Lab 10 - Examination of lipids Flashcards by Hanne Hansen

What are lipids?

Compounds which are INSOLUBLE in WATER but,

SOLUBLE in FAT solvents

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How do lipids differentiate?

In chemical structures

Into different groups - 2 main groups

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What are the two different main groups of lipids?

Simple Lipids
Derivated lipids (Lipoids)

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What is Simple lipids, where are they found and what is their main function?

Esters of FA with GLYCEROLL
Generally found in organism as TRIACYLGLYCEROLS in greater amount.
ENERGY STORAGE, transferd by the blood circulation to the fat tissue to be stored as DEPOT fat.
Or transported from the DEPOT to the user organs
(Free FA: FFA)
or NEFA

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Where can the FA be detected?

In the blood plasma

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What is characteristic of the FA in plants?

High proportion of unsaturated FA is characteristic

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Almong Lipid, there is a steroid that is important?

Cholestrol

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What is Cholestrol and where is it synthesized ?

The precursor of the STEROID hormones and BILE ACIDS
Synthesized in the LIVER
Also a constituent of the biological membrane

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What is phosphatiedes, and where does it occur?

Such as Lectin

Occur mainly as a constituents to

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Where is the long chain of FA transported?

Fat DEPOT –> User organs (muscle, organs) by the blood

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What do free FA bind to?

Albumin with secondary bonds

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What is the FA detected by?

Cu(II)Sulphate
CuSO4
Cu(II)Nitrate

From the albumin

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What will produced copper salt dissolve in?

Chloroform

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The nr of Cu2+ ions in chloroform = Nr of FA, but how can we measure it?

By Diethyldithiocarbamate by photometry

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How do you measure the quantitative estimation of the FA?

The nr of Cu2+ ions in chloroform = Nr of FA

By Diethyldithiocarbamate by photometry

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What are the important materials of the measurement of the quantitative estimation of the FA?

Concentrated Blood Plasma
Copper(II) Reagent
Na-diethyldicarbamate
Choloroform
Spectrometer
Tube with glass stopper

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What does the Copper(II) Reagent consist of?

270 ml Triethanol amine (2mmol/l)
30ml Acetic Acid (1mol/l)
300ml copper sulphate solution (0,63 mol/l)

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What can you observe in the measurement of the quantitative estimation of the FA?

3 phases:
Upper: foamy, blue, free Cu2+ ions –> not taken part)
Middle: Proteins (pipetted out)

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Clinical importance of the measurement of the quantitative estimation of the FA?

Elevated free FA conc occurs due to increased lipolysis in case of energy deficiency, such as….
Starvation and Ketosis

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Where can cholesterol be found?

Almost in every animal tissue = cause its a constituent of the cell membranes.

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What does Cholestrole Precurs?

Synthesis of different
STEROID HORMONES
BILE ACIDS
Imp-role in fat digestion

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How do we measure the cholesterol concentration ?

Measured from the blood plasma with spectrophotometry

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What is the principle of measuring the cholesterol conc . from the BP with spectrophotometry?

In the presence of GLACIAL ACETIC ACID and SULPHURIC ACID, FeCl3 will oxidize cholesterol to form new unsaturated double bonds.

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How many of the new cholesterol unsaturated double bonds are able to bind to each other?

Two go such oxidized cholesterol molecules bind to each other and produce CONJUGATED BONDS

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When two oxidized cholesterol molecules bind to each other, what does it produce?

CONJUGATED BONDS

Which color does the new complex of CONJUGATED form?

PURPLE COLOUR, color measured by spectrophotometer

When measuring the total cholesterol content in blood, what are the important materials?

- Blood plasma 3X diluted - FeCl3 solution: 10g FeCl3 solution to 200ml conc sulphuric acid - Glacial acetic acid

Does the total measurement of cholesterol have a minor or major clinical imp. in vet.med?

Minor

Which diseases can the total cholesterol measurement provide a useful help in diagnostics of what ?

Hyperlipidaemia Some Hepatic diseases (Hepatopathies) Hyperadrenocorticism Hyperthyreosis

What are the normal amount of cholesterol in dogs?

2,5-8 mmol/l

What are the normal amount of cholesterol in cats?

2-6,5 mmol/l

What are the normal amount of cholesterol in herbivores?

1,5-5 mmol/l

Determination of triglyceride in blood plasma is carried out of which enzymatic reactions?

Lipoprotein lipase Glycerol kinease Gly-3-p oxidase (GPO) Peroxidase (POD)

What are Triglycerides transported in?

CHYLOMICRONS, and different types of LIPOPROTEINS in the Bloodplasma

What are the different LIPOPROTEINS?

VLDL LDL IDL HDL

Triglycerides transported in lipoproteins are hydrolysed to.... by which enzyme

Glycerol and FA by | Lipoprotein lipase

Glycerol Kinease enzyme converts?

Glycerol ---> Glycerol-3-P by using up one ATP

Glycerol-3-phosphate oxidase (GPO) oxidizes....

Glycerol-3-P to Dihydroxyacetone-P

GPO belonges to which group?

Anaerobe dehydrogenase, meaning that the produced FADH2 passes the HYDROGENS directly to the OXYGEN of the air, and H2O2 is RELEASED.

