C and 40anddeg;C respectively. temperature dependence of the degradation of vitamin
At regular time intervals, samples were removed from the C in orange juice was compared by calculating Ea and
water bath or incubator. Then, the predetermined amounts temperature quotients (Q10) at 20anddeg; to 40anddeg;C
of diluted sodium hydroxide solution were added rapidly to from the following equation:
the samples to halt the reaction between H2O2 and vitamin These results clearly indicate that the rate of
C. The samples were then rapidly cooled by plugging into vitamin C degradation in the presence of H2O2 was
an ice water bath and held at -30anddeg;C until analyzed for slower at 30anddeg;C to 40anddeg;C than 20anddeg;C
vitamin C content. Vitamin C concentration was measured to 30anddeg;C. This indicates that at 30anddeg;C to
by using HPLC method. Qzkan, Kirca and Cemeroglu 40anddeg;C, the least effect of temperature rise on vitamin
had modified the method by blending the orange juice C degradation. The results obtained for Ea shows that
sample with metaphosphoric acid. The sample was filtered higher Ea in the presence of H2O2. This means that higher
through a membrane filter and was analyzed using HPLC energy needed for the degradation of vitamin C. Therefore,
(Shimadzu brand).Vitamin C contents of orange juice were the reaction time is slower and the degradation of vitamin
plotted for various temperatures at 0.5ppm H2O2. C also slower. As conclusion, the effect of temperature on
From Figure 12, the results show that at higher the degradation rates of vitamin C in orange juice was more
temperature, the rate of vitamin C degradation also pronounced at higher H2O2 concentrations. Therefore,
increased. The addition of 0.5ppm H2O2 did not greatly greater vitamin C losses should be expected as residual
H2O2 concentration and storage temperature increase in 5.0 or below, dehydroascorbic acid was quite stable which
aseptically packaged fruit juices. decayed by less than 3% over 4 hours. This experiment
evaluated the effect of hydrogen ion concentration on
Effect pf Ph on The Stability of Vitamin C delactonization of dehyroascorbic acid over the range of
pH is a measure of acidity or basicity of a solution. pH 3.0 to pH 8.0. The possible influence of the presence
pH is one of the primary factor that would affects the of oxygen was done by equilibrating the reaction mixture
stability of vitamin C in orange juice. Hence, the pH value before and during the incubation with 100% oxygen or
of the matrix has an influence on the stability of vitamin C. with 100% nitrogen. The results indicated no change in the
According to FAO/WHO Expert Consultation on Human decay rate of dehydoascorbic acid was obvious with these
Vitamin and Mineral Requirements, Bangkok, Thailand, alterations of atmospheric conditions[16-18]. The rate of
1998, the vitamin C will decay if the pH higher than 4. dehydroascorbic acid hydrolysis markedly increases with
Vitamin C is unstable in neutral and alkaline increasing temperature but was unaffected by the presence
environments, therefore the higher the pH value and the of oxygen (Bode, Cunningham and Rose, 1990 Bode,
longer the exposure, the greater the loss of vitamin C. This M., Cunningham, L. and Rose, R. C. 1990. Spontaneous
is because the higher the pH value, the faster the oxidation Decay of Oxidized Ascorbic Acid (Dehydro-L-ascorbic
reaction of vitamin C and causes the degradation of acid) Evaluated by High-Pressure Liquid Chromatography.
vitamin C. Besides that, the increase in pH also related to Clinical Chemistry. Other researchers had proposed a
deterioration of fruit characteristic which in this literature method to determine the effect of pH on the degradation
review, orange juice is more concerned. Table 5 below of vitamin C in orange juice. The aim of their experiment
shows the pH value of the fruit juice with storage time. was by comparing the stability of vitamin C at different
Table 6: concentrations at lower pH value. An acidic sample was
Table 6: Ascorbic Acid Content (mg/100mL) at Different Storage prepared from orange juice with medium alcohol content
Times after Squeezing [11]. The original pH value of sample was later modified
by addition of concentrated phosphoric acid. After that,
NATURAAL ORANGE JUICE
different concentrations of vitamin C stock solutions were
Mean ±0 (% Retention) added and analysed for five weeks. The stability of vitamin
Time 1 2 C in different beverages. British Food Journal.
0 55.26 ± 0.22 5054 ± 0.15(100) The results showed that, for the experiment done
1 53.85 ± 0.14(97.45) ____ in original pH value of the orange juice which was pH 4.0,
3 52.40 ± 0.33(94.82) 50.36 ± 2.44(99.64) the reduction of the amount of vitamin C content decreased
5 50.76 ± 0.97(91.86) ____ with the increasing ascorbic acid concentration (p> 0.05),
6 50.00 ± 0.97(90.48) 50.33 ± 0.47(99.58) so the speed of decay was higher at lower concentrations.
