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Der Pharma Chemica, 2015, 7(8):46-49

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ISSN 0975-413X
CODEN (USA): PCHHAX

A comparitve study between the moist heat versus microwave method for the
determination of vitamin C content in amla
Sivaraj Anbarasu, Revathy Sundar, Jerrine Joseph* and Vanaja Kumar

Centre for Drug Discovery and Development, Sathyabama University, Chennai, India
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ABSTRACT

Amla/Indian gooseberry has a wide range of biological effects including antioxidant, antiviral, antimicrobial,
antitumor and antibacterial activities. The fruits of the plant have been used in Ayurveda as a potent rasayana
which is used to promote health and longevity. Amla is a rich source of polyphenols, minerals and vitamin C. This
study is based on the investigations carried out to assess the loss of vitamin C (ascorbic acid) in Amla during moist
heat and microwave oven method. The vitamin C content in moist heat and Microwave oven treated was noted to be
about 888 mg /100 g and 881 mg/100g respectively. The yield of vitamin C estimated by titration showed that the
yield enhanced from 983 to 1283mg to support the fact that the vitamin C levels have gradually increased with
increase in power in microwave oven extraction method when compared to the moist heat extraction method. Thus
the outcome is of translational value to the beverages and neutraceutical industry for shelf life improvement of the
Vitamin C based health drinks.

Keywords: Amla, ascorbic acid, neutraceutical, microwave and moist heat

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INTRODUCTION

Emblica officinalis, commonly known as amla/Indian gooseberry, belongs to the family of Euphorbiaceae. India
ranks first in the area of amla crop production in the world. It has wide range of biological effects including
antioxidant, antiviral, antimicrobial, antitumor and antibacterial activities. The fruits of the plant have been used in
Ayurveda as a potent rasayana which is used to promote health and enable increasing the lifespan by boosting
immune system, arresting the ageing process and revitalizing the body in debilitating conditions. Amla is a well
known rich source of polyphenols, minerals and vitamin C. The content of vitamin C is ranging from 200‐900 mg
per 100 g of the edible portion [1,2,3]. Vitamin C is regarded as the first line of natural antioxidant defense in
plasma and a powerful inhibitor of lipid peroxidation. It also regenerates the major antioxidant tocopherol (vitamin
E) in lipoproteins and cell membranes. Intracellular mechanisms exist which can regenerate vitamin C (ascorbate)
from its inactive metabolite dehydroascorbate by reduced glutathione [4]. The active ingredient of Amla comprises
of gallic acid or ellagic acid structures attached to vitamin C. The leaves and bark are rich in tannin. The pH value of
10% w/v of aqueous solution is acidic [5,6]. TSS, pH, reducing sugars, total sugars and browning of the preserves
increases during storage irrespective of different methods employed for preparation of the preserves. Moisture,
ascorbic acid, tannin and titratable acidity of the preserves decreased during storage. The preserves at sugar syrup of
70°Bricks was found most effective in retention of ascorbic acid and tannins and showed low microbial load [7].

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The amla fruit extract is also a good source of polyphenols, flavones, tannins and other bioactive substances. These
substances are also strong antioxidants and might contribute to improved health benefits. Thus it is important to
characterize different types of medicinal plants for their antioxidant and antimicrobial property. E. officinalis alone
or in combination with other herbs has been useful in the treatment of cold, warts, skin afflictions, influenza,
anemia, diabetes, lung conditions and elevated cholesterol as an immune restorative in cancer conditions. It is one of
the best natural anti-ageing remedies used in acne and other skin problems and also effective in the treatment of
acidity and peptic ulcers. Regular use of amla improves immunity, fights cancers and rejuvenates the body. It fights
chronic diseases like hypertension, high cholesterol, Diabetes, AIDS, influenza, chronic infections like cough and
cold, fatigue and inflammatory conditions. The amla seeds and leaves are used in asthma, bronchitis, biliousness,
conjunctivitis, inflammation, dyspepsia and dysentery. Liquor fermented from amla fruit is good for indigestion,
anemia, jaundice, heart complaints and for promoting urination [8]. Ayurveda describes amla as one of the best cure
for diabetes, bleeding disorders, strength and stamina promoter. Clinical studies on patients with pulmonary
tuberculosis showed that vitamins of E. officinalis was better assimilated than synthetic vitamin C [9]. The amla
extracts showed significant protection to DNA against oxidative damage as found by migration of DNA in agarose
gel. Amla inhibits radiation induced lipid peroxidation (LPO) in microsomes and superoxide dismutase (SOD) in
mitochondria. As amla fruits are perishable in nature, its shelf life during storage is very limited. The contents of
ascorbic acid decreases with increase in storage period. Its decay loss can be minimized by storing in regulated
conditions. It was found that decay loss was minimum in modified storage condition whereas it was maximum in
zero energy chamber. The chemical changes occur during processing and storage after 135 days. It was also
observed that total soluble solid content, the level of vitamin C, tannin, etc., are decreased as in fresh fruits but they
are rich in calcium, sugar and further increased acceptability after storage of upto 135 days [10,11]. A mature amla
can tolerate freezing as well as high temperature of 46⁰C. Amla being water soluble, it may scavenge the free
radicals responsible to initiate lipid peroxidation. However, ascorbic acid alone is not responsible for antioxidant
activity. Other polyphenolic compounds capable of scavenging oxidizing radicals also contribute to it. Ascorbic acid
in amla itself does not provide high protection even at elevated concentration. Therefore, ascorbic acid and other
polyphenols present in natural formulation of amla have higher antioxidant activity than equivalent amount of
ascorbic acid present in synthetic form. Present study is unique in a way that it investigates to improve the yield of
vitamin C (ascorbic acid) from amla by comparing the methodologies of moist heat and microwave oven methods.

