Nutrition And Metamorphosis
Lab 3 and 4: Determination of Glucose, Cholesterol and Triglycerides in Whole Blood Using Reflotron
The usage of enzyme systems to observe and obtain quantitative measuring of analyte concentration is good established. Assorted enzymes systems have been stabilized and incorporated into dry reagent format with the necessary co-factors, coloring material reagents, buffer and so on, needed to bring forth a compact, accurate and disposable analytical component. Example of instrument includes the Reflotron system. The Reflotron® Plus Chemistry Analyzer is an in vitro diagnostic device designed for the quantitive finding of clinical chemical science parametric quantities utilizing Reflotron® Test reagent strips. It works on the rule of coefficient of reflection photometry and ensures rapid and dependable consequences within two to three proceedingss while being easy to utilize. The consequences are shown on a clear LC show and the profile can be printed out via the built-in pressman. The Reflotron® Plus possesses a storage capacity for 60 consequences with extra information. The Reflotron® contraption can besides be really easy linked to an external computing machine. This allows further processing of informations every bit good as informations exchange.
The Reflotron® Test are reagent strips designed for the specific quantitative finding of 17 clinical parametric quantities for the most common indicants utilizing undiluted specimen stuffs. The incorporation of a plasma-separating system makes it possible to find informations straight from capillary or venous whole blood every bit good as serum and plasma of little volume. The Reflotron® Test reagent strips consist of three basic constituents: a separation tablet, a reagent tablet, and a magnetic codification. The separation tablet is constructed so that plasma is separated from whole blood via glass fiber paper. The design of the tablet forces the detached plasma to flux to the reagent tablet, where the chemical reaction takes topographic point. The magnetic codification which located on the bottom of each reagent strip contain all test- and lot-specific informations so that no standardization is necessary by the user and allows for the transportation of up to 500 spots of electronic informations. ( Linda Dailey, 2006 )
Reflotron® Plus is designed for easy-to-use handling and low care. Its little dimensions take up really small infinite. The good thought-out characteristics make the finding of clinical-chemical parametric quantities quick, convenient and easy because there is no demand to fix reagent and standardization is besides unneeded. The immediate trial consequences guarantee a speedy public presentation or confirmation of the diagnosing without hold. The trial consequences can so be discussed instantly with the patient and any intervention or therapy can be done rapidly. The most of import indicants in primary attention are covered for such wellness jobs as diabetes, lipid upsets, kidney diseases, musculus diseases, anaemia, liver diseases, pancreatitis, urarthritis, and bone upsets. In add-on, the patient ‘s cardiac hazard harmonizing to “Framingham” or “PROCAM” , the Friedewald LDL cholesterin, the entire cholesterol/HDL cholesterin ratio, and the creatinine clearance can besides be calculated. ( Roche Diagnostic, neodymium )
Trial Principal of Reflotron® Plus
After application to the trial strip, the blood ( after separation of the erythricytes from the plasma ) flow into the reaction zone. D-glucose is oxidized to ?-D-gluconolactone by atmospheric O in the presence of glucose oxidized ( GOD ) . The ensuing H peroxide oxidizes an index in the presence of peroxidase ( POD ) . The dye formed in this mode is relative to the glucose concentration of the blood sample. ( Diagnostic in Clinical Chemistry I, nd )
Glucose + O2 > ?-D-gluconolactone + H2O2
H2O2 + Indicator > Dye + H2O
The Reflotron® Test cholesterin reagent strip contains cholesterol esterase, cholesterin oxidase, peroxidase and an index. Cholesterol esters are cleaved into cholesterin and the corresponding fatty acids by the catalytic action of cholesterin esterase. In the presence of O, the free cholesterin is oxidized to cholestenone with cholesterin oxidase moving as the accelerator. The H peroxide formed oxidizes the index to a bluish dye. The strength of the coloring material formed is straight relative to the cholesterin concentration in the whole blood. ( Linda Dailey, 2006 )
Triglycerides + 3 H2O > Glycerol + 3 RCOOH ( enzyme: esterase )
Glycerol + ATP > Glycerol-3-phosphate + ADP ( enzyme: glycerin kinase )
Glycerol-3-phosphate + O2 > dihydroxyacetone phosphate + H2O2 ( enzyme: GPO )
Indicator ( colourless ) + H2O2 > Indicator ( blue ) + H2O ( enzyme: Pod )
( Roche Diagnostic, neodymium )
Possible Test Mistake:
Damaged magnetic codification will do a “Code Error” show when inserted in the instrument.
