Vitamin K plays an essential role in blood clotting and bone health. It exists in two primary forms, K1 and K2, with K2 having greater potency and a wider range of functions. Key sources of vitamin K include green leafy vegetables, fermented foods like natto, and bacteria in the intestines. Adequate vitamin K is required for blood clotting by enabling the liver to synthesize clotting proteins, as well as supporting bone mineralization and vascular health.

1. Analysis of VITAMINS
By:G.Pavani

2. Vitamins are organic nutrients that are required
in small quantities for a variety of biochemical
functions and which generally cannot be
synthesized in the body and must be supplied by
the diet

3. Objectives
• Vitamins are micronutrients
– Very small amounts are needed by the body (>1 gm)
– Very small amounts are contained in foods.
• Vitamins are essential.
– The roles they play in the body are very important.
– Most vitamins are obtained from the foods we eat.
– Some are made by bacteria in the intestine
– One is made in the skin
• There is no perfect food that contains all the
vitamins in the right amount.
Characteristics of Vitamins

4. Objectives
• Vitamins are non-energy producing
– They do not contain kcalories.
– They are involved in extracting energy
from the macronutrients.
• Some vitamins in foods are
precursors.
• Vitamins are classified according to
how soluble they are in fat or water.
Characteristics of Vitamins

5. What Are Vitamins?
• Tasteless organic compounds
• Required in small amounts
• Functions:
 Regulate Metabolism
 Promote Growth and Reproduction
Deficiencies can result in serious illnesses.

6. VITAL+
AMINE
Nutrients
VITAMINE
µg-mg
/day

7. Vitamins Can Be Destroyed by:
Cooking or Storage
• Exposure to Oxygen
–Water-Soluble Vitamins
–Vitamins A, E, and K
• Exposure to Ultraviolet (UV) Light
• Riboflavin (B2)
• Beta-carotene (A) and vitamin C

9. Vitamins Are Organic
Vitamins:
• All contain C, H and O.
• Can also contain some N and S.
• Each have a unique Chemical structure.
• Are each a singular unit.
• Perform numerous essential functions,
some with more than one role in
metabolism.

11. Pro-Vitamins Converted to Active Vitamins
• Provitamins - substances in foods that are not
directly ustable by the body.
– Converted to an active form once absorbed
– Example: beta-carotene vitamin A
• Preformed Vitamins
• Vitamins in foods in their active form.
• Do not need to be converted.

12. Classifying Vitamins:
Vitamins are classified based on solubility.
– Nine (9) Water-Soluble:
• B vitamin complex
• Vitamin C
– Four (4) Fat-Soluble:
• Vitamin A
• Vitamin D
• Vitamin E
• Vitamin K

14. Water soluble vitamins Fat soluble vitamins
Solubility Water soluble Fat soluble
Absorption Simple Along with lipids
Storage *No storage Stored in liver
Excretion Excreted Not excreted
Excess intake Nontoxic Toxic
Deficiency manifest rapidly Manifest slowly
Treatment Regular dietary supply Single large dose
Difference b/w water soluble & fat soluble vitaminsVITAMINS

15. The many roles of Vitamins in maintaining Health

16. Fat soluble vitamins

17. Fat soluble

18. vitamin A: as retinol or esters of retinol in the form of
retinyl acetate or retinyl palmitate,

19. Vitamin A
• Vitamin A is a Family of fat-soluble Retinoids:
Retinol, Retinal, Retinoic Aacid
Retinol Retinal Retinoic Acid
• Animal foods
• Stored in Body
• Function:
Reproduction
• Found in plants
• Retinal = Vision
• Retinoic = Regulates growth

20. • Other names
– Preformed Vitamin A – retinyl esters
• Retinol, Retinal, Retinoic acid
• Sources: animal foods, fortified foods,
pharmaceutical supplements
– Provitamin A - Precursors=carotenoids
• Alpha and Beta-carotene and lycopene
• Sources: plant foods
Vitamin A

