A

PROJECT REPORT
ON
Identification Of Essential Vitamins In
Balance Diet

MORNING STAR
ST. ANSELM" SCHOOL
JAIPUR
SUBMITTED TO: SUBMITTED BY:

Ms. Anjali Peter Aditya Sharma

Class XI A

ACADEMIC YEAR 2022-23

DEPARTMENT: CHEMISTRY
 Certification
THIS is to certify that "Aditya Sharma" student
of class 11A has successfully completed his
physics project under the guidance of "Ms.
Anjali Peter"

Ms. Anjali Peter Fr. Victor Raj

(Teacher) (Principal)
 Acknowledgement

I would like to express my special thanks of

gratitude to my subject teacher Ms. Anjali
Peter to give her guidance to make the
successful completion of this project.
I also want to give special thanks to our
principal Father Victor Raj who gave me this
golden opportunity to do this wonderful project
on the topic “Analysis of vegetable and fruit
juices”, so that I will get to know about
detailed information for the same.
Secondly I would like to thank my parents and
classmates who helped me to complete this
project within the given time frame.
Abstract
Aim is to analyze some fruits & vegetables juice for
the contents present in them. Fruits and vegetable
are always a part of balanced diet. That means
fruits and vegetables provide our body the essential
nutrients, i.e. Carbohydrates, proteins, vitamins and
minerals.
Again their presence in these is being indicated by
some of our general observations, like -freshly cut
apples become reddish black after some time.
Explanation for it is that iron present in apple gets
oxidized to iron oxide. So, we can conclude that
fruits and vegetables contain complex organic
compounds, for e.g., anthocin, chlorophyll, esters
(flavoring compounds), carbohydrates, vitamins and
can be tested in any fruits or vegetables by
extracting out its juice and then subtracting it to
various tests which are for detection of different
classes of organic compounds. Detection of
minerals in vegetables or fruits means detection of
elements other than carbon, hydrogen and oxygen.
Table of Contents
List of Abbreviations and / or Glossaries.............................................. I
Dedication...................................................................................................................... II
Acknowledgment....................................................................................................... III
Table of Contents..........................................................................................................
IV
1. Introduction................................................................................................................ 1
2. Difinition..................................................................................................................... 2
2.1 Fat soluble vitamins............................................................................................ 3
2.1.3 Vitamin E........................................................................................................ 3
2.1.2 Vitamin D........................................................................................................ 4
2.1.3 VitaminA........................................................................................................ 6
2.1.4 Vitamin K....................................................................................................... 7
2.2 water soluble vitamin ............................................................................................. 8
2.2.1 Thiamin .......................................................................................................... 8
2.2.2 Riboflavin ...................................................................................................... 9
2.2.3 Ascorbic acid ................................................................................................. 10
3. Food source of vitamins................................................................... 11
4. Vitamin analysis in foods...................................................................................... 12
5. Methods used for analysis of vitamins: .......................................... 12
5.1. High-performance liquid chromatography……………………............ 13
5.2. Gas chromatography ...........................................................................................
14
5.3. liquid chromatograph-ass spectrometry ......................................15
6. Importance of Analysis.................................................................... 15

