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Biochemistry Lab Review

This document provides a summary of various biochemistry lab tests and procedures. It lists 34 different lab tests used to detect substances like carbohydrates, lipids, proteins, amino acids, and ketone bodies. It also summarizes several chromatography techniques used to separate biomolecules like proteins, amino acids, lipids, and sugars. These include gel filtration chromatography, ion exchange chromatography, thin layer chromatography, and paper chromatography. Additionally, it discusses techniques like radioimmunoassay, electrophoresis, dialysis, and colorimetry.

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76 views5 pages

Biochemistry Lab Review

This document provides a summary of various biochemistry lab tests and procedures. It lists 34 different lab tests used to detect substances like carbohydrates, lipids, proteins, amino acids, and ketone bodies. It also summarizes several chromatography techniques used to separate biomolecules like proteins, amino acids, lipids, and sugars. These include gel filtration chromatography, ion exchange chromatography, thin layer chromatography, and paper chromatography. Additionally, it discusses techniques like radioimmunoassay, electrophoresis, dialysis, and colorimetry.

Uploaded by

Femi Beauty
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOC, PDF, TXT or read online on Scribd
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BIOCHEMISTRY LAB REVIEW

MOLISH'S TEST: Presence of carbohydrates .1


BENEDICT'S TEST: Reducing sugars (glucose & sucrose) .2
SELIWANOFF'S TEST: Ketoses (fructose) .3
IODINE TEST: Polysaccharides & bile pigments .4
BIAL'S TEST: Pentoses (Ribose) .5

LECITHIN TEST: Presence of phospholipids .6


LIEBERMAN-BUCCHARD TEST: Presence of cholesterol .7
BROMINE TEST: Unsaturated fatty acids .8
ACROLEIN TEST: Presence of glycerol .9

BIUERT TEST: Peptide linkages .10


NINHYDRIN TEST: Amino acids .11

NDPH TEST: Ascrobic acid-2.4 .12


UV LIGHT TEST: Riboflavin .13

ALKALINE NA HYPOBRMITE: Urea .14


PHOSPHOTUNGSTIC TEST: Uric Acid .15
GAFF'S TEST: Creatinine .16

Proteins:17. SULPHOSALICYLIC ACID TEST


BRAD SHAW'S TEST: Benes Jones Proteins .18
GERHARD'S TEST: Acetoacedic acid (KETONE BODIES) .19
ROTHERA'S TEST: Acetoacetic and Acetone (KETONE BODIES) .20
FOUCHET'S TEST: Bile pigments .21
NITRIC ACID TEST: Bile pigments .22
EHRLICH TEST: Urobilinogen .23
HAV'S TEST: Bile salts .24
PETERKOFER'S TEST: Bile salts .25
HYPERCHOLESTEROLEMIA: Atheroscherosis .26
HYPOCALCEMIA: Tetany .27
HYPERURICEMIA: Allopurinol .28
GALACTOSEMIA: Mental Retardation .29
CREATINE PHOSPHOKINASE (CPK): Myocardial Infarction .30
CLINISTICK: Glucose .31
MOUSE-LIKE ODOR: Phenylketonuria .32
SULPHURONS: Cysinuria and Homocysinuria .33
SWEATY FEET: Isovaleric Acidaemia .34
CHROMATOGRAPHY is a separation technique that is used to
separate proteins & amino acids involving a mobile phase and stationary
phase and is exemplified by gel filteration

In GEL FILTERATION, biomolecules are separated according to


.difference in molecular weight

ION EXCHANGE CHROMATOGRAPHY is used for sequencing of


proteins

THIN LAYER CHROMATOGRAPHY is better than PAPER


:CHROMATOGRAPHY
Substances (ie. Lipids) that can't be separated by )1
paper are separated by chemicals
Faster )2
Sharper readings )3

PAPER CHROMATOGRAPHY used for identification of amino acids


:& sugars: 2 techniques

a) ASCENDING TECHNIQUE: the solvent moves vertically up


the paper by capillary action

b) DESCENDING TECHNIQUE: the solvent moves vertically


down by the help of gravity

c) MOBILE PHASE: solvent or gas used to separate component

d) STATIONARY PHASE: usually solid or liquid absorbent

e) SOLVENT FRONT: distance moved by mobile phase

f) RF VALUE: ratio of distance solute has traveled from point of


origin to distance traveled by solvent front

RADIOIMMUNE ASSAY requires antibodies and radioactive elements


for the quantitative measurements of hormones using a pure antigen of
known concentration

ELECTROPHORESIS can be used for purification of proteins &


nucleic acids involving the use of a polyacrylamide gel and provided an
electrical source

WESTERN BLOTTING for PROTEINS


SOUTHERN BLOTTING for DNA
NORTHERN BLOTTING for RNA

To separate a mixture of 20 amino acids the best technique is a TWO-


DIMENSIONAL ELECTROPHORESIS

:IN ELECTROPHORESIS the rate of migration depends on


NET CHARGE )1
SIZE & SHAPE )2
IONIC STRENGTH OF BUFFER )3
ELECTRICAL POTENTIAL )4

DIALYSIS involves diffusion across a POROUS MEMBRANE

COLORIMETRY is an instrument used for measuring the color


intensity of a substance (nm)

In gel filteration, biomolecule .1

DETERMINATION OF HEMOGLOBIN CONCENTRATION


.Unknown conc. = Unknown ABS. x Standard Conc
.Standard ABS

FACTORS AFFECTING ENZYMATIC REACTIONS


SUBSTRATE CONCENTRATION .1
TEMPERATURE .2
pH .3

NORMAL COMPOSITION OF URINE

INORGANIC CONSTITUENTS OF URINE


Chloride .1
Calcium .2
Phosphate .3
Ammonia .4
Sulphate .5

ORGANIC CONSTITUENTS OF URINE


Urea .1
Uric Acid .2
Creatinine .3

ABNORMAL CONSTITUENTS OF URINE


PROTEINS .1
REDUCING SUGARS .2
KETONE BODIES .3
BILE PIGMENTS .4
BILE SALTS .5

SERUM UREA: 15-45 mg/dl


URINE CREATINE: 0.4-1.8 g/24 hr
SERUM CALCIUM: 10mg/ dl
PLASMA PHOSPHATE: 2.5-5mg/ dl

GLUCOSE-TOLERANCE TEST

Glucose-level: 70-110 mg/dl


INSULIN lowers blood glucose level

GTT: a) patient should be on a normal diet prior to 3 days


b) Fasting 12-24 hours before
c) No smoking

NORMAL CURVE: No glucose in urine


DIABETIC CURVE: Level above 180 for temporary time (Glucosuria)
FLAT CURVE: Level fails to rise (Malabsorption)
LAG STORAGE CURVE: Liver Disease
RENAL GLUCOSURIA: Normal Glucose level because of low renal
threshold

:Mention the clinical condition in cases of elevated


a) Serum cholesterol: Atherosclerosis
b) Plasma uric acid: Hyperuricemia or Gout
c) Serum acid phosphatase: Prostate Cancer
d) Acetoacetate in plasma: Ketonemia
e) Blood glucose: Hyperglycemia or Diabetes Mellitus
f) Plasma urea: Renal Failure
g) Serum phenyl pyruvate: Phenylketonuria

"BE ABLE TO READ AND LABEL AND AUTORADIOGRAM "

Genotyping in DNA ANALYSIS (ie. Sickle cell mutation)

Be able to determine genotypes


?List the first 2 reaction in the PPP )1
What is the of NADPH in RBC's )2
Why are RBC more sensitive to G6P dehydrogenase deficiency more )3
?than hepatocytes
?Why in this case do jaundice and anemia develop )4
What are the types of jaundice? Name the similarities & differences )5
?What is the principle of PCR )6
?Mention two causes of hypercholesterolemia )7
?What is cholesterol level compatible with good health )8
?Give two different treatments to lower cholesterol level )9

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