BIOLOGIA (LAHORE) PKISSN 0006 – 3096 (Print)

December, 2021, 67 (II), 134-140 ISSN 2313 – 206X (On Line)

UV/Vis Spectrophotometric Quantification of Vitamin C in Leafy Vegetables

Under Different Storage Conditions
MUHAMMAD WAHEED MUSHTAQ1*, NAUMAN JAMIL KHAN1, NAVEED ASLAM DOGAR1, AISHA
HAMEED2, ABDUL HAMEED2, MALIK ADIL2, HAROON IFTHIKHAR3 & SAIRA ISHAQ4
1
Chemistry Department, Govt. Graduate College of Science, Wahdat Road, Lahore, Pakistan.
2Botany Department, Govt. Graduate College of Science, Wahdat Road, Lahore, Pakistan.
3
Institute of Industrial Biotechnology, GC University, Lahore Pakistan
4Govt. Postgraduate Islamia College, Cooper Road, Lahore.

ARTICLE INFORMAION ABSTRACT

Received: 09-13-2021 Leafy vegetables (LVs) have health promoting phytochemicals and are
Received in revised form: rich source of many nutrients such as Vitamin C (Vit. C). The aim of this
02-05-2022 study was to design an easy, accurate, sensitive and fast method to
Accepted: 23-05-2022 determine concentration of vitamin C (ascorbic acid) from leafy
vegetables. So, concentration of Vit. C was measured
*Corresponding Author: spectrophotometrically by making a red color complex between 1, 10
Phenanthroline and Vit. C in the presence of ferric chloride (FeCl3). This
colored product has maximum absorption at lambda max (λmax= 430
Muhammad Waheed nm). Ascorbic acid was estimated from leafy vegetables placed at three
Mushtaq: different conditions such as firstly, from fresh vegetables as available in
waheedjaami@[Link] market, secondly stored for 2 days at room temperature (25±2 ºC) and
thirdly, stored in refrigerator (4 ºC) for 2 days. It was observed that
vegetables stored at room temperature was significantly affected by loss
of Vit. C as compared to refrigerated LVs. The average % age loss of
ascorbic acid observed in refrigerated and room temperature stored
vegetables was 34.18 % and 40.9 % respectively, as compared to fresh
one. The highest concentration of vitamin C in fresh LVs was shown by
spinach (268.7 mg/L) and least by lettuce (98.03 mg/L). This study shows
that fresh vegetables are good source of vitamin C as compared to stored
vegetables.
Original Research Article Keywords: Leafy vegetables, Vitamin C, Spectrophotometer

INTRODUCTION body functions, including formation of collagen,

absorption of iron, the proper functioning of the
Vitamins are significant nutrients that immune system, wound healing, and the
cannot be prepared by our body so they must be maintenance of cartilage, bones, and teeth. Many
supplied through our diet for biochemical and healthy benefits are of the Vit. C such as necessary
physiological processes. Casimir Funk originally for the growth, development and repairing of all
coined the term “vitamine” in 1912 (Semba et al., body tissues.
2012). Vitamins are further classified into two
categories i.e fat soluble and water soluble
vitamins. Vitamins A, D, E and K are fat soluble
whereas vitamin C and vitamin B complex are
water soluble vitamins. The chemical name of
vitamin C is ascorbic acid. It is L- enantiomer of
ascorbic acid. It is a strong reducing agent and
antioxidant. Many factors effects Vit. C stability Fig 1: Structure of vitamin C (Ascorbic acid)
such as temperature, oxygen and light (Dave &
Patil, 2017). The structural formula of Vit.C is It also helps in preventing pathological
shown in Fig 1 (Elgailani et al., 2017). disease scurvy, provides relief from common cold
Due to its biological importance Vit. C is and is involved in wound healing (Iqbal et al., 2004).
very important and has great significance in many It can be used in treatment of many chronic type of