21. • Chief functions in the body
– Vision
– Maintenance of cornea, epithelial cells,
mucous membranes, skin
– Bone and tooth growth
– Reproduction
– Immunity
– Antioxidant effect of beta-carotene
Vitamin A

23. • Retinol
– Fortified milk, butter
cheese, cream
– Fortified margarine
– Eggs
– Liver
Vitamin A Sources

24. Carotenoids
Are pigmented molecules in plants that give
intense orange, yellow and even red colors to a
variety of fruits and vegetables.
The carotenes that act as antioxidants are:
1) alpha carotene
2) beta carotene
3) lycopene

25. Beta-Carotene
A very important Carotenoid in Human Nutrition; a
pigment deep orange/yellow (e.g. carrots and pumpkin).
The human body has 2 functions for carotenes:
1. It turns it into Vitamin A 40% of carotenes
2. It turns it into an antioxidant. 60% of carotenes

26. – Dark leafy green
vegetables, spinach,
broccoli
– Deep orange veggies
• Carrots, pumpkin,
squash, sweet potato
– Deep orange fruits
• Apricots, cantaloupe
Beta-carotene Sources

27. Lycopene
Bright Red carotene phytochemical.
Tomatoes are a perfect example.
Watermelon, papayas, guava
pink grapefruit.
Also in asparagus, brown beans
and parsley

28. • Night blindness
• Xerosis (corneal drying)
• Bitot’s spots
• Karatomalacia
• Xerophtalmia
• Hyperkaratosis
• Impaired immunity
Vitamin A Deficiency

29. • Keratinization
– Vitamin A
deficiency
symptom
Vitamin A Deficiency

30. • Acute toxicity symptoms
– Blurred vision
– Nausea, vomiting, vertigo
– Increase of pressure inside skull,
mimicking brain tumor
– Headaches
Acute Toxicity

31. • Chronic toxicity symptoms
– Increased activity of osteoclasts
causing reduced bone density
– Liver abnormalities
– Birth defects
Chronic Toxicity

32. Standard preparation— Dissolve an accurately weighed quantity of USP Vitamin A
RS in n-hexane, and dilute quantitatively, and stepwise if necessary, to obtain a
solution having a known concentration of about 15 µg of retinyl acetate per mL.
Assay of vitamin A ester (retinyl acetate or retinyl palmitate) by HPLC as per USP
Extraction : Weigh (10g or 10mL) and finely powder (if solid) the sample. Transfer an
accurately weighed portion of the powder to a container having a plastic container
with cap, add 10 mL of dimethyl sulfoxide and 15 mL of n-hexane, and shake for 45
minutes (water bath maintained at 60 deg).
Separation: Centrifuge at 3000 rpm for 10 minutes, and transfer the hexane layer by
means of a pipete to a 100-mL volumetric flask.

33. Add 15 mL of n-hexane to the dimethyl sulfoxide layer, shake thoroughly for 5 minutes,
and transfer the hexane layer by means of a pipette to the 100-mL volumetric flask.
Repeat this extraction with three additional 15-mL portions of n-hexane.
Dilute the extracts in the volumetric flask with n-hexane to volume, and mix.
Chromatographic conditions:
Mobile phase is n-hexane. The liquid chromatograph is equipped with a 325-nm
detector and a 4.6-mm × 15-cm column that contains 3-µm packing L8. The flow rate is
about 1 mL per minute.