References .....................................................................................................................
16
Introduction
1. Introduction The vitamins are a disparate group of compounds; they have little in common
either chemically or in their metabolic functions. Nutritionally, they form a cohesive groupof
organic compounds that are required in the diet in small amounts (micrograms or milligrams per
day) for the maintenance of normal health and metabolic integrity. They are thus differentiated
from the essential minerals and trace elements (which are inorganic) and from essential amino
and fatty acids, which are required in larger amounts. The discovery of the vitamins began with
experiments performed by Hopkins at the beginning of the twentieth century; he fed rats on a
defined diet providing the then known nutrients: fats, proteins, carbohydrates, and mineral salts.
The animals failed to grow, but the addition of a small amount of milk to the diet both permitted
the animals to maintain normal growth and restored growth to the animals that had previously
been fed the defined diet. He suggested that milk contained one or more “accessory growth
factors” essential nutrients present in small amounts, because the addition of only a small amount
of milk to the diet was sufficient to maintain normal growth and development. The first of the
accessory food factors to be isolated and identified was found to be chemically an amine;
therefore, in 1912, Funk coined the term vitamine, from the Latin vita for “life” and amine, for
the prominent chemical reactive group. Although subsequent accessory growth factorswere not
found tobeamines, the name has been retained–withthe loss of thefinal“-e” to avoid chemical
confusion. The decision as to whether the word should correctly be pronounced “vitamin” or
“veitamin” depends in large part on which system of Latin pronunciation one learned – the
Oxford English Dictionary permits both.(Bender, 2003) There is no simple, overall statement
which embraces each and every biological function of the vitamins. This is scarcely surprising
because vitamins have themselves remarkably little in common, save for their organic nature and
the chance recognition oftheir biological importance at a particular time in history. In general
terms, vitamins act as either coenzymes or prohormones but, like all simplifications, this omits
certain other vital functions which they subserve. Originally they were regarded as being only
available in the diet (we now know that some can be synthesised in the body); to be ingested in
small amounts (but many other essential organic factors in the diet which are present in small
amounts are not classified as vitamins) and to be necessary as coenzymes (one is now best
regarded as a prohormone). In view of this great variety of activities the only reasonable way to
detail their biological functions is unavoidably to take each vitamin serially and separately. For
convenience they will be taken within their two separate chemical categories, first the four oil
soluble members of the group and then the water soluble ones. The latter group embraces several
compounds which are often grouped together under the term vitamin B-complex (or vitamin B-
group), the other member of the water soluble series being vitamin C. The vitamin B-complex
vitamins have perhaps the greatest level of biological similarity, for they playa vital role in
enzyme reactions which are necessary for carbohydrate, fat and protein metabolism. (Ottaway,
1993, p. 1)
2. Difinition
Vitamins are a group of organic compounds that are, in very small amounts, essential for the
normal functioning of the human body. They have widely varying chemical and physiological
functions and are broadly distributed in natural food sources. Thirteen vitamins are recognized in
human nutrition and these may be conveniently classified into two groups according to their
solubility. The fat-soluble vitamins are represented by vitamins A, D, E, and K; also included are
the 50 or so carotenoids that possess varying degrees of vitamin A activity. The water-soluble
vitamins comprise vitamin C and the members of the vitamin B group, namely thiamin (vitamin
B1), riboflavin (vitamin B2), niacin, vitamin B6, pantothenic acid, folate, and vitamin B12. This
simple classification reflects to some extent the bioavailability of the vitamins, as the solubility
affects their mode of intestinal absorption and their uptake by tissues. The solubility properties
also relate to the distribution of vitamins in the various food groups, and have a direct bearing on
the analytical methods employed. For many of the vitamins, biological activity is attributed to a
number of structurally related compounds known as vitamers. The vitamers pertaining to a
particular vitamin display, in most cases, similar qualitative biological properties to one another,
but, because of subtle differences in their chemical structures, exhibit varying degrees of
potency. Provitamins are vitamin precursors, that is, naturally occurring substances which are not
vitamins themselves, but which can be converted into vitamins by normal body metabolism.
Most of the vitamins are absolutely essential in the human diet because the body tissues cannot
synthesize them. Two notable exceptions are vitamin D and niacin. Cutaneous synthesis of
vitamin D depends on adequate exposure of the skin to sunlight, and the synthesis of niacin
depends on a sufficient intake of its amino acid precursor, tryptophan, bound within protein.
Plants have the ability to synthesize vitamins, except for vitamin B12, and serve as primary
sources of these dietary essentials .(Ball., 2006, p. 3)
2.1 Fat soluble vitamins