Author Contribution: M. W.M. & N.A.D. & S. I., Helped in writing and experimentation; N. J. K., Designing and supervision of project; A.
H., Performed lab work; A. H. & M. A., Provided chemicals and instruments; H.I., Helped to check plagiarism.
VOL. 67 (II) SPECTROPHOTOMETRIC DETERMINATION OF VITAMIN C 135

cancer such as colon, oral and prostate cancer MATERIALS AND METHODS
(Levine, 1986; Block, 1991). It aids in the formation
of collagen and fabulous healthy skin, cartilage, Reagents and Chemicals
ligaments, corneas etc. Because of antioxidant
property, it act as free radicals scavenger (Mazloum Analytical grade level reagents and
Ardakani et al., 2018). HIV- patients are often chemicals are used in this procedure. Ferric
prescribed Vit. C to protect their defending system Chloride (FeCl3, BDH 99%), ascorbic acid, 1-10
and plays its important role in defending chronic Phenanthroline (BDH 99%), Hydrochloric acid
diseases like tuberculosis disease and AIDS (HCl). To determine the amount of ascorbic acid,
(Makinde et al., 2017). Detoxifing ability of toxic UV/visible spectrophotometer (Germany, UV-3000)
substances of liver can be enhanced by Vit. C was used with range of 300nm-800nm.
(Matei & Birghila, 2001; Mushtaq et al., 2022).
Fruits are primary and significant sources of Experimental Procedure
Vit. C such as citrus fruits, peaches, red grapes, Preparation of Samples
pineapple, pomelo, grape fruit contain vitamin C in
great range (Matei & Birghila, 2001). A significant Selected LVs such as spinach, coriander,
amount of Vit. C is also found in green leafy mint, lettuce, fenugreek (Methi) and cabbage were
vegetables such as in Parsley, coriander, spinach, purchased from market as fresh vegetables. These
fenugreek etc. (Matei et al., 2004). vegetables were washed three times with tap water
Shelf life of vegetables can be increased by to remove dirt. 50 g of each vegetable were ground
refrigeration which helps in retention of Vit. C in mortar and pestle separately to get the extract.
content. Vit. C components depend upon different The obtained extract was sieved to get pure filtrate
conditions such as weather, ripeness, processing, of each vegetable in the form of juice. Finally, 10 ml
storage and preparation of each type of food. of each vegetable filtrate was added in three test
Cutting, cooking, washing, thermic processes and tubes separately, and labelled them as extract 1, 2
exposure to atmospheric oxygen cause a significant and 3 for fresh, room temperature (25 ºC) stored
loss of Vit. C content. Vit.C concentration is and refrigerator stored (4 ºC) vegetables
gradually varied by season, transportation to respectively. The quantity of Vit. C in fresh
market, time of storage, cooking practices and shelf vegetables was measured immediately but for
life (Vina & Chaves, 2006). stored vegetables, the amount was measured after
Various classical and instrumental methods two days of storage.
such as Titration, potentiometry, amperiometry, To measure ascorbic acid concentration, 5
fluorimetry, voltammetry, HPLC, TLC etc. can be mL of each vegetable extract was taken separately
applied to quantify the concentration of ascorbic in a test tube. Then 1 mL of ferric chloride (FeCl3)
acid. Spectrophotometric methods for determination and 4 to 5 drops of 1,10 Phenanthroline solution
of ascorbic acid depend upon oxidation-reduction were added in each test tubes. After continuous
reaction because of its reducing property. UV stirring for two minutes, a dark red colour solution
radiations can be easily absorb by Vit. C that’s why was obtained whose absorbance was noted on
spectrophotometry is one of the most frequently spectrophotometer at 430 nm.
used simple methods and is considered one of the
best method with less chances of error (Silva, 2005; RESULTS
Timothy 2021). Calibration Curve
The purpose of present study was to
measure spectrophotometrically quantity of Vit. C in To measure the concentration of ascorbic
selected LVs available in Shalimar Link road, acid in samples, a standard calibration curve was
Lahore (Pakistan). The LVs included in this study drawn for different dilutions (20-300 mg/L) of
were Spinacia oleraceae , Coriandrum sativum, ascorbic acid. The absorbance of dilutions was
Mentha spicata, Trigonella foenum- graecum , measured at λmax of 430 nm. The standard curve
Brassica oleraceae, Lactuca sativa. The amount was plotted between concentration and the
of Vitamin C from these LVs, was estimated in three corresponding absorbance as shown in Figure 3.1.
modes such as fresh available in market, stored at The straight line indicated that absorbance is
room temperature (25±2 ºC) for 2 days and stored directly related to ascorbic acid concentration.
in refrigerator (4 ºC). With the help of calibration curve, the
measured concentration of ascorbic acid in fresh,
room temperature stored (25 °C, 2 days) and
refrigerated stored (4 °C, 2 days) vegetables are
 136 M.W. MUSHTAQ ET AL BIOLOGIA LAHORE

shown in Fig 3.2 – 3.4. Fig 3.5.