34. in which 0.872 is the factor used to convert retinyl acetate, obtained from USP
Vitamin A RS, to its retinol equivalent; C is the concentration, in mg per mL, of
USP Vitamin A RS in the Standard preparation; D is the dilution factor, in mL,
for the Assay preparation; rS and rU are the peak areas of the all-trans retinyl
ester obtained from the standard and Assay preparation and the.
calculate the quantity, in mg, of vitamin A

35. vitamin D:cholecalciferol or ergocalciferol

36. Vitamin D
The “Sunshine Vitamin"
Need two things!
1) Ultraviolet (UV) rays.
2) Form of Cholesterol in the skin.
All your vitamin D needs can be met by exposure to sunlight : )
Considered a prohormone - Found in two forms:
1) Cholecalciferol (vitamin D3) = in our skin and in animal foods
2) Ergocalciferol (vitamin D2) = in plants & dietary supplements

37. • Other names
– Calciferol
– 1,25-dihyroxy vitamin D (calcitriol)
– Animal version: vitamin D3 or
cholecalciferol
– Plant version: vitamin D2 or ergocalciferol
– Precursor is the body’s own cholesterol
Vitamin D

38. The Chemical Structure of Vitamin D

40. • Chief functions in the body
– Mineralization of bones
• raises blood calcium and phosphorus by
increasing absorption from digestive tract
• withdrawing calcium from bones
• stimulating retention by kidneys)
Vitamin D

41. ASSAY FOR CHOLECALCIFEROL / ERGOCALCIFEROL as per USP (VITAMIN D)
Standard preparation— Dissolve an accurately weighed quantity of USP Cholecalciferol
RS or USP Ergocalciferol RS in n-hexane, and dilute quantitatively, and stepwise if
necessary, to obtain a solution having a known concentration of about 2 µg per mL.
Extraction : Weigh (10g or 10mL) and finely powder (if solid) the sample. Transfer an
accurately weighed portion of the powder to a container having a plastic container
with cap, add 10 mL of dimethyl sulfoxide and 15 mL of n-hexane, and shake for 45
minutes (water bath maintained at 60 deg).
Centrifuge at 3000 rpm for 10 minutes, and transfer the hexane layer by means of a
pipet to a 100-mL volumetric flask.

42. Add 15 mL of n-hexane to the dimethyl sulfoxide layer, shake thoroughly for 5
minutes, and transfer the hexane layer by means of a pipet to the 100-mL volumetric
flask. Repeat this extraction with three additional 15-mL portions of n-hexane
Dilute the extracts in the volumetric flask with n-hexane to volume, and mix.
The liquid chromatograph is equipped with a 265-nm detector and a 4.6-mm × 15-cm
column that contains 3-µm packing L8. The flow rate is about 1 mL per minute.
C is the concentration, in µg per mL USP Cholecalciferol RS or USP Ergocalciferol RS in
the Standard preparation
1.09 is a correction factor
D is the dilution factor, in mL, for the Assay preparation
Mobile phase— Prepare a filtered and degassed mixture of n-hexane and isopropyl
alcohol (99:1).

43. Vitamin-E
Alpha tocopherol or alpha tocopheryl acetate or alpha
tocopheryl acid succinate , phytonadione

44. Vitamin E sources

45. functions

46. Vitamin E Toxicity and Deficiency
No risk of too much vitamin E from food sources!
Too much synthetic form may risk of a hemorrhage.
Deficiency Symptoms in Adults
Mild Anemia, Fragile RBCs
Neurological Damage
Disorders related to Reproduction and Infertility
Age spots, Cataracts
Muscle, liver, bone marrow brain abnormalities
Decrease in Sex Drive
Significantly more free radical damage to cell membranes

47. Assay for vitamin E (alpha tocopherol, alpha tocopheryl
acetate, or alpha tocopheryl acid succinate)
Standard preparation— Dissolve an accurately weighed quantity of USP Alpha
Tocopherol RS, USP Alpha Tocopheryl Acetate RS, or USP Alpha Tocopheryl Acid
Succinate RS in methanol, and dilute quantitatively with methanol to obtain a
solution having a known concentration of about 2 mg per mL.
Extraction : Weigh (10g or 10mL) and finely powder (if solid) the sample. Transfer an
accurately weighed portion of the powder to a container having a plastic container
with cap, add 10 mL of dimethyl sulfoxide and 15 mL of n-hexane, and shake for 45
minutes (water bath maintained at 60 deg).
Centrifuge at 3000 rpm for 10 minutes, and transfer the hexane layer by means of a
pipet to a 100-mL volumetric flask.

