14-vitamine Flashcards

1
Q

define vitamin

A

Vitamins are defined as relatively low-molecular weight compounds which are required in small quantities for normal metabolism.

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2
Q

Health benefits Regulatory requirement Addition of ____ to fluid milk is a legal requirement under Health Canada’s Food and Drugs Act

A

Vitamin A, Vitamin C and Vitamin D

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3
Q

how is vitamin calssified?

A

water soluble BC

fat soluble ADEK

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4
Q

why analysis vitamin?

A

 Nutritional quality of food because of the health benefits Humans do not synthesize vitamin and therefore need to them from food
obtain
 Regulatory requirement
Addition of Vitamin A, Vitamin C and Vitamin D to fluid milk
is a legal requirement under Health Canada’s Food and Drugs Act
 One of the determinants of shelf-life of food
Vitamins are relatively labile compounds that are affected by factors such as heat, light, air, pH, other food components
 Indicate effect of food processing/packaging/and storage on the nutritional quality of food

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5
Q

what is the vitamin unit?

A

Usually expressed in units of mg or μ g per tablet or food serving

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6
Q

how can vitamin expressed?

A

international units (IU), US Pharmacopeia (USP) units , and % Daily Value (DV).

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7
Q

what is IU? what does it based on? what does it used for?

A

The IU is a unit of measurement for the amount of a substance, based on measured biological activity or effect. It is used for vitamins, hormones, vaccines, and similar biologically active substances.

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8
Q

The precise definition of 1 IU ______ substance to substance.

A

differs from

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9
Q

3 ex of IU

A

1 IU of vitamin A is the biological equivalent of 0.3 μg retinol, 0.6 μg β -carotene, and 1.2 μg of other pro-vitamin A active carotenoids (e.g., α -carotene and β -cryptoxanthin).
 1 IU of vitamin C is the biological equivalent of 50 μg L -ascorbic acid.
 1 IU of vitamin D is the biological equivalent of 0.025 μg cholecalciferol/ergocalciferol.

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10
Q

the pathway of sample processing

A

sample extraction get extract

  1. clean up get purified extract
  2. quantitative and qualitative analysis
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11
Q

why analysis of vitamins is performed by different chemical, physical, and biological methods.

A

Vitamins belong to different classes of organic substances

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12
Q

vitamin assays in most situation involve the extraction of a vitamin from_____ to analysis.

A

biological matrix prior to analysis.

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13
Q

which treatment used to extract vitamin

A

heat, acid, alkali, solvent, enzyme

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14
Q

what is the function of extraction procedure?

is it specific/non specific for vitamin?

A

extraction procedures are specific for each vitamin and designed to stabilize the vitamin.

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15
Q

In some case, some procedures are applicable to the ______ of more than one vitamin.

A

combined extraction

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16
Q

Typical extraction procedures :

Ascorbic acid?

A

Cold extraction with metaphosphoric acid/acetic acid

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17
Q

Typical extraction procedures :B1,B2

A

Boiling or autoclaving in acid plus enzyme

treatment.

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18
Q

Typical extraction procedures :B3

A

Autoclaving in acid (non-cereal products) or alkali (cereal products).

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19
Q

Typical extraction procedures :B9

A

Enzyme extraction with α –amylase, protease and γ -glutamyl hydrolase.

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20
Q

Typical extraction procedures :AED

A

Organic solvent extraction, saponification, and re- extraction with organic solvents.

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21
Q

what is the chanllenge of determine VD in milk?

A

 Relatively low concentration
 Poor stability in response to heat, light, oxidation,
 Possible interference from milk components (e.g., fat, emulsifiers, amino acids,
proteins, sterols, and minerals)

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22
Q

what is the key in VD determination

A

sample preparation

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23
Q

procedure of extract VD

A

solvent extraction
saponification
re-extraction

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24
Q

3 type of vitamin assay

A

bioassay
microbiological assay
physicochemical assay

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25
Q

physicochemical contain?

A

spectrophotometric methods.

  1. 2- fluorometric methods.
  2. 3- chromatographic methods.
  3. 4- enzymatic methods.
  4. 5- immunological methods. 3.6- radiometric methods.
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26
Q

bioassay involve?

A

animal

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27
Q

micorbiological assay use?

A

protozoan, bacteria yeast, organism

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28
Q

Bioassays at the present are used only for the analysis of ?

A

vitamins B12 and D.

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29
Q

Vitamin D assay using__models

A

rat

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30
Q

Line test for Vit D is based upon?

A

the cure of experimentally induced rickets in rats during a specific period, as measured by the calcification of bones, the extent of calcification being determined by the “line test’’

31
Q

Bone sections (logitudinal) are stained with___

A

silver for observing calcification

32
Q

how to define amount of VD?

A

Compare the extent of this line formation with that of reference group receiving oil containing

33
Q

Microbiological assays are limited to the analysis of __. The methods are very sensitive and specific for __.

A

water-soluble vitamins.

each vitamin.

34
Q

The growth of microorganisms is ____to their requirement for a specific vitamin.

A

proportional

35
Q

The vitamin tested will act as the__?

A

growth limiting nutrient.

36
Q

how to test growth limiting nutrient?

A

The growth of a certain microorganism in an extract of a vitamin-containing sample is compared against the growth of this microorganism in the presence of known quantities of that vitamin.

37
Q

Bacteria, yeast, or protozoans are used as test ___?

A

organisms

38
Q

how can growth be measured?

A

turbidity , acid production, gravimetric, or

by respiration .