The graph plotted between % loss of Vit. C
between fresh and vegetables stored for 2 days
under room temperature is shown in Fig 3.6.

Fig 3.1: Calibration curve for ascorbic acid

Fig 3.4 (a-b): Room temperature stored vegetables

extract showing ascorbic acid concentration test, RT1
(cabbage), RT2(fenugreek), RT3 (mint), RT4 (spinach),
RT5 (coriander), RT6(lettuce)

Fig 3.2 (a-b): Fresh vegetables extract showing ascorbic

acid concentration F1(spinach), F2(fenugreek), F3(mint),
F4(coriander), F5(cabbage), F6(lettuce)

Fig 3.5: Quantity of % age loss (Vit. C) between market

fresh and refrigerated vegetables

Fig 3.3 (a-b): Refrigerated vegetables extract showing

ascorbic acid concentration;
R1 (spinach), R2 (fenugreek), R3 (mint), R4 (coriander),
R5 (cabbage), R6 (lettuce)

The graph plotted between % age loss Fig 3.6: Quantity % age loss of Vit. C between market
between market fresh vegetables and refrigerated fresh and stored vegetables for 2 days at room
vegetables at 4ºC storage for 2 days is shown in temperature
VOL. 67 (II) SPECTROPHOTOMETRIC DETERMINATION OF VITAMIN C 137

DISCUSSION degrades. (Souzan & El-Aal, 2000).

Results indicated that the concentration of
Vit. C is usually quantified by various Vit. C in market fresh vegetables was 98.035 mg/L
methods such as HPLC, TLC, LC, potentiometric, in lettuce, 268.7 mg/L in spinach, 230.24 mg/L in
voltametric, amperometric and titrimetric method. mint, 226.619 mg/L in coriander, 249.45 mg/L in
But there are some limitations with such methods fenugreek and 172.19 mg/L in cabbage. The order
such as in titrimetric method, food colour interferes of concentration of Vit. C in market fresh vegetables
with the concentration of Vit. C. Chromatographic was;
method is very costly. So the best method for the Spinach > Fenugreek > Mint > Coriander
quantification is spectrophotometric method. It is > Cabbage > Lettuce
more accurate and reliable. As titration and
chromatographic methods take more time for to the The spectrophotometric quantification
detection of Vit. C, we choose spectrophotometric indicated that the high concentration of Vit. C was
method. UV/Vis Spectrophotometry is one of the found in spinach (268.7 mg/L) and the least in
most unique and frequently used simple method lettuce (98.035 mg/L). The concentration of Vit. C
(Grosso et al., 2013; Loria et al., 2000; Moeslinger in refrigerated vegetables that are stored
et al., 1995). Spectrophotometric method described under temperature 4ºC for 2 days are 254.5
here is based on the principle of reduction of Ferric mg/L in spinach, 192.548 mg/L in fenugreek,
to Ferrous ions and to form a complex called 111.92 mg/L in mint, 176mg/L in coriander, 169.71
Ferroin by using ascorbic acid in presence of 1,10- mg/L in cabbage and 89.90 mg/L in lettuce. The
phenanthroline. Absorbance of the resulting Fe(II)- concentration of Vit. C in vegetables that are
phenanthroline red complex (Ferroin) was stored at room temperature for 2 days was
measured at 430 nm. measured as 164.7 mg/L in spinach , 181.04 mg/L