24 48.20 ± 3.27(87.22) 50.22 ± 0.60(99.45)
In the case of orange juice, the highest standard deviation
of the repeated data was 3.14% and the lowest standard
In this the pH values of the orange juice were deviation was 1.48%. This indicates that the results are
higher at room temperature pH values keep increasing accurate and reproducible.
from week to week. This study concluded that, though pH When lower pH (pH 3.0) was used, the speed of
was significant for the stability of vitamin C, it was not the decay for orange juice grew with the growing vitamin C
sole factor in controlling the deterioration of vitamin C in concentration, and the highest value was at 50mg/L vitamin
orange juice with storage life. On the other hand, the loss of C concentration (p> 0.05). These results were tabulated in
vitamin C activity during oxidative degradation of vitamin Table 10. The highest standard deviation of the repeated
C occurs with the hydrolysis of the dehydroascorbic acid data was 3.42% and the lowest standard deviation was
lactone to yield 2,3-diketogulonic acid. This hydrolysis is 1.53%. By comparing the statistical data, it shows that the
favored by alkaline solution. Dehydroascorbic acid is most lower pH values increased the vitamin C content measured
stable at pH 5.5 but decrease in stability as pH increases at the end of fifth week.
which is more than pH 5.5. For example, half-time values According to Nagymate and Fodor, vitamin C
of dehydroasorbic acid hydrolysis at and deg;C were 100 had a significant decay independently from the storage
and 230 minutes at pH 7.2 and pH 6.6 respectively. At pH temperature when the pH value was more than 4.0.
(RQflex 10) which is a highly sensitive reflectrometry the product samples contain about 1.3 to 1.5 mg/L of HMF.
instrument. The combination of these two tests which are Based on the Commission Regulation, the average usage
the test strips and the reflectometer help to analyze the of HMF is 2.0mg/L and the maximum usage is 10.0mg/L
vitamin C content for a wide variety of samples in just for non-alcoholic products. HMF is a crystalline product
few seconds. Reflectometer uses a double optic system with a pleasant odor. HMF is formed in foods by thermal
which works in conjunction with a dual reaction zone on treatment during storage. High amount of HMF can cause
Reflectoquant Test Strips to allow for simultaneous double vitamin C loss, hence affecting the quality of the product.
measurements in one step. It is a portable test system Evaluation and optimization of non enzymatic browning
that is small, compact, and battery-operated for rapid, of “cajuina” during thermal treatment. Brazilian Journal of
quantitative analysis of various samples by evaluation of Chemical Engineering.
special test strips [18].
Vitamin C contents in each sample which is Determination of Vitamin C by Using Titration Method
determined using reflectometer do not have much Standardization of Sodium Hydroxide (NaOH) Solution
different compared to the label value. Tablets are labeled Using Potassium Acid Phthalate Solution (KHP)
according to their vitamin C content and not according to In a titration, it is critical to know the exact
their weight. The percentage of deviation of each sample concentration of NaOH in order to determine the
was calculated and the percentages of deviation obtained concentration of the solution being tested. KHP is a weak
for all the samples are less than 5.0%. This indicates acid and reacts with base in the following way:
that reflectometer can provide accurate and reproducible In the titration method, phenolphthalein was used
results. The difference between label value and analysis as an indicator which will be used to determine when
results could be caused by the interference substances the reaction reaches its endpoint. Endpoint is the point
such as the presence of binder. Binders are commonly which the amount of NaOH added equals the amount of
used when making conventional tablets. Most binders vitamin C. By knowing the strength and the volume of
are polymers which can increase the plastic deformation NaOH required to completely react with vitamin C, the
of the formulation. Binder can be used to prevent a rapid actual amount of vitamin C present can be calculated. pH
dissolution of the effervescent tablet such as Redoxon. meter also helps to determine the end point which is about
Examples of binders are such as methyl cellulose and pH8.50. As vitamin C is a weak acid, the pH of the end
gelatin which function to hold the ingredients together to point is detected by using phenolphthalein indicator with
form a tablet. For Redoxon, the analysis result is lower the transition range between pH8.0-9.8. Phenolphthalein
than the label value. This is due to the presence of foreign will changes from colorless to pink when all of the acid
substances such as zinc citrate which may influence the has been neutralized.
concentration of vitamin C. The samples were analyzed by the reflectometer.
The results show that both methods are in agreement with
Determination of Glucose Content and the quantities specified on the label. This indicates that
Hydroxymethylfurfural Content In Vitamin C Tablet the proposed method was applied successfully for the
The glucose content in each tablet of different determination of vitamin C in commercial pharmaceutical
brands is less than 1g/mL except for Flavette Vitamin products.
C. Flavette Vitamin C contains 22mg/L of glucose
content although this produc is labeled sugar free. Many Conclusion
manufactures use glucose, fructose or dextrose to sweeten Vitamin C is required for the optimal activity of
a tablet for commercial purpose even though the tablet several important biosynthetic enzymes and it is therefore
labeled ‘no sugar’. For hydroxymethylfurfural (HMF) essential for various metabolic pathways in the body.
test, 5-(Hydroxymethyl)furfural (5-hydroxymethyl-2- However, according to RDA for vitamin C, 75mg/day
furancarbaldehyde, HMF) reacts with a barbituric derivative and 90mg/day are required for normal women and men
and an aminophenazone derivative to form a red-violet respectively. This level is believed sufficient enough to
compound that is determined reflectrometrically. HMF prevent deficiency disease but not chronic disease. Owing
test was done to test the amount of undesirable product to this, vitamin C should be taken each day to prevent
such as 5-HMF in vitamin C tablet. From the results, all chronic disease and the effective doses are still remained