1Which reagent is commonly used in the Dubois method for carbohydrate estimation?
Determination of carbohydrates by Dubois method
Easy
A.Ninhydrin and acetone
B.Benedict's reagent
C.Phenol and sulfuric acid
D.Biuret and sodium hydroxide
Correct Answer: Phenol and sulfuric acid
Explanation:
The Dubois method uses phenol and concentrated sulfuric acid to develop a colored product from carbohydrates.
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2What is the main purpose of concentrated sulfuric acid in the Dubois method?
Determination of carbohydrates by Dubois method
Easy
A.To hydrolyze carbohydrates
B.To precipitate proteins
C.To reduce glucose
D.To bind oxygen
Correct Answer: To hydrolyze carbohydrates
Explanation:
Concentrated sulfuric acid hydrolyzes and dehydrates carbohydrates, allowing them to react with phenol and produce color.
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3Which instrument is used to measure the color intensity in the Dubois method?
Determination of carbohydrates by Dubois method
Easy
A.pH meter
B.Colorimeter
C.Centrifuge
D.Water bath
Correct Answer: Colorimeter
Explanation:
A colorimeter measures the absorbance of the colored solution produced in the Dubois reaction.
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4The color developed in the Dubois method is generally measured at approximately which wavelength?
Determination of carbohydrates by Dubois method
Easy
A.
B.
C.
D.
Correct Answer:
Explanation:
The carbohydrate-phenol reaction commonly shows maximum absorbance near .
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5Which substance is commonly used to prepare the carbohydrate standard curve?
Determination of carbohydrates by Dubois method
Easy
A.Urea
B.Cholesterol
C.Albumin
D.Glucose
Correct Answer: Glucose
Explanation:
Glucose is commonly used as the standard for estimating the carbohydrate concentration.
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6In the Dubois method, a higher absorbance usually indicates a higher concentration of:
Determination of carbohydrates by Dubois method
Easy
A.Chloride ions
B.Carbohydrates
C.Lipids
D.Hemoglobin
Correct Answer: Carbohydrates
Explanation:
Within the working range, color intensity and absorbance increase as carbohydrate concentration increases.
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7What is the role of a reagent blank in the Dubois method?
Determination of carbohydrates by Dubois method
Easy
A.To increase sample concentration
B.To neutralize the sample
C.To provide a protein standard
D.To correct for reagent absorbance
Correct Answer: To correct for reagent absorbance
Explanation:
The blank contains the reagents without the analyte and accounts for absorbance due to the reagents themselves.
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8Which protein is commonly used as the standard in Lowry's method?
Determination of total proteins by Lowry's method
Easy
A.Glycine
B.Glucose
C.Bovine serum albumin
D.Cholesterol
Correct Answer: Bovine serum albumin
Explanation:
Bovine serum albumin, commonly called BSA, is widely used to prepare the protein standard curve.
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9Which reagent provides the biuret reaction in Lowry's method?
Determination of total proteins by Lowry's method
Easy
A.Glucose oxidase reagent
B.Alkaline copper reagent
C.Phenol-sulfuric acid reagent
D.Ninhydrin reagent
Correct Answer: Alkaline copper reagent
Explanation:
In alkaline conditions, copper ions react with peptide bonds to form the basis of the biuret reaction.
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10Which reagent is added in the second step of the Lowry method to increase color development?
Determination of total proteins by Lowry's method
Easy
A.Iodine solution
B.Folin-Ciocalteu reagent
C.Benedict's reagent
D.Molisch reagent
Correct Answer: Folin-Ciocalteu reagent
Explanation:
The Folin-Ciocalteu reagent reacts with protein-associated residues and intensifies the blue color.
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11What is measured in the Lowry method to estimate protein concentration?
Determination of total proteins by Lowry's method
Easy
A.Melting point
B.Flame temperature
C.Blue color intensity
D.Gas volume
Correct Answer: Blue color intensity
Explanation:
Protein produces a blue-colored complex in the Lowry reaction, and its intensity is measured spectrophotometrically.
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12The absorbance in the Lowry method is commonly measured near which wavelength?
Determination of total proteins by Lowry's method
Easy
A.
B.
C.
D.