39
Q

With bacteria and yeast, _____ is the most commonly

employed system

A

turbidimetry

40
Q

procedure of analysis Niacin

A

1- Weigh the sample.
2- Add 1N H2SO4.
3-Autoclave 30min at 121-123 0C.
4-Filter the mixture, discard and residue.
5- To the filtrate add Difco Niacin Assay Medium (assay medium is devoid of Niacin).
6-Autoclave 10min at 121-123 0C.
7- Add Lactobacillus plantarum ATCC 8014. 8-Incubate at 37 0C for 16-24hr.
9- Measure the optical density and compare with that of bacteria grown in medium containing known amount of the vitamin (Niacin)

41
Q

procedure of analysis folate

A

1- Weigh the sample (2g).
2- Add 50ml buffer
3-Add α-amylase, protease, and conjugate and allow for the digestion of the matrix components
4-Deactivate enzymes by boiling the mixture for 5min.
5-Filter the mixture and discard the residue
6- Add the filtrate with bacterial growth medium
7-Autoclave for 10min at 121-123 0C.
8- Add Lactobacillus casei (spp. rhamnosus) ATCCTM 7469 .
9-Incubate at 37 0C for 20-24hr.
10-Measure the optical density and compare with that of bacteria grown in medium containing known amount of the vitamin (folate)

42
Q

VA is sensitive to?

A

UV air high temp, moisture

43
Q

what method is used to measure VA activity

A

only HPLC

44
Q

Principle of VA activity measurement

A

The test sample is saponified with ethanolic KOH, vitamin A (retinol) is extracted into organic solvent and then concentrated. Vitamin A isomers – all-trans- retinol and 13-cis-retinol – levels are determined by HPLC on a silica column.

45
Q

steps of VA

A

1- Transfer 40 ml of ready-to-use formula or fluid milk to a 100-ml digestion
flask containing a stirring bar.
2- Add 10 ml of ethanolic pyrogallol (absorb oxygen) solution and 40-ml ethanolic KOH
(for saponification of lipids—>Potassium salts of fatty acids and glycerol).
3-stir at room temperature for 18 hr.
4-Extract vitamin A with hexane:diethyl ether mixture
5- Analyze the organic layer using HPLC for Vit A.

46
Q

what is the principle of analysis VC?

A

L-ascorbic acid is oxidized to L-dehydro-ascorbic acid by the 2,6-dichloroindophenol (DCP).

47
Q

what is 2,6 DCP

A

oxidation– reduction indicator dye,

48
Q

Presence of ascorbic acid in solution/juice keep the pH ___

A

acidic

49
Q

DCP is___ colored dye in acidic solution.

A

red

50
Q

DCP specifically/nonspecifically oxidize ascorbic acid

finally turn ___ color?

A

specifically

colorless

51
Q

the end point of DCP is ___ color?

A

rose-pink

52
Q

VC is susceptible to?

A

oxidative deterioration that is accelerated by high pH and the presence of ferric and cupric ions.

53
Q

If significant amounts of ferric Fe, cupric Cu ions are present in the sample to be analyzed,___ need to add with the extraction

A

EDTA

54
Q

what is EDTA

A

chelating agent?

55
Q

titrant DCP will ____ from ___ to ___ color. as L-ascorbic acid which is ___ color will change to L-dehydroascrobic acid which is ___ color

A

reduce
red to colorless
colorless
brown

56
Q

Thiamine use which method?

A

Thiochrome fluorometric

57
Q

write down B1 procedure

A
  • Extraction of thiamine in 0.1N H2SO4with dilute acid
  • Enzymatic hydrolysis of thiamine’s phosphate-esters (using diesterase) at pH 4.0-4.5 in presence of 1.2 M sodium acetate
  • Chromatographic cleanup (i.e., purification),
  • Oxidation to thiochrome
  • Measurement using fluorescence intensity
58
Q

what does enzymatic do?

A

hydrolysis of thiamine’s phosphate-esters

59
Q

what does the pH need for thiochrome fluorometric?

A

4-4.5

60
Q

what solution need to hydrolysis thiamine’s phosphate-ester?

A

sodium acetate

61
Q

how to oxidation to thiochrome?

A

using alkaline potassium ferricyanide

62
Q

what need to be considered as selection of method?

A
  1. Method accuracy and precision (Consider the use of official methods)
  2. The need for bioavailability information
  3. Time and instrumentation requirements
  4. Personnel requirements
  5. The type of biological matrix to be analyzed
  6. The number of samples to be analyzed
  7. Regulatory requirements – Must official AOAC International methods be used?
  8. Suitability of the method (Applicability of microbiological assays is limited to water-soluble vitamins (most commonly niacin, B12, and pantothenic acidB5)).
63
Q

Cold extraction with metaphosphoric acid/acetic acid. extract?

A

ascorbic acid

64
Q

Autoclaving in acid (non-cereal products) or alkali (cereal products).extract?

A

B3 Niacin

65
Q

Boiling or autoclaving in acid plus enzyme

treatment. extract?

A

B1 or B2

66
Q

Organic solvent extraction, saponification, and re- extraction with organic solvents.extract?

A

A.E.D

67
Q

Enzyme extraction with α –amylase, protease and γ -glutamyl hydrolase. extract?

A

folate

68
Q

B12, D using ___ assay?

A

bioassay

69
Q

B3,B9 using___ assay?

A

microbiological

70
Q

VA using___ method?

A

physicochemical– chromatographic

only HPLC

71
Q

water soluble vitamin using __ assay?

A

microbiological

72
Q

B1 using___ method?

A

physicochemical– thiochrome fluorometric

73
Q

VC using ___ indicator?

A

DCP