48. Add 15 mL of n-hexane to the dimethyl sulfoxide layer, shake thoroughly for 5
minutes, and transfer the hexane layer by means of a pipet to the 100-mL volumetric
flask. Repeat this extraction with three additional 15-mL portions of n-hexane
Dilute the extracts in the volumetric flask with n-hexane to volume, and mix & inject to
HPLC.
The liquid chromatograph is equipped with a 254-nm detector and an 8-mm × 10-cm
column that contains 5-µm packing L1. The flow rate is about 2 mL per minute.
Mobile phase— Dilute 10 mL of phosphoric acid with water to 1000 mL to obtain
Solution A. Prepare a filtered and degassed mixture of methanol and Solution A (95:5).
C is the concentration, in µg per mL USP Cholecalciferol RS or USP Ergocalciferol RS in
the Standard preparation
26.5 is a correction factor
D is the dilution factor, in mL, for the Assay preparation

49. Vitamin-K

50. • Other names
– Phylloquinone
– Manaquinone
– Menadione
– Naphthoquinone
• 2001 AI
– Men: 120 g/day
– Women: 90 g/day
Vitamin K

51. • Bacterial synthesis
in GI tract
• Leafy green
vegetables
• Cruciferous
vegetables
• Liver
• Milk
Vitamin K Sources

52. 1) Green Leafy Vegetables (Kale) ½ c: 444 mcg (over 100% DV)
2) Natto (Fermented soy) 2 oz: 500 mcg (over 100% DV)
3) Spring Onions (Scallions) ½ c: 103 mcg (over 100% DV)
4) Brussels Sprouts ½ c: 78 mcg (98% DV)
5) Cabbage ½ cup: 82 mcg (over 100% DV)
6) Broccoli ½ c: 46 mcg (58% DV)
7) Dairy (Fermented) ½ c: 10 mcg (10% DV)
8) Prunes ½ c: 52 mcg (65% DV)
9) Cucumbers 1 med 49 mcg (61% DV)
10) Dried basil 1 Tbsp: 36 mcg (45% DV)
Natural Foods Rich in Vitamin K1 and K2

53. • Bacteria in intestines produce ~75% of
Vitamin K absorbed by body daily
• Vitamin K not stored in body
• Vitamin K needs to be supplied daily
• Absorption dependent on healthy liver
and gall bladder
Vitamin K

54. • Family of vitamins
– Naturally found in primarily two forms
• K1 and K2
• K3 simpler form; synthetically created
• Identified by German scientists
• Required for normal blood clotting
– Named “K” for German word for “clot”
• No Tolerable Upper Limit
Vitamin K

55. Vitamin K types
• 2 forms of vitamin K
– Menaquinone from intestinal bacteria. K1
– Phylloquinone is found in green plants. K2

56. • K1 produced by plants we eat
• K2 produced by bacteria in intestine;
converted from K1
• K2 more potent (15x); more active; and
wider range of activities
– Better absorbed; longer biological activity
– Predominant form found in body tissues
– Used preferentially by all tissues but liver
Vitamin K

57. • Vitamin K1 produced by plants and algae
– Broccoli, kale, chard; plant oils like canola and
soybean
• Hydrogenated soybean oil has ineffective K
• Vitamin K2 produced by bacteria in gut
– Food Sources: fermented soybean (Natto);
dairy products, egg yolk
Vitamin K Sources

58. • Chief functions in the body
– Synthesis of blood-clotting proteins and bone
proteins that regulate blood calcium
• Synthesis of bone proteins that regulate
blood calcium, prevent bone loss
• Integration of calcium into bones
• Prevent calcium deposition in blood vessels
(vascular calcification)
• Maintain blood vessel elasticity
Vitamin K

59. Vitamin K: Other Functions

60. Functions of Vitamin-K

62. WATER Soluble vitamins analysis
vitamins analysis