MATERIALS AND METHODS

a. Titration method
The vitamin C (ascorbic acid) content in moist heat and microwave oven heat treated amla were estimated by
titration using iodine solution. Briefly, 10 ml of filtrate (sample) was taken in 100ml conical flask and 10 drops of
1% starch indicator solution was added. Iodine solution was added using burette into the conical flask containing
the sample. The end point was noted by the appearance of blue-black complex.

b. Preparation of fruit pulp

Amla fruits were cut into small pieces and seeds were removed. About 200 grams of pulp alone was weighed. It
was crushed by using mortar and pestle by slowly adding 200 ml of sterile distilled water. Two sets of preparations
were made separately as mentioned above for moist heat (MH) and microwave oven (MO) methods.

c. Moist Heat method (MH)

About 50 ml of MH sample was filtered through Whatman No.1 filter paper and filtrate was collected in separate
conical flask (MHC). Remaining 50 ml of MH sample was subjected to autoclave at 1210C for 15 minutes and then
filtered as mentioned above (MHF). Both the samples were titrated for vitamin C content in duplicate as per the
titration protocol mentioned above.

d. Microwave Oven Method (MO)

The MO sample was divided into four parts each weighing 30 grams and equal volume of sterile distilled water was
added to all of them. The first part was kept in microwave oven (IFB 20SC2) at power 60 (60%) for 240 seconds
(MO60). The second part was subjected to heat at power 80 (80%) for 140 seconds (MO80), and third part was
treated at power 100 (100%) for 75 seconds (MO100). The final part of sample was not kept at microwave oven
(MOC). The time noted in all experiments was based on the boiling point of the particular sample. The raise in
above time durations causes boiling and leads to spillage and wastage of the sample. The samples were filtered using

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Whatman No.1 filter paper and collected the filtrate in separate conical flask. Then the samples were titrated for
vitamin C content in duplicate as per the titration protocol mentioned.

RESULTS AND DISCUSSION

The vitamin C content in MHF and MHC was noted to be about 888 mg /100 g and 881 mg/100g respectively
(Table 1). This shows that the effect of heat does not have any noteworthy increase in Vitamin C content as also
corroborated with a report that the ascorbic acid content of fresh amla fruit itself exhibited the value of 950mg/100
gm [12]. The moist heat does not have a great impact on the vitamin C content in the amla pulp. However, vitamin C
in amla was lost during cooking more in open pan than pressure cooking [13]. In our study, when compared to the
moist heat exposure, the microwave oven treated samples yielded higher quantity of vitamin C and the values have
gradually increased from 983mg/100g to 1283 mg/100g with increasing power (Table 2 & Fig.1). Vitamin C content
of amla increases in the sun dried samples (for example 100 grams of fresh amla gives out 600 mg of Vitamin C), its
content increasing from 1500 to 1600 mg [14]. The moist heat treated filtrate sample was turned into brownish
colour after autoclave, whereas no colour change was observed in sample during the microwave oven treatment.
Browning of amla occurs in heating at higher temperature compared to lower temperature [15]. This shows that the
colour changes attributed to the decrease in vitamin C content. Hence this method will not be ideal for packaging
and storing of neutraceuticals where the shelf life of the Vitamin C content has to be maintained over a period of
several months.
Table1: Vitamin C content in moist heat treated samples

Sample ID Temperature Time in minutes Average Vitamin C per 100g

MHC NIL NIL 881 mg

MHF 121oC 15 888 mg

Table2: Vitamin C content in microwave oven treated samples

Sample ID Power Time in seconds Average Vitamin C per 100g

MOC NIL NIL 849 mg
MO60 60 (60%) 240 983 mg
MO80 80 (80%) 140 1275 mg
MO100 100 (100%) 75 1283 mg

Fig1: Increasing vitamin C content during microwave oven treatment

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CONCLUSION

In conclusion, the results are encouraging to support the fact the vitamin C levels have gradually increased when
heated in microwave oven corresponding to the increase in power, compared to the moist heat extraction. Hence the
microwave method is more suited to extract the vitamin C. The outcome could be of translational value to the
beverages and neutraceutical industry where Vitamin C levels are very crucial and should be maintained at high
levels to bring the required impact in the individual as immune booster through their products.

Acknowledgement
The authors are thankful to Chancellor and Directors of Sathyabama University, providing financial support and the
lab facility for the work.

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