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Object: To Estimate Blood Sugar Level From Given Sample of Blood

This document summarizes the orthotoludine and enzymatic methods for determining blood sugar level from a blood sample. The enzymatic method involves using glucose oxidase and peroxidase to catalyze a reaction between glucose, phenol, and 4-aminophenazone to form a red-violet quinoneimine dye whose intensity is proportional to the glucose level. The test requires reagents, pipettes, test tubes, a spectrophotometer, blood sample, and thermostat equipment. Whole blood, serum, or plasma can be used. Results are calculated by comparing readings of the test and standard solutions and interpreting the glucose level.

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Yasir Mushtaq
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62 views2 pages

Object: To Estimate Blood Sugar Level From Given Sample of Blood

This document summarizes the orthotoludine and enzymatic methods for determining blood sugar level from a blood sample. The enzymatic method involves using glucose oxidase and peroxidase to catalyze a reaction between glucose, phenol, and 4-aminophenazone to form a red-violet quinoneimine dye whose intensity is proportional to the glucose level. The test requires reagents, pipettes, test tubes, a spectrophotometer, blood sample, and thermostat equipment. Whole blood, serum, or plasma can be used. Results are calculated by comparing readings of the test and standard solutions and interpreting the glucose level.

Uploaded by

Yasir Mushtaq
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOC, PDF, TXT or read online on Scribd
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Object: To Estimate blood Sugar level from given sample of blood.

Methods of determination 1. Orthotoludine method 2. Enzyemetic ethod PRINCIPLE:

Glucose is determined after enzymetic oxidation in the presence of glucose oxidase !he formed "ydrogen peroxidase reacts under catalysis of peroxidase #ith phenol and ! " #minophenazone red$violate %uinoneimine dye as indicator !he intensity of colour is directly proportional to the presence of sugar in $lood sample
RE%&IREMEN!'

Glucose enzymatic (it) measuring pippttes) test tu$es) 'pectrophotometer) $lood sample a$out * cc) !hermostat e+uipment Cotton s#a$) sprit) ,isposa$le 'yringes
'PECIMEN

-hole $lood) 'erum and Plasma


C.N!EN!'

/ Enzymatic reagent consist phosphate $uffer 01 23) 456minophenazone) Phenol) Glucose oxidase) Peroxidase) and Mutarotase 'ta$ilizers * 'tandard 'olution contain Glucose /77 mg8dl or 2 22 mmol8l
PR.CE,&RE

6''69 -a:elenght: .ptical path: !emperature: Measurement:

277 nm to 24; nm / cm *7< *2 7C or =1 7C 6gainst reagent $lan( .nly one reagent $lan( per series is re+uired

' No
/ * =

'olution
Enzymatic 'olution 'tandard 'erum

!est
* ml 5 7 7* ml

'ample
* ml 7 7* ml 5

>lan(
* ml 5 5

'ha(e #ell) incu$ate for *7 to *2 7C or 2 minutes at =1 7C and record the readings of a$sor$ance in test and standard separately $y 'pectrophotometer under #a:e length of 277 5 24; nm
C6LC&L6!I.N Reading of !est ? /77 @ Reading of 'tandard AAAAAAAAAAAAmg8dl

Result:5 !he gi:en sample of $lood contain 55555555555555555mg8dl Glucose 0Random 8 Basting3
IN!ERPRE!6!I.N: Clinical Importance

Normal Calue Basting @ ;7 to //7 mg8dl Random @ /*7 to /;7 mg8dl 'ource Car$ohydrate) Li:er Glycogen "9PERGL9CEMI6 0Increased $lood 'ugar le:el a$o:e /*7 mg 8dl3

/ ,ia$etes Mellitus * "yperacti:ity of thyroid) 0"perthyroidsim3) pituitary and adrenal gland 0accelerate a$sorption) insulin resistant) gluconeogenesis3 due to increased >MR = "aemochromatosis: Excessi:e Iron leads restriction D Cells 4 'tressful situations: Increased catecholamine and stimulate sympathetic system #hich suppress insulin 2 Pheochromocytoma 5555555555 0!umor of 6drenal Gland3: leads release catecholamine and glycogenolysis 0$rea(do#n of glycogen to glucose3 decreases release of insulin ; Pregnancy increased estrogen #hich is ,ia$etogenic hormone
"9P.GL9CEMI6 0,ecreased Glucose Le:el >elo# Normal Range 3 .:er dosage of Insulin Insulin 'ecreting !umors of Pancreas "ypothyroidism 0Myxoedema) Critinism3 decreased >MR "ypopitutarism decreased >MR "ypoadrenalism 06ddisonEs ,isease3 decreased >MR Certain Glycogen 'torage ,isease eg Con F Gier(Es disease 'e:ere Li:er ,isease) enzymatic deficiency Gluconeogenesis not occurs 6lcohol Ingestion formation of lactic acid 'e:ere exercise formation of lactic acid

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