2.1.3 Vitamin E
The tocopherols are light yellow oils at room temperature.They are insoluble in water but are
readily soluble in nonpolar solvents. Being monoethers of a hydroquinone with a phenolic
hydrogen (in the hydroxyl group at position C-6 in the chromanol nucleus) with the ability to
accommodate an unpaired electron within the resonance structure of the ring (undergoing
transition to a semistable chromanoxyl radical before being converted to tocopheryl quinone),
they are good quenchers of free radicals and thus serve as antioxidants. They are easily oxidized,
however, and can be destroyed by peroxides, ozone, and permanganate in a process catalyzed by
light and accelerated by polyunsaturated fatty acids and metal salts. They are very resistant to
acids and (only under anaerobic conditions) to bases. Tocopheryl esters, by virtue of the blocking
of the C- 6 hydroxyl group, are very stable in air and are, therefore, the forms of choice as
food/feed supplements. Because tocopherol is liberated by the saponification of its esters,
extraction and isolation of vitamin E call for the use of protective antioxidants (e.g., propyl
gallate, ascorbic acid), metal chelators, inert gas environments, and subdued light. The UV
absorption spectra of tocopherols and their acetates in ethanol have maxima of 280–γ00 nm (α-
tocopherol, 292 nm); however, their extinction coefficients are not great. (Gerald F. Combs,
2008)
Natural (βR,4’R,8’R) α-tocopherol (RRR-α-T)

Chroman ‘head’ phytyl ‘Tail

2.1.2 Vitamin D

Vitamin D activity in foods is associated with several lipid soluble sterol analogs including
cholecalciferol (vitamin D3) from animal sources and ergocalciferol (vitamin D2) produced
synthetically (Figure 2.1). Both of these compounds are used in synthetic form for food
fortification. Cholecalciferol forms in human skin upon exposure to sunlight, and this is a
multistep process

involving photochemical modification of 7-dehydrocholesterol followed by nonenzymatic

isomerization.Because of this in vivo synthesis, the requirement for dietary vitamin D will
depend on the extent of exposure to sunlight. Ergocalciferol is an exclusively synthetic form of
vitamin D that is formed by commercial irradiation of phytosterol (a plant sterol) with ultra
violet (UV) light. Several hydroxylated etabolites of vitamin D2 and D3 form in vivo. The 1,25-
dihydroxy derivative of cholecalciferol is the main physiologically active form, and it is involved
in the regulation of calcium absorption and metabolism. 25-Hydroxycholecalciferol, in addition
to cholecalciferol, comprises a significant amount of the naturally occurring vitamin D activity in
meat and milk products. Fortification of most fluid milk products with either ergocalciferol or
cholecalciferol makes a significant contribution to dietary needs. Vitamin D is susceptible to
degradation by light, and this may occur in glass-packaged milk during retail storage. For
example, ∼50% of cholecalciferol added to skim milk is lost during 1β days of continual
exposure to fluorescent light at 4◦C. It is not known whether this degradation involves direct
photochemical degradation, a mechanism involving a photosensitizer yielding an active oxygen
species (e.g., 1O2), or as an indirect effect of light-induced lipid oxidation. Like other
unsaturated fat-soluble components of foods, vitamin D compounds are susceptible to oxidative
degradation. Overall, however, the stability of vitamin D in foods, especially under anaerobic
conditions, is not a major concern. (Srinivasan Damodaran, Kirk L. Parkin, Owen R. Fennema,
2008, pp.

2.1.3 Vitamin A
obtained from two sources: preformed retinol, usually in the form of retinyl esters hich can be
found in and therefore absorbed only from animal tissue and as carotenoids, mainly from plant
tissues. These carotenoids can be cleaved in the body to yield retinol. The main source of
carotenoid derived retinol is ȕ-carotene, though other carotenoids can also be converted to
retinol. Retinol is essential for growth and for the normal differentiation and development of
tissues. In many peripheral tissues retinol has a general metabolic effect, particularly in epithelial
cells where it is irreversibly converted into retinoic acid. The biochemical effect ofretinol (or
more correctly retinoic acid) in the peripheral tissues by means of which the integrity of the
epithelial cells is preserved is far from clear. It appears to be related to distortions in nitrogen
metabolism and to amino-acid balances within the tissues. This effect on nitrogen metabolism is
probably effected through RNA synthesis, perhaps by retinol serving as a carrier of sugars in the
formation ofspecific glycoproteins. This effect is seen particularly in various mucussecreting
tissues which become keratinised. Human vitamin Adeficiency, particularly in infants and
children can give rise to increased mortality during intercurrent infections. One of the most
obvious of the peripheral epithelial effects of a deficiency gives rise to the dryness of the
conjunctiva and the cornea (xerophthalmia) and subsequent ulceration which can lead on to
permanent eye damage. (Ottaway, 1993, pp. 1-2)
 Vitamin A (Retanol)