Improper arrangement of sample ; unequal sum of sample may change trial consequences.
To find glucose, cholesterin and triglyceride concentration in whole blood utilizing Reflotron® Plus Chemistry Analyzer.
Reflotron® Plus Chemistry Analyzer
Reflotron® Capillary Pipettor and Pipette Tubes
Reflotron® Test reagent strips
1. The Reflotron® Plus Chemistry Analyzer was switched on. A “Ready” was displayed after the warm-up clip expired.
2. A Reflotron® Test reagent strip was taken out from vial. Make non flex and make non touch the black barcode on the strip.
3. The fingertip was swabbed by utilizing a unfertile swab and punctured by utilizing lancet device.
4. Blood sample was drawn by utilizing the Reflotron® Plus pipettor and applied onto the Centre of the xanthous application zone on the trial strip for glucose finding. Be careful non to touch the application zone with the pipette tip.
5. The trial strip was placed on the Reflotron® Plus Chemistry Analyzer with the magnetic side down and slides frontward horizontally until it was locked into topographic point. The skiding screen was so closed.
Note: as the strip was inserted, do certain that the magnetic codification was on the underside. If the show did non demo “Close Door” , the trial strip was removed and reinserted, doing certain that it was locked into topographic point. “Close Door” would be appeared on the show.
6. The consequence was shown on a clear LC show with the trial name and the concentration unit and the informations can be printed out via the built-in pressman.
7. The door was so opened ; the reagent strip was removed and discarded.
8. Reflotron® Plus Chemistry Analyzer was now ready for other trials of cholesterin and triglyceride.
Type of trial
Determination of 17 clinical-chemical parametric quantities from whole blood, serum or plasma.
Determination of glucose, cholesterin and triglyceride from whole blood, serum or plasma.
No standardization is necessary ( automatic standardization )
Lot specific via saloon codification
3 ten 1.5V Batteries
Immediate certification of consequences
No incorporate pressman
( Roche Diagnostic, neodymium )
Blood Glucose, Cholesterol and Triglyceride Levels
Type of Test
Time to Rest
What Value Means
8-16 hours fasting
Adult: 3.9-5.5 mmol/L
Pregnancy mark: 3.3-5.6 mmol/L
A degree of 5.6-6.9 mmol/L indicates impaired glucose tolerance/insulin opposition.
A fasting degree of 7.0 mmol/L or higher indicates type II diabetes.
Triglycerides ( TG )
8-16 hours fasting
& lt ; 1.7 mmol/L
Borderline-High: 1.7-2.3 mmol/L
High: 2.3-5.64 mmol/L
Very High: & gt ; 5.64 mmol/L
Elevated degrees are a hazard factor of coronary arteria disease and other upset including diabetes.
8-16 hours fasting
& lt ; 5.17 mmol/L
Borderline-High: 5.17-6.18 mmol/L
High: 6.21 mmol/L and over
Increased degrees are associated with addition hazard of bosom disease.
