BIOCHEMICAL TESTS

SERUM ALBUMIN- INDEX OF PEM

AGE

SERUM
ALBUMIN
LEVELS
DEFICIENT
(HIGH RISK)

0-11

LOW (MEDIUM
RISK)

ACCEPTABLE
(LOW RISK)

<2.5

>2.5

1-5YRS

<2.8

<3.0

>3.0

6-17YRS

2.8

2.8-3.4

>3.5

ADULTS

<2.8

2.8-3.4

>3.5

PREGNANT (Ist
TRIMESTER)

<3.0

3.0-3.9

>4.0

PREGNANT (2nd
AND 3rd
TRIMESTER)

<3.0

3.0-3.4

>3.5

ESSENTIAL FATTY ACIDS

FATS- CONCENTERATED SOURCE OF ENERGY

SOURCE OF EFA
LINOLEIC ACID (18:2)
LINOLENIC ACID (18:3)
ARACHIODONIC ACID (20:4)
FUNCTIONS
STRUCTURAL/

FUNCTIONAL ROLES
PRECURSORS OF EICOSANOIDS; PROSTAGLANDINS
POTENT REGULATORS OF METABOLIC AND
PHYSIOLOGICAL FUNCTIONS

ESSENTIAL FATTY ACIDS

Assessment of Essential Fatty Acid Status
Alkali isomerization technique
Ratio

of Triene/ Tetraenes

Gas chromatographic technique

Measure

the Ratio of 20:3/ 20:4

Eichosatrienoic Acid (ETA)/ Arachiodonic Acid
(AA)
ETA non essential fatty acids- Oleic Acid
AA- EFA-Linoleic acid- desaturation and chain
elongation

ESSENTIAL FATTY ACIDS

Linoleic acid deficiency Ratio increases

Ratio= 0.2
Normal linoleic acid status
Disadvantages
N-3

PUFA intake is high

Desaturase activity reduced
Does not indicate- linolenic acid status

VITAMIN A

Serum/ plasma Vitamin A levels

Serum levels rises in women taking Oral
Contraceptives- due to mobilization of liver
stores
SERUM VITAMIN A
LEVELS
DEFICIENT
(HIGH RISK)

LOW (MEDIUM RISK)

ACCEPTABLE (LOW
RISK)

<20 g/dl

20-30g/dl

30g/dl

VITAMIN A
RELATIVE DOSE RESPONSE METHOD
BETTER

INDICATION OF VITAMIN STORES

%INCREASE IN SERUM VITAMIN A LEVELSMEASURED
AFTER 5 HOURSOF ORAL DOSE-450-1000g of
RETINOL
Post dose increase in serum vitamin A is inversely
proportional to Vitamin A status

VITAMIN A

Vitamin A deficiency- RBP accumulates in

liver
Availability of exogenous Vitamin A- release
oh Holo RBP
Raised circulating levels of Vitamin AMaintained depending upon the amount of
accumulated unbound RBP
RDR greater than 20%- suggestive of Vitamin
A deficiency

VITAMIN A
Modified version
Dehydro retinol (Vitamin A2) is administered
instead of Vitamin A
100g/Kg body weight children
2.5mg Adults
Only one post dose blood sample is required
Levels of vitamin A and A2 are measured
Ratio of A2/A>0.06= suggestive of Vitamin A
deficiency

VITAMIN D
INDICATOR OF VITAMIN D- 25-HYDROXY
CHOLECALCIFEROL (25HCC)
GUIDELINES

Acceptable

= >10ng/ml
Medium risk= 5-10ng/ml
Deficient, high risk= <5ng/ml

VITAMIN E

SERUM LEVELS OF VITAMIN E

RATIO OF VITAMIN- E TO TOTAL LIPIDS
SERUM VITAMIN -E/TOTAL LIPID = > 0.8
INICATIVE OF ADEQUATE STATUS
RED CELL FRAGILITY
MALONALDEHYDE FORMATION IN PRESENCE
OF HYDROGEN PEROXIDE- ARE USED AS
FUNCTIONAL INDICES OF VITAMIN E

VITAMIN K

Vitamin K required for carboxylation of Glutamic acid

residue for protein precursors of coagulation factors
and prothrombin
In Vitamin K deficiency- Precursors accumulate
PIVKAS-Proteins induced by Vitamin K deficiency
PIVKAS are absent- normal plasma
Measured
Spectrophotometry
Immunochemical
ELISA
Radioimmunoassay

methods

Vitamin K deficiency- prothrombin time -reduced

THIAMIN

Urinary thiamin
Thiochrome

method

HPLC
Microbiological

Assay
24 hour collection for 2 days
Early morning voided samples
Influenced by dietary intake
100g/24hoursor 65g/g creatinine

Blood (serum, RBC, WBC)levels of thiamin or

TPP

THIAMIN
Erythrocyte Transketolase (ETK) activity
coefficient
Xylulose -5 Phosphate+ Ribose-5-Phosphate

Sedoheptulose-7-Phosphate+
Glyceraldehyde -3-Phosphate
Xylulose -5 Phosphate +Erythrose-4-Phosphate
Fructose-6 Phosphate +
Glyceraldehyde -3-Phosphate

THIAMIN
ETK-AC=
ETK stimulated activity (+TPP)
ETK Basal activity (-TPP)
In B1 Deficiency-Basal activity Decreases and
Activity Coefficient Increases

Guidelines
Acceptable= <1.15
Medium Risk= 1.15-1.25
High Risk=>1.25

RIBOFLAVIN
Urinary riboflavin
Blood riboflavin
Erythrocyte Glutathione Reductase
(EGR)Test
GSSG+NADPH+H+
2GSH+ NADP+
EGR-AC=
EGR stimulated activity (+TPP)
EGR Basal activity (-TPP)
In B2 Deficiency-Basal activity Decreases and
Activity Coefficient Increases

RIBOFLAVIN
Guidelines
Acceptable= <1.5
Medium Risk= 1.5-1.7
High Risk=>1.7
NIACIN
Ratio of niacin metabolites
N-METHYL-2-PYRIDONE = <1= DEFICIENT
N1-METHYLNOCOTINAMIDE
Normal Range- 1-4

PYRIDOXIN

PLASMA PYRIDOXAL PHOSPHATE (PLP)

Normal

Range= 5-26ng/ml

URINARY VITAMIN B6
<20g/g

creatinine
PYRIDOXIC ACID- 0.5mg/day

ACTIVITIES OF ERYTHROCYTE TRANSAMINASES

& THEIR STIMULATION WITH PLP
TRYPTOPHAN LOAD TEST

PYRIDOXIN
ERYTHROCYTE TRANSAMINASES
L-Alanine+ - Ketoglutarate
L- glutamate
L-Aspartate+ - Ketoglutarate
Oxaloacetate+ L- glutamate
AC= Stimulated activity (+PLP)
Basal Activity (-PLP)
Guidelines
AspAT-AC<1.7-2.0
EAlaT-AC<1.25

Pyruvate+

PYRIDOXIN

Kynurinase enzyme- requires PLP as Cofactor

Vitamin B6 def.- activity of enzyme is
affected
Xanthurenic acid & 3-OH KynurenineElevated in Excretions
Oral Load Test-2-5 g Tryptophan
Normal 32Um Xanthurenic acid excreted in 8
hr collection after 2 g tryptophan load test
Pregnancy/ oral contraceptives- abnormal
values even when B6 status -normal

PYRIDOXIN

Tryptophan load test

FOLIC ACID

BLOOD FOLATE

SERUM FOLATE

RBC FOLATE

ACCEPTABLE

MEDIUM

HIGH RISK

>6

3-5.9

<3

>160

140-159

<140

FOLIC ACID

HISTIDINE LOAD TEST

URINARY

EXCRETION OF FORIMINOGLUTAMIC ACID

(FIGLU)
FIGLU- METABOLITE OF HISTIDINE
THF
FIGLU
GLUTAMIC ACID
NORMAL ADULTS- 20MG FIGLU IN 8HRS AFTER
HISTIDINE LOAD 0.26G/KG BODY WEIGHT
FOLIC ACID DEFICIENCY- FIGLU EXCRETIONINCREASES 5-10 TIMES

VITAMIN B12

BLOOD LEVELS
NORMAL ADULTS-200-900pg/ml
Deficient

(WHO/ FAO)= <80pg/ml

CUT OFF VALUES= <100pg/ml

SCHILLING TEST
MALABSORPTION

OF VITAMIN B12- DUE TO LACK

OF INTRINSIC FACTOR
PERNICIOUS ANAEMIA
MEASURING VITAMIN B12 ABSORPTION
WITH/WITHOUT INTRINSIC FACTOR

VITAMIN B12
METHYL MELONIC ACID EXCRETION
VIT.B12 DEFICIENCY- URINARY EXCRETION OF
METHYL MELONIC ACID (MMA) INCREASES
METHYL MELONYL CO.A METHYL MELONYL CO.A ISOMERASE B12
SUCCINYL CO.A
ORAL LOAD VALINE (5-10 g)
VALINE- PRECURSOR OF MMA

PANTOTHENIC ACID
Urinary Pantothenic Acid Excretion
NORMAL ADULTS-2mg/g

creatinine

Blood levels- 1g/ml

BIOTIN
BLOOD/PLASMA LEVELS
URINARY EXCRETION OF ORGANIC ACIDS
ACTIVITY OF BIOTIN DEPENDENT ENZYMES
NORMAL RANGE- BLOOD BIOTIN 120240mg/dl

ASCORBIC ACID

PLASMA ASCORBIC ACID LEVEL

LEUCOCYTE ASCORBIC ACID LEVEL
ACCEPTABLE

MEDIUM RISK

HIGH RISK

PLASMA
ASCORBIC ACID
(mg/dl)

0.3

0.2-0.29

<0.2

LEUCOCYTE
ASCORBIC ACID
(mg/dl)

15

8-15

<8

MINERALS

Calcium
Iron- Hb=13g/dl-Men; 12g/dl- Women
Iodine
Zinc
Copper
Selinium