2.1.4 Vitamin K
Vitamin K occurs naturally in two main forms, vitamin K1 (phylloquinone), found in plants, and
vitamin K2 (menaquinone) present in bacteria and animals. A third compound, menadione, has
some vitamin K activity and is sometimes referred to as vitamin K3. Vitamins K1 and K2
contain phytyl and polyisoprenoid side chains, respectively (Figure 4). (J.M. Chesworth, T.
Stuchbury, J.R. Scaife, 1998) Phylloquinone (vitamin K1) is a product of plant origin, while
menaquinones (vitamin K2) of varying chain length are products of bacterial synthesis, mainly
by intestinal microflora. Phylloquinones occur in relatively large quantities in leafy vegetables
including spinach, kale, cauliflower, and cabbage, and they are present, but less abundant, in
tomatoes and certain vegetable oils. (Srinivasan Damodaran, Kirk L. Parkin, Owen R. Fennema,
2008, p. 466)
2.2 water soluble vitamin
The vitamins commonly considered in this group are thiamin, riboflavin, niacin, vitamin B6,
vitamin B12, folates, pantothenic acid, biotin and vitamin C. Again, many of these vitamins
occur naturally in more than one biologically active f~rm, and since these vitamins are water
soluble, they are generally more labile than the fat soluble vitamins. Most vitamins in this group,
with the exception of vitamin B12, are widely distributed in both animal and vegetable foods,
although the amounts present are often very small. Again, the development of more sensitive and
specific HPLC analysis techniques has improved the determination of many of the vitamins in
this group. Values from more conventional analytical methods, which generally appear in food
tables, are often in good agreement with values determined by HPLC, although HPLC provides a
more efficient analysis in most cases. (Ottaway, 1993, p. 28)
 2.2.1 Thiamin
Thiamin is present in practically all plant and animal tissues, however the content in many foods
is small, and food preparation may result in considerable losses. In most animal products,
thiamin occurs in a phosphorylated form (thiamin mono-, di- and triphosphates), with 80--85%
as the diphosphate, the active coenzyme form. In plant products, thiamin occurs predominantly
in the non-phosphorylated form. Assay methods require a suitable extraction procedure to release
the thiamin from the coenzyme in animal products. Foods rich in thiamin include dried brewer's
or baker's yeast (the former being the highest known source), pork, particularly pig's liver, cereal
germs, whole grains and wholegrain products, nuts and dried legumes. Thiamin is not found in
oils and fats, and is present at only low levels in green vegetables, fruit and seafood. In cereal
grains the thiamin is unevenly distributed, being low in the endosperm and high in the germ and
scutellum (the thin layer between the germ and endosperm), thus leading to substantial losses
from grain products on milling. (Ottaway, 1993, p. 28)

2.2.2 Riboflavin
 Riboflavin exerts its biological function through two flavin coenzymes,
inappropriately named flavin adenine dinucleotide (FAD) and flavin
mononucleotide (FMN). The coenzymes participate in oxidation– reduction
reactions in numerous metabolic pathways exemplified by the oxidation of reduced
nicotinamide adenine dinucleotide (NADH), the oxidation of succinate to fumarate
in the tricarboxylic acid cycle, and the oxidation of saturated fatty acyl coenzyme
A to a,ȕ-unsaturated fatty acyl coenzyme A in the b-oxidation of fatty acids. In all
of these reactions, the reduced flavin (FADH2 or FMNH2) is reoxidized by the
cytochrome electron-transport chain. In addition, flavin coenzymes are necessary
for specific reactions in the interconversion of vitamin B6 vitamers and folate
vitamers, and in the conversion of kynurenine to 3- hydroxykynurenine in the
synthesis of nicotinamide adenine dinucleotide (NAD) from tryptophan. (Ball.,
2006, p. 165)

2.2.3 Ascorbic acid

Ascorbic acid (L-AA) (Figure 7) is a carbohydrate-like compound whose acidic and reducing
properties are contributed by the 2,3-enediol moiety. This compound is highly polar; thus, it is
readily soluble in aqueous solution and insoluble in less nonpolar solvents.AAis acidic in
character as a result of ionization of the C-3 hydroxyl group (pKa1 = 4.04 at β5◦C). A second
ionization, dissociation of the C-2 hydroxyl, is much less favorable (pKa2 = 11.4). AA contains
two optically active centers at the C4 and C5 positions. l-Isoascorbic acid, the C-5 optical
isomer, and D-AA, the C-4 optical isomer (Figure 7), behave in a chemically similar manner to
AA, but these compounds have essentially no vitamin C activity. l-Isoascorbic acid (also known
as erythorbic acid) and AA are widely used as food ingredients for their reducing and
antioxidative activity (e.g., in the curing of meats and for inhibiting enzymatic browning in fruits
and vegetables), but isoascorbic ac id (and D-AA) has no nutritional value. (Srinivasan
Damodaran, Kirk L. Parkin, Owen R. Fennema, 2008, p. 467)

3. Food source of vitamins

All the vitamins required by man are available from the food supply, and to ensure an adequate
intake ofvitamins from food alone, it is advantageous to eat a variety of foods from both animal
and vegetable sources. Some vitamins are concentrated in a small number of foods, others are
widely distributed in nature, but may only occur in small quantities, and most vitamins are
considered to be micronutrients. The presence of vitamins in foodstuffs is variable depending on
a diverse range of factors, for example, the variety of plants, the season of the year, the growin
conditions, the breed, maturity and feeding oflivestock or the storage or treatment of the food. In
addition, the values commonly presented for the vitamin contents of foodstuffs in food
composition tables will vary depending on the analytical method used to assess the vitamin
content, as well as the sample of foods chosen for the analysis. It is important to appreciate that
samples of the same or similar foods may vary in composition, and that food table values
generally provide an average for use when analysing the diets of groups of people, rather than
individuals. One of the major differences between samples of similar foods will be variations in
the moisture content, which will strongly influence the proportion of all nutrients present. Where
vitamins are associated with aparticular macronutrient in foods, such as fat or protein, variations
in the content ofthese fractions will also affect the vitamin content. (Ottaway, 1993, p. 19) The
best approach to ensure you get a variety of vitamins, and in the proper amounts, is to adopt a
broad healthy diet. This involves an emphasis on fruits and vegetables, whole grains, beans and
legumes, low-fat protein, and dairy products. The good news is that many common foods contain
multiple mineral and vitamin sources, so it is easy to meet your daily needs from everyday
meals.
Here are some of the best foods for vitamins a the Harvard Medical School Special Heath
Report, Making Sense of Vitamins and Minerals: Choosing the foods and nutrients you need to
stay healthy

Vitamin Sources: 
Water soluble:
B-1: ham, soymilk, watermelon, acorn squash
B-2: milk, yogurt, cheese, whole and enriched grains and cereals.
B-3: meat, poultry, fish, fortified and whole grains, mushrooms, potatoes
B-5: chicken, whole grains, broccoli, avocados, mushrooms
B-6: meat, fish, poultry, legumes, tofu and other soy products, bananas
B-7: Whole grains, eggs, soybeans, fish
B-9: Fortified grains and cereals, asparagus, spinach, broccoli, legumes (black-
eyed peas and chickpeas), orange juice
B-12: Meat, poultry, fish, milk, cheese, fortified soymilk and cereals
Vitamin C: Citrus fruit, potatoes, broccoli, bell peppers, spinach, strawberries,
tomatoes, Brussels sprouts
 Fat soluble:
Vitamin A: beef, liver, eggs, shrimp, fish, fortified milk, sweet potatoes, carrots,
pumpkins, spinach, mangoes
Vitamin D: Fortified milk and cereals, fatty fish
Vitamin E: vegetables oils, leafy green vegetables, whole grains, nuts
Vitamin K: Cabbage, eggs, milk, spinach, broccoli, kale (Solan, 2016)

5. Methods used for analysis of vitamins:

5.1. High-performance liquid chromatography and Ultra performance liquid
chromatography - tandem mass spectrometer (UPLC-MS/MS)
 High-performance liquid chromatography
Many separation systems have been developed for the determination of folate vitamers using
HPLC including reversed-phase, ion pair, and anion exchange types. The developments in the
liquid chromatographic analysis of folates have been comprehensively reviewed by several
workers. The water-soluble nature of the folates, together with differences in ionic properties and
hydrophobicity, make these compounds well suited for either ion exchange, or reversed-phase
HPLC. Analysis of various naturally occurring folate polyglutamate derivatives can be
accomplished on hydrolysed extracts (as the corresponding folate monoglutamates), or as intact
poly-Ȗ-glutamyl folates. Monoglutamyl folates are usually separated by reversed-phase HPLC,
or by ion pair techniques. In reversed-phase separations, suppression or enhancement of the
ionization of functional groups by pH can effectively be used to regulate retention on the
column. The pH, ionic strength, and polarity of solvents are used to optimize the separation.
Usually low pH with or without gradient of the organic phase is used with C18 or phenyl
supports. (Andrѐ P. Leenheer, Willy E. Lambert, Jan F. Van Bocxlaer, 2000)
 Ultra performance liquid chromatography - tandem mass spectrometer (UPLCMS/MS) The
determination of vitamins in food represents a complex analytical problem. Vitamins naturally
present in foods at very low levels and in very complex matrixes. Furthermore, vitamins are
easily destroyed by strong acids or alkali; affected by many factors such as pH, heat, light, air,
extraction solvent, and time; and also affected by other food components. Despite the
development of several HPLC and LC–MS methods for vitamin analysis in food, there is still a
need for better resolution and sensitivity when analyzing complex food matrices containing
different vitamin derivatives. (Wilfried, 2006) Owing to its high resolution and sensitivity, as
well as its high sample throughput, UPLC is particularly attractive in the area of food analysis
where sample matrices are very complex. However, research in this field is still limited, and
much efforts and time are needed in order to provide more information and to improve the
existing methods. The content of vitamin C in food commodities (the sum of the contents of l-
ascorbic acid and dehydroascorbic acid) is used as an index of health-related quality of products,
since, as compared with other beneficial compounds, it is more sensitive for degradation by
processing and storage. Therefore, interest in the simultaneous analysis of these molecules has
increased greatly in food analysis. Spinola et al. determined the total vitamin C content in several
fruits and vegetables (lemons, passion fruits, papayas, strawberries, broccoli, green and red
peppers) using UPLC-photodiode array (PDA) system. (Novakova, L., Solich, P., 2008)

5.2. Gas chromatography

Gas chromatographic methods for assay of vitamin E were developed before the advent of LC.
GC methods applicable to determination of vitamin E in biologicals have been thoroughly
discussed by Nelis and colleagues100,102 and Lang and colleagues.101 Early methods were
hampered by the inability of packed-column chromatography to resolve ȕ- and Ȗ-tocopherols
and ȕ- and Ȗ-tocotrienols. In addition, packed-column chromatography was labor-intensive and
affected by interferences to both the tocopherols and the internal standard peaks, requiring
saponification to reduce the interferences and the use of correction factors to correct for
unremoved interferences.125 Lang and colleagues101 stated that through 1988, no packed-
column GC method had been developed to adequately resolve ȕ- and Ȗ-T or ȕ– and Ȗ-
tocotrienols (Ȗ-T3). Also, α-T3 and α-T were not well resolved. Development of capillary GC
solved many of the problems associated with packed-column chromatography. GC resolution of
ȕ- and Ȗ-T, and ȕ- and Ȗ-T3 can only be efficently accomplished as TMS derivatives on
capillary columns. Although LC is now universally accepted as an easy and accurate approach
for quantitation of vitamin E homologs, GC methods provide an alternative approach for analysis
of complex matrices. Such procedures usually rely on the linking of GC and mass spectrometry
(GC-MS).130–140 As an example, in 1998, Frega et al.130 identified and confirmed the 142
Vitamin analysis for the health and food sciences presence of several components of annatto,
including α- and ȕ-T3. Analysis of the total lipid fraction included saponification, treatment of
the extract with diazomethane to methylate free fatty acids, and silanization. Gas
chromatography was on the 30-m capillary column containing SPB-5 as the stationary phase.
Mottier et al.140 published a GC-MS and a LC-MS/MS method for quantitation of α-T and α-
tocopherolquinone in human plasma. For GC-MS assay, the analytes were converted to TMS-
derivatives and monitored in the selective ion mode. With the LC-MS/MS procedure, the
analytes were detected using multiple reactions monitoring after positive ESI. Both methods
used isotopically labeled (deuterated) forms of the analytes as internal standards. The GCMS and
LC-MS/MS-ESI procedures showed similar accuracy and within-day precisions. The
LC-MS/MS method was faster and more sensitive. (Ronald R. Etenmiller, Lin Ye, W. O. Landen
Jr., 2008)