( MGH Department of Pathology, 2006 )
( A Drug-free Solution to Elevated Blood Triglycerides, neodymium )
( American Heart Association, 2009 )
Blood glucose changed instantly after a repast might be identified at finger sites before sensing at forearm or thigh sites. ( Ellison JM et Al, 2002 ) Therefore, in this experiment, blood was drawn from the fingertip of a female pupil ( 22year ) . In nondiabetic persons, fasting plasma glucose concentration ( following an nightlong 8- to 10-hours fast ) by and large range from 4.22 to 6.11 mmol/litre. Glucose concentrations began to lift ~10 min after a repast started as a consequence of the soaking up of dietetic saccharides. The magnitude and clip of the peak postprandial glucose concentration were depended on several factors, such as timing, measure, and composing of the repast. In normal persons, plasma glucose concentration extremum ~60 min after a repast started and returned to preprandial degree within 2-3 hours. ( American Diabetes Associaiton, 2001 )
The blood glucose degree of the tried pupil in this experiment was higher than normal scope, which was 8.11 mmol/L. This was because the pupil had eaten pieces of Tuna Sandwich 15minutes before the lab started and the blood was taken from the pupil ~30minutes after the lab had started. One big piece of white staff of life had glycemic index of 73 ( comparative to glucose ) and glycemic burden of 10. ( Jane Higdon, 2005 ) The high glycemix index of white staff of life would do spike in blood glucose degree. Determination of glucose was used for the diagnosing and monitoring of upsets of carbohydrate metamorphosis such as diabetes mellitus, idiopathic hypoglycemia and in pancreatic islet cell tumours. ( Diagnostic in Clinical Chemistry I, nd ) In general, a measuring of plasma glucose 2 hours after a repast started was practical. ( American Diabetes Associaiton, 2001 ) A normal glucose would be less than 7.8 mmol/L if taken approximately 2 hours after eating.
Triglycerides were taken up from nutrient or produced in liver or in adipose tissue, chiefly from saccharide. Triglycerides served as energy modesty in adipose tissue. Elevated triglyceride degrees were associated with a 30 % cardiovascular hazard addition in work forces and a 75 % addition in adult females. Increased triglyceride degrees were an independent hazard factor for cardiovascular diseases, normally associated with other lipid and nonlipid hazard factor such as high blood force per unit area and metabolic diseases such as diabetes and fleshiness. Non-fasting triglycerides may give a more accurate image of coronary bosom disease hazard than fasting triglycerides since triglyceride-rich lipoprotein are believed to be the most atherogenic. ( Claus Luley et Al, 2000 ) Blood triglycerides content would top out 3 hours after a high lipid repast ( Jill M. Slade et Al, 2007 ) The normal triglycerides range based on age of 10-29 old ages was 0.6-1.2 mmol/L. The postprandial blood triglycerides degree of tried pupil ( female, 22 twelvemonth ) was normal with the value of 0.963 mmol/L. However, the ideal degree for grownup male and female was 1.1mmol/L. ( A Drug-free Solution to Elevated Blood Triglycerides, neodymium )
Cholesterol was an of import steroid alcohol used in metamorphosis as a precursor of assorted steroid endocrines. 30 % -40 % of Cholesterol was taken up from nutrient and 60 % -70 % were produced in liver ( when kiping ) . Determination of entire cholesterin was indispensable for primary and secondary bar of cardiovascular diseases, since it was the premier hazard factor for lipoprotein metamorphosis upsets, coronary artery disease and coronary bosom disease. ( Linda Dailey, 2006 ) On norm, work forces develop coronary arteria disease ten to 15 old ages earlier than adult females do and their hazard for deceasing of bosom disease at younger ages than adult females is higher. Womans between the ages of 20 and 34 and after climacteric, around age 55, have higher cholesterin degrees than work forces do. Absorption of cholesterin was affected by the combination of nutrient in a repast. Some of the dietetic fibers constituent could adhere with cholesterin and be excreted in the fecal matters. The tried pupil had a normal postprandial blood cholesterin degree of 3.75 mmol/L.
The reagent strips were brought to room temperature before executing check because the strips were usually stored under 30?C.
All the reagent strips should be avoided from exposing to moisture by shuting the phial instantly after remotion of strips utilizing the original stopper which contains a drying agent.
The termination day of the month of the reagent strips were checked before usage.
A new pipette tip must be used for each sample.
All the reagent strips should be protected from magnetic Fieldss because the magnetic codification on the bottom may be damaged by magnetic Fieldss.
All sharps were placed in an appropriate biohazard sharps containers and follow acceptable wellness section disposal patterns.
The tried pupil had somewhat higher postprandial blood glucose degree of 8.11 mmol/L and normal postprandial blood cholesterin and triglycerides degrees with the values of 3.75 mmol/L and 0.963 mmol/L severally. Prevention was better than remedy. Therefore, we must supervise our blood glucose, triglyceride, and cholesterin degrees purely. We should pattern a healthy life manner by eating a heart-healthy diet, acquire regular physical activity and avoid baccy fume.
1. Ellison JM EL at, 2002, Rapid Changes in Postprandial Blood Glucose Produce Concentration Differences at Finger, Forearm, and Thigh Sampling Sites. Available from: & lt ; hypertext transfer protocol: //www.ncbi.nlm.nih.gov/pubmed/12032099 & gt ; [ Accessed: 21st September 2009 ]
2. Diagnostic In Clinical Chemistry I, nd Available from: & lt ; hypertext transfer protocol: //faculty.ksu.edu.sa/aabulhamd/Documents/III % 20lab/02- % 20Effect % 20of % 20specimen % 20handling % 20on % 20analytes.pdf & gt ; [ Accessed: 21st September 2009 ]
3. American Diabetes Associaiton, 2001, Postprandial Blood Glucose Available from: & lt ; hypertext transfer protocol: //care.diabetesjournals.org/content/24/4/775.full & gt ; [ Accessed: 21st September 2009 ]
4. Linda Dailey, 2006, Local Public Health Laboratories of Kentucky: Cholesterol by Reflotron Available from: & lt ; hypertext transfer protocol: //www.healthdepartment.org/LAB % 20Procedures % 20Manual/Cholesterol/Reflotron.pdf & gt ; [ Accessed: 21st September 2009 ]
5. Roche Diagnostic, nd, Reflotron® Plus. Available from: & lt ; hypertext transfer protocol: //www.diavant.com/diavant/CMSFront.html ; jsessionid=B55EBF502C22537F5C00AB5EEC073BF6? pgid=3,1,9,1 & gt ; [ Accessed: 21st September 2009 ]
6. American Heart Association, 2009, What your Cholesterol Levels Mean Available from: & lt ; hypertext transfer protocol: //www.americanheart.org/presenter.jhtml? identifier=183 & gt ; [ Accessed: 21st September 2009 ]
7. MGH Department of Pathology, 2006, MGH Clinical Laboratory Handbook Available from: & lt ; hypertext transfer protocol: //mghlabtest.partners.org/MGHRefRanges.pdf & gt ; [ Accessed: 21st September 2009 ]
8. Jane Higdon, 2005, Micronutrient Information Center: Glycemic Index and Glycemic Load. Available from: & lt ; hypertext transfer protocol: //lpi.oregonstate.edu/infocenter/foods/grains/gigl.html & gt ; [ Accessed: 21st September 2009 ]
9. Claus Luley et Al, 2000 Point-of-Care Testing of triglycerides: Evaluation of the Accutrend tridlycerides System. Available from: & lt ; hypertext transfer protocol: //www.clinchem.org/cgi/content/full/46/2/287 & gt ; [ Accessed: 21st September 2009 ]
10. Jill M. Slade et Al, 2007 The postprandial addition in blood triglycerides has no direct consequence on the encephalon BOLD response. Available from: & lt ; hypertext transfer protocol: //www3.interscience.wiley.com/journal/117910229/abstract? CRETRY=1 & A ; SRETRY=0 & gt ; [ Accessed: 21st September 2009 ]
11. A Drug-free Solution to Elevated Blood Triglycerides, nd, Fasting Blood Triglycerides Level: Current Guideline Available from: & lt ; hypertext transfer protocol: //www.reducetriglycerides.com/baseline_triglycerides_measurement.htm & gt ; [ Accessed: 21st September 2009 ]