Triglycerides + 3H2O ----->

Glycerol + 3 FA | Enzyme: Lipoprotein lipase

Glycerol + ATP ----->

Glycerol-3-P + ADP | Enzyme: Glycerol kinease

Glycerol-3-P -------->

Dihydroxyacetone-P | Enzyme: GPO

In the end, the produced H2O2 is converted to... by what enzyme... and what is produced?

Water by Peroxydase (POD) (Using 4-aminoantipyrine+phenol color = forming substr) RED QUINONE DERIVATES is produced.

What are the important materials when measuringTriglyceride in BP?

``` BP Colour reagent: - Enzymes - Colourforming substrate (4-aminoantipyrine + phenol) disolved in phosphate buffer. Standard triglyceride solution. Spectrometer ```

Equation to measure the Triclyceride conc. in blood plasma?

Equation: Abs sample --------------------- *2,28 = c sample Abs standard

What is produced during degradation of cholesterol in the liver?

Primary BILE ACIDS (cholic ac, chenocholic ac) are produced.

The salt of the primary bile acids with glycine and taurine gets excreted in the bile, and gets into the small intestines. What is the effect?

They have a detergent effect, so they play a role in DIGESTION and ABSORPTION of LIPIDS

Which primary bile acids do we determinate in the lab?

Taurocholic acid and | Glycocholic acid

What is the principal of the reaction of the primary bile acids?

Sucrose is turned into HYDROXYMETHYL FURFURAL by concentrated SULPHURIC ACID. OXYMETHYL FURFURAL produces a coloured complex with cholic acid.

What is the name of the reaction when determining cholic acid from the bile?

Pettenkofer´s reaction

What is the reactions of Pettenkofer´s reaction?

H2SO4 Sucrose ------------> Hydroxymethyl furfural Oxymethyl furfural + cholic acid -----> Coloured complex

What are the important materials in the Petterkofers reaction?

BILE (replaced with CHOLIC ACID) SUCROSE solution (10%) Conc. SULPHURIC ACID

Where is the enormous energy gain of the LACTATION of DAIRY COWS mainly covered?

By the LIPID MOBILIZATION from the fat depots | - LIPOLYSIS is INCREASED in the ADIPOSE TISSUE

LIPID MOBILIZATION from the fat depots goes like?

Lipolysis increased in adipose tissue Free fatty acids are released into the blood stream Free FA is mainly taken up by the liver In the liver they enter the BETA OXIDATION

Additional to the Lipolysis there is also a very intensive reaction happening in the hepatocytes at the same time? What does it produce?

GLUCONEOGENESIS - Producing a huge amount of GLUCOSE for LACTOSE Synthesis. - Oxalloacetate also enters mainly the GNG

How/why does the GNG turn into KETOGENESIS?

In the case of GNG, the high amount of acetyl CoA, produced in the BETA OXIDATION of FA, cannot enter the CITRIC ACID CYCLE because the LACK OF OXALOACETATE, they enter the KETOGENESIS

Cause of Ketogenesis, what does the hepatocytes produce?

Hepatocytes produces a high amount of KETONE BODIES (acetone, acetoacetate, beta-hydroxybutyrate= - Released in to the blood stream - Can be utilized by the extra hepatic tissue in the KETOLYSIS

Where is Ketone bodies produced, and where does it excrete?

In the liver Because GNG only happens in the liver Extretes mainly in urine, then milk

What happens in the beginning and peak of the lactation in dairy cows?

The ketogenesis is so intensive that the whole amount of the produced KETONE BODIES cannot be broken down in the KETOLYSIS, therefore: KB accumulate in the blood (Hyperketonaemia) and will be excreted with URINE (Ketonuria) and the Milk (Ketolactia) = Disease of ketosis develops.

What does Ketone bodies consist of?

Acetate Acetoacetate beta-hydroxybutyrate

Detection of KB has a huge diagnostic importance in dairy cow pracsis, but why?

Kidney concentrate the KB = The urine high am. of them | KB only exerted in the milk in SMALLER CONC.

What are the primary bile acids and where are they produced?

Cholic acid and Chenocholic acid IN LIVER

What are the secondary bile acids and where are they produced?

Taurocholic acid Glycocholic acid IN SMALL INTESTINES

Sodium notroprusside forms which color with 2 chemical compounds if which solution is present?

Sodium notroprusside forms PURPLE color with ACETOACETATE and ACETONE if AMONIA is present

What is the background of the detection of KB in the Urin and Milk?

Sodium nitroprusside Ammonia + ----------------> Purple colour complex Acetoacetate & Acetone

What is the proportion of KB conc. in diff biological fluids?

MILK:BLOOD:URINE | 1 : 5 : 10

What does it mean if the KB are present in the milk?

Severe KETOSIS has allready developed

How will Semiquantitative detection of KB be carried out?

From urine and milk samples from a dairy cow at the peak of lactation.

What are the important material then detecting the KB in Urine and Milk?

``` Ross reagent: - SODIUM NITROPRUSSIDE solution - Saturated with AMMONIUM SULPHATE 25% AMMONIA SOLUTION URINE and MILK ```

What does Ross reagent consist of?

SODIUM NITROPRUSSIDE and | AMONIUM SULPHATE

Lab 10 - Examination of lipids Flashcards

(70 cards)