The concentration of Vit. C detected here in in fenugreek, 69.982 mg/L in mint, 161.66 mg/L in
fresh vegetables is higher as compared to coriander, 77.23 mg/L in cabbage and 78.03 mg/L
refrigerated vegetables and those which are stored in lettuce.
at room temperature. A loss of Vit. C is observed in Higher concentration is seen in fresh
refrigerated vegetables and stored vegetables for 2 vegetables as compared to refrigerated and
days. vegetables stored at room temperature for 2 days.
Vit. C is a water soluble compound which A significant loss of Vit. C was observed in
can easily leach into water and then degraded by refrigerated and stored vegetables. According to
heat. Vit. C is unstable molecule which can easily Von Hahn & Gorbing study, the % age loss in
be denatured by heat. It is very sensitive to beans, cauliflower, lettuce, parsley, peas and
oxidation and can be depleted easily. Refrigeration spinach was 33%, 8%, 36%, 13%, 10% and 32%
reduces the rate of respiration of vegetables and respectively when stored at 4 ºC for the duration of
that’s why refrigeration is often recommended for two days. Whereas the %age loss of Vit. C was
increased shelf life of vegetables (Khan et al., 2006; further increased by increasing temperature at 20
Wonsawat, 2014; Rickman et al., 2007; Murcia et ºC (Von Hahn & Gorbing, 1933). In the present
al., 2000; Bessey & King, 1933; Kifer & Munsell, study, the percentage losses determined for
1932; Wasson,1931; Langley et al., 1933). refrigerated vegetables were 5.28% (spinach),
Leafy vegetables wrapped in polythene 22.81% (fenugreek), 51.39% (coriander), 22.33%
was very effective in reducing weight and moisture (coriander), 1.44% (cabbage), 8.3% (lettuce) and
and helpful in retention of Vit. C content (Podsedek, for stored vegetables (for 2 days, 25 ºC) were
2007). Quality of fertilizer also effects the Vit. C 38.70% (spinach), 27.42% (fenugreek), 69.60%
content of the vegetable (Vina & Chaves, 2006). (mint), 28.66% (coriander), 55.148% (cabbage) and
Factors which effect Vit. C content are oxygen, 20.41% (lettuce).
temperature and light because Vit. C is easily Overall, the decrease in concentration of
oxidized with the exposure with oxygen. Vit. C was noted for refrigerated vegetables. The
Temperature also affects as it is water soluble and highest percentage loss was shown by refrigerated
dehydrated by heat easily. Some vegetables lose samples of Mentha spicata (mint). The average %
half of their vitamin C content when allowed to age loss observed in between fresh and
stand at room temperatures for 3 days, and their refrigerated LVs is 37.18 % age whether the % age
rate of degradation is determined lower when loss observed between fresh and vegetables stored
allowed to stand under refrigeration (Mushtaq et al., for 2 days at room temperature is 40.9 % age. This
2022). Depletion of Vit.C occurs gradually after average % age loss indicates that highest loss is
harvest and with the passage of time it steadily seen under room temperature storage as it is very
 138 M.W. MUSHTAQ ET AL BIOLOGIA LAHORE

labile to the action of heat heat and oxygen and Elgailani, I. E. H., Gad-Elkareem, M. A., Noh, E. A.,
therefore Vit. C activity slowly decreases during its Adam, O. E., & Alghamdi, A. M. (2017).
storage. As described earlier that we should Comparison of two methods for the
consume fresh vegetables rather than the stored determination of vitamin C (ascorbic acid) in
vegetables at 4 ºC in refrigerator or at normal room some fruits. American Journal of Chemistry,
temperature stored for 2 days. Market fresh 2(1), 1-7.
vegetables found to be the greatest source of Vit. Fediuk, K. (2000). Vitamin C in the Inuit diet: past
C. Refrigerated vegetables were found to lose less and present (Doctoral dissertation, McGill
% age loss as compared to vegetables stored at University).1-137.
room temperature for 2 days (Souzan & El-Aal, Fenoll, J., Martínez, A., Hellín, P., & Flores, P.
2000). This study shows that fresh vegetables are (2011). Simultaneous determination of
good source of Vit. C as compared to stored ascorbic and dehydroascorbic acids in
vegetables. vegetables and fruits by liquid
chromatography with tandem-mass
CONCLUSION spectrometry. Food chemistry, 127(1), 340-
344.
It can be easily concluded from the present Figueroa-Mendez, R., & Rivas-Arancibia, S. (2015).
work that performed UV/Vis Spectrophotometric Vitamin C in health and disease: its role in
method for determination of Vit. C is an easy, the metabolism of cells and redox state in
simple, accurate, precise, selective, safe and more the brain. Frontiers in physiology, 6, 397.
reliable. All the reagents used were cheap and Grosso, G., Bei, R., Mistretta, A., Marventano, S.,
readily available. So, by using this procedure Calabrese, G., Masuelli, L., & Gazzolo, D.
quantity of ascorbic acid can be measured in (2013). Effects of vitamin C on health: a
different samples quite rapidly. A significant loss in review of evidence. Front Biosci (Landmark
ascorbic acid concentration was found for Ed), 18, 1017-1029.
refrigerated vegetables. It can be concluded that we Hacisevki, A. (2009). An overview of ascorbic acid
should give preference for the consumption of fresh biochemistry. Ankara University Faculty of
LVs as Vit. C source over refrigerated vegetables at Pharmacy Journal, 38(3), 233-255.
4ºC as well as stored vegetables at room Hoffer, A. (1989). The Discovery of Vitamin C.
temperature. Albert Szent-Gyorgyi, MD (Doctoral
dissertation, Ph.D. 1893-1986. “to see what
REFERENCES everyone else has seen, but think what no
one else has thought”. Journal of
Alberg, A. J. (2002). The influence of cigarette Orthomolecular Medicine, 4(1), 24-26.
smoking on circulating concentrations of IOM (2000). Ascorbic acid. In: Dietary Reference
antioxidant micronutrients. Toxicology, Intakes for Ascorbic acid, Vitamin E,
180(2), 121-137. Selenium, and Carotenoids. Food and
Bessey, O. A., & King, C. G. (1933). The Nutrition Board, Institute of Medicine.
distribution of vitamin C in plant and animal National Academy Press, Washington DC;
tissues, and its determination. Journal of chapter 5, 1-506.
Biological Chemistry, 103, 687-698. Iqbal, K., Khan, A., & Khattak, M. M. A. K. (2004).
Block, G. (1991). Vitamin C and cancer prevention: Biological significance of ascorbic acid
the epidemiologic evidence. The American (vitamin C) in human health-a review.
journal of clinical nutrition, 53(1), 270-282. Pakistan Journal of Nutrition, 3(1), 5-13.
Bohndiek, S. E., Kettunen, M. I., Hu, D. E., Khan, M. R., Rahman, M. M., Islam, M. S., &
Kennedy, B. W., Boren, J., Gallagher, F. A., Begum, S. A. (2006). A simple UV-
& Brindle, K. M. (2011). Hyperpolarized [1- spectrophotometric method for the
13C]-ascorbic and dehydroascorbic acid: determination of vitamin C content in various
vitamin C as a probe for imaging redox fruits and vegetables at Sylhet area in
status in vivo. Journal of the American Bangladesh. Journal of biological sciences,
Chemical Society, 133(30), 11795-11801. 6(2), 388392.
Dave, K., & Patil, R. (2017). Biological Importance Kifer, H. B., & Munsell, H. E. (1932). Vitamin
of Ascorbic Acid (Vitamin C) in Human Content of Three Varieties of Spinach.
Health-A Classic Review. International Journal of Agricultural Research, 44, 767-
Journal For Research In Biology & 71.
Pharmacy. 3(7), 01-08. Langley, D. D., Richardson, J. E., & Andes, E. J.
VOL. 67 (II) SPECTROPHOTOMETRIC DETERMINATION OF VITAMIN C 139

(1933). The effect of storage and canning broccoli. Journal of the Science of Food and
upon the vitamin content of carrots. The Agriculture, 80(13), 1882- 1886.
effect of storage and canning upon the Mushtaq, M.W., S. Tariq, A. Hameed, S. Bashir, S.
vitamin content of carrots, 276, 1-32, Bibi, N.A. Dogar, A. Batool and S. Ishaq.
Levine, M. (1986). New concepts in the biology and (2022). Spectrophotometric determination of
biochemistry of ascorbic acid. New England vitamin C in underground vegetables and
Journal of Medicine, 314(14), 892-902. kinetic modelling to probe the effect of
Loria, C. M., Klag, M. J., Caulfield, L. E., & Whelton, temperature and pH on degradation of
P. K. (2000). Vitamin C status and mortality vitamin C. Pak. J. Bot., 54(5): 1-8.
in US adults. The American journal of Pathy, K. (2018). Process for Preparation of Vitamin
clinical nutrition, 72(1), 139-145. C and Method for Determination of Vitamin
Majidi, M. I. H. A., & AlQubury, H. Y. (2016). C in Tablets. Sci Fed Journal of Chemical
Determination of vitamin C (ascorbic acid) Research, 2(1), 1-14.
contents in various fruit and vegetable by Podsedek, A. (2007). Natural antioxidants and
UV-spectrophotometry and titration antioxidant capacity of Brassica vegetables:
methods. Journal of Chemical and A review. LWT-Food Science and
Pharmaceutical Sciences, 9(4), 2972-2974. Technology, 40(1), 1-11.
Makinde, O., Rotimi, K., Ikumawoyi, V., Adeyemo, Rickman, J. C., Barrett, D. M., & Bruhn, C. M.
T., & Olayemi, S. (2017). Effect of vitamin A (2007). Nutritional comparison of fresh,
and vitamin C supplementation on oxidative frozen and canned fruits and vegetables.
stress in HIV and HIV-TB coinfection at Part 1. Vitamins C and B and phenolic
Lagos University Teaching Hospital (LUTH) compounds. Journal of the Science of Food
Nigeria. African health sciences, 17(2), 308- and Agriculture, 87(6), 930-944.
314. Rivas, C. I., Zuniga, F. A., Salas-Burgos, A.,
Mandl, J., Szarka, A., & Banhegyi, G. (2009). Mardones, L., Ormazabal, V., & Vera, J. C.
Vitamin C: update on physiology and (2008). Vitamin C transporters. Journal of
pharmacology. British journal of physiology and biochemistry, 64(4), 357375.
pharmacology, 157(7), 1097-1110. Semba, R. D. (2012). The discovery of the vitamins.
Matei, N., & Birghila, S. (2001). Determination of Int J Vitam Nutr Res, 82(5), 310-315.
Vitamin C from fruit juices and Silva, F. O. (2005). Total ascorbic acid
pharmaceuticals. Ann. Univ. Ovidius, 12, 47- determination in fresh squeezed orange
49. juice by gas chromatography. Food Control,
Matei, N., Birghila, S., Dobrinas, S., & Capota, P. 16(1), 55-58.
(2004). Determination of C vitamin and Souzan, S. L., & El-Aal, H. A. (2007). Minerals
some essential trace. Acta Chim Slov, 51, profile-shelf life extension and nutritive value
169-175. of fresh green leafy vegetables consumed in
Mazloum-Ardakani, M., Abolhasani-Soorki, M., Egypt. In 8th African Crop Science Society
Khoshroo, A., Sabaghian, F., & Mirjalili, B. Conference, El-Minia, Egypt, 27-31 October
F. (2018). Simultaneous Determination of 2007. African Crop Science Society.
Ascorbic Acid, Uric Acid and Tryptophan by Teucher, Olivares, & Cori. (2004). Enhancers of
Novel Carbon Nanotube Paste Electrode. iron absorption: ascorbic acid and other
Iranian journal of pharmaceutical research: organic acids. International journal for
17(3), 851. vitamin and nutrition research, 74(6), 403-
Moeslinger, T., Brunner, M., Volf, I., & 419.
Spieckermann,P.G.(1995).Spectrophotomet Timothy, M. M. (2021) "Development and Validation
ric determination of ascorbic acid and of a UV/vis Spectrometric Method for
dehydroascorbic acid. Clinical chemistry, Determination of Cefixim Trihydrate in Raw
41(8), 1177-1181. Materials (Pure) and Tablet
Morabia, A., & Wynder, E. L. (1990). Dietary habits Forms." International Journal of Pharmacy
of smokers, people who never smoked, and and Chemistry 7(2): 31.
exsmokers. The American journal of clinical Vina, S. Z., & Chaves, A. R. (2006). Antioxidant
nutrition, 52(5), 933-937. responses in minimally processed celery
Murcia, M. A., López‐Ayerra, B., Martinez‐Tome, during refrigerated storage. Food Chemistry,
M., Vera, A. M., & García Carmona , F. 94(1), 68-74.
(2000). Evolution of ascorbic acid and Vina, S. Z., & Chaves, A. R. (2006). Antioxidant
peroxidase during industrial processing of responses in minimally processed celery
 140 M.W. MUSHTAQ ET AL BIOLOGIA LAHORE

during refrigerated storage. Food Chemistry,

94(1), 68-74.
Von Hahn, F. V., & Görbing, J. (1933). Influence of
fertilizers on the vitamin C content of
spinach. Z. Untersuch. Lebensm, 65, 601-
616.
Wasson, G. E. (1931). The vitamin C content of
fresh and canned spinach, Swiss chard,
asparagus and carrots. 4(1), 1-29.
Wonsawat, W. (2014). Determination of Vitamin C
(Ascorbic Acid) in Orange Juices Product.
World Academy of Science, Engineering
and Technology, International Journal of
Biological, Biomolecular, Agricultural, Food
and Biotechnological Engineering, 8(6), 623-
625.