Correct Answer:
Explanation:
The blue color produced by the Lowry reaction is commonly measured at approximately .
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13The Lowry method primarily estimates the concentration of:
Determination of total proteins by Lowry's method
Easy
A.Total protein
B.Blood glucose
C.Uric acid
D.Total carbohydrate
Correct Answer: Total protein
Explanation:
The Lowry method is a colorimetric assay used to determine the total protein concentration in a sample.
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14Why is a standard curve used in the Lowry method?
Determination of total proteins by Lowry's method
Easy
A.To sterilize the protein sample
B.To measure sample pH
C.To determine unknown protein concentration
D.To remove copper ions
Correct Answer: To determine unknown protein concentration
Explanation:
The absorbance of an unknown sample is compared with known protein standards to calculate its concentration.
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15Which enzyme is central to the glucose oxidase method?
Determination of blood glucose by glucose oxidase method
Easy
A.Pepsin
B.Amylase
C.Glucose oxidase
D.Urease
Correct Answer: Glucose oxidase
Explanation:
Glucose oxidase specifically catalyzes the oxidation of glucose in this method.
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16What product is formed when glucose is oxidized by glucose oxidase?
Determination of blood glucose by glucose oxidase method
Easy
A.Hydrogen peroxide
B.Ammonia
C.Urea
D.Lactic acid
Correct Answer: Hydrogen peroxide
Explanation:
Glucose oxidase converts glucose to gluconic acid and produces hydrogen peroxide as a reaction product.
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17Which enzyme commonly reacts with hydrogen peroxide in the color-producing step?
Determination of blood glucose by glucose oxidase method
Easy
A.Catalase
B.Peroxidase
C.Lipase
D.Trypsin
Correct Answer: Peroxidase
Explanation:
Peroxidase uses the hydrogen peroxide formed in the reaction to oxidize a chromogen and produce color.
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18In the glucose oxidase method, the intensity of the color is related to the concentration of:
Determination of blood glucose by glucose oxidase method
Easy
A.Blood calcium
B.Blood protein
C.Blood glucose
D.Blood urea
Correct Answer: Blood glucose
Explanation:
More glucose produces more hydrogen peroxide and therefore greater color intensity within the assay range.
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19Which instrument is commonly used to measure the colored product in the glucose oxidase method?
Determination of blood glucose by glucose oxidase method
Easy
A.Microscope
B.Centrifuge
C.Spectrophotometer
D.Analytical balance
Correct Answer: Spectrophotometer
Explanation:
A spectrophotometer measures the absorbance of the colored reaction product.
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20Why is a fasting blood sample often used for glucose estimation?
Determination of blood glucose by glucose oxidase method
Easy
A.To increase protein levels
B.To reduce dietary effects
C.To prevent enzyme activity
D.To remove all plasma water
Correct Answer: To reduce dietary effects
Explanation:
Fasting helps minimize the temporary increase in blood glucose caused by recent food intake.
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21What is the main purpose of adding concentrated sulfuric acid in the Dubois method?
Determination of carbohydrates by Dubois method
Medium
A.To oxidize carbohydrates into colored products
B.To reduce sugars into alcohols
C.To neutralize the phenol reagent
D.To precipitate carbohydrates from solution
Correct Answer: To oxidize carbohydrates into colored products
Explanation:
Concentrated sulfuric acid dehydrates carbohydrates to form furfural derivatives, which react with phenol to produce a measurable colored compound.
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22A carbohydrate sample gives an absorbance of 0.42. The standard curve indicates that an absorbance of 0.42 corresponds to 60 μg of glucose in the assay tube. If 0.5 mL of sample was used, what is the concentration of carbohydrate in the sample?
Determination of carbohydrates by Dubois method
Medium
A.120 μg/mL
B.240 μg/mL
C.30 μg/mL
D.60 μg/mL
Correct Answer: 120 μg/mL
Explanation:
The assay contains 60 μg in 0.5 mL, so the concentration is μg/mL.
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23Why is a reagent blank used in the Dubois carbohydrate assay?
Determination of carbohydrates by Dubois method
Medium
A.To correct for absorbance from reagents
B.To convert absorbance into fluorescence
C.To prevent carbohydrate hydrolysis
D.To increase the carbohydrate concentration
Correct Answer: To correct for absorbance from reagents
Explanation:
The blank contains all reagents except the carbohydrate sample and provides the background absorbance that must be subtracted.