5.3. liquid chromatograph-mass spectrometry

The feasibility of using reversed-phase liquid chromatography/diode array/tandem mass
spectrometry (LC–DAD–MS/MS) for a rapid and comprehensive profiling of fat soluble
vitamins and pigments in some foods of plant origin (maize flour, green and golden kiwi) was
evaluated. The instrumental approach was planned for obtainingtwomain outcomes within
thesamechromatographic run: (i) the quantitative analysis of ten target analytes, whose standards
are commercially available; (ii) the screening of pigments occurring in the selected matrices. The
quantitative analysis was performed simultaneously for four carotenoids (lutein, zeaxanthin, ȕ-
cryptoxanthin, and ȕ -carotene) and six compounds with fat-soluble activity (α-tocopherol, δ-
tocopherol, Ȗ -tocopherol, ergocalciferol, phylloquinone and menaquinone-4), separated on a
C30 reversed-phase column and detected by atmospheric pressure chemical ionization APCI
tandem mass spectrometry, operating in Selected Reaction Monitoring SRM mode. Extraction
procedure was based on matrix solid-phase dispersion with recoveries of all compounds under
study exceeding 78 and 60% from maize flour and kiwi, respectively. The method intraday
precision ranged between 3 and 7%, while the inter-day one was below 12%. The mild isolation
conditions precluded artefacts creation, such as cis-isomerization phenomena for carotenoids.
During the quantitative LC-SRM determination of the ten target analytes, the identification
power of the diode array detector joined to that of the triple quadrupole (QqQ) allowed the
tentatively identification of several pigments (chlorophylls and carotenoids), without the aid of
standards, on the basis of: (i) the UV–vis spectra recorded in the range of 200–700 nm; (ii) the
expected retention time; (iii) the two SRM ransitions, chosen for the target carotenoids but also
common to many of isomeric carotenoids occurring in the selected foods. (Alessandra Gentilli,
Fulvia Caretti, 2010)

6. Importance of Analysis
Vitamin analysis in food has a variety of purposes; it is used to provide quality assurance for
supplemented products; to study changes in vitamin content attributable to food processing,
packing, and storage; to provide data for food composition tables; and to check compliance with
contract specifications and nutrient labeling regulation. This section gives a short overview of
techniques for the analysis of the vitamin content in food and some of the problems associated
with these techniques. (Nollet, 200)
  CONCLUSION
After analyzing the vegetables and fruits it can be well
concluded that all of them contain one or the other
compounds vital for body functioning. It is observed that
carbohydrate is a predominant constituent while fats are
not present in most of the tested items. It is a natural
merit, as living organism require carbohydrate the most
common for generating energy. Among minerals,
presence of calcium is considerable as its present in all the
selected food items. Iron, magnesium and phosphorous
are also present sufficiently. Many other minerals form
constituents of vegetables and fruits, but in trace
quantities as body require them very less.
The results throw a light on significance of vegetables and
fruits as their constituents are vital compounds and
nutrients. The deficiency of these nutrients can lead to
various metabolic disorders. So, besides cereals, milk and
its products and non-vegetarian food items, vegetable and
fruits must be included in a daily balanced diet of all.
More effort is required to make everyone realizes their
significance for a healthy, disease-free, long lifestyle.

REFERENCE

 www.academia.edu
 NCERT Textbook
 Wikipedia
 www.seminarsonly.com