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24A test sample produces an absorbance above the highest point of the glucose standard curve. What should be done before calculating its concentration?
Determination of carbohydrates by Dubois method
Medium
A.Increase the wavelength during measurement
B.Use the highest standard value directly
C.Subtract the sample absorbance from zero
D.Dilute the sample and repeat the assay
Correct Answer: Dilute the sample and repeat the assay
Explanation:
An absorbance outside the calibration range cannot be estimated reliably. Dilution brings the sample into the standard curve range.
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25A 1:5 dilution of a carbohydrate sample gives a calculated concentration of 18 μg/mL. What is the concentration in the original sample?
Determination of carbohydrates by Dubois method
Medium
A.3.6 μg/mL
B.18 μg/mL
C.90 μg/mL
D.23 μg/mL
Correct Answer: 90 μg/mL
Explanation:
The original concentration is obtained by multiplying the diluted concentration by the dilution factor: μg/mL.
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26In the Dubois method, why must the reaction mixture be mixed carefully after adding sulfuric acid?
Determination of carbohydrates by Dubois method
Medium
A.To prevent phenol from reacting
B.To remove all dissolved oxygen
C.To distribute heat and reagents uniformly
D.To decrease the final absorbance
Correct Answer: To distribute heat and reagents uniformly
Explanation:
The reaction with concentrated sulfuric acid is strongly exothermic. Proper mixing promotes uniform color development and improves reproducibility.
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27A standard curve for the Dubois assay is linear between 10 and 100 μg/mL. A diluted sample is estimated as 65 μg/mL, and the sample had been diluted fivefold. Which result should be reported for the original sample?
Determination of carbohydrates by Dubois method
Medium
A.65 μg/mL
B.325 μg/mL
C.500 μg/mL
D.13 μg/mL
Correct Answer: 325 μg/mL
Explanation:
The measured diluted concentration is multiplied by the dilution factor: μg/mL.
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28Which sequence best describes the chemical basis of the Lowry protein assay?
Determination of total proteins by Lowry's method
Medium
A.Biuret reaction followed by Folin reagent reduction
B.Benedict reaction followed by phenol reduction
C.Ninhydrin reaction followed by iodine oxidation
D.Sakaguchi reaction followed by sulfuric acid hydrolysis
Correct Answer: Biuret reaction followed by Folin reagent reduction
Explanation:
Peptide bonds first form a copper complex under alkaline conditions, and the Folin–Ciocalteu reagent is then reduced mainly by aromatic amino acid residues.
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29A protein sample has an absorbance corresponding to 40 μg of protein in the assay tube. If 0.2 mL of a 1:10 diluted sample was added, what was the protein concentration in the original sample?
Determination of total proteins by Lowry's method
Medium
A.4,000 μg/mL
B.2,000 μg/mL
C.200 μg/mL
D.20 μg/mL
Correct Answer: 2,000 μg/mL
Explanation:
The diluted sample contains μg/mL. Correcting for the tenfold dilution gives μg/mL.
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30Why is an appropriate protein standard, such as bovine serum albumin, used to construct the Lowry calibration curve?
Determination of total proteins by Lowry's method
Medium
A.To prevent alkaline hydrolysis of peptide bonds
B.To convert proteins into carbohydrate derivatives
C.To remove copper ions from the reaction mixture
D.To relate absorbance to an estimated protein concentration
Correct Answer: To relate absorbance to an estimated protein concentration
Explanation:
Known concentrations of the standard provide the relationship between absorbance and protein amount, allowing unknown samples to be quantified.
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31A protein extract contains a high concentration of detergent that produces an unexpectedly high absorbance in the Lowry assay. What is the most appropriate response?
Determination of total proteins by Lowry's method
Medium
A.Measure the sample without a reagent blank
B.Add more Folin reagent to cancel its effect
C.Replace the protein standard with glucose
D.Remove or dilute the detergent before analysis
Correct Answer: Remove or dilute the detergent before analysis
Explanation:
Some detergents interfere with color development or absorbance measurements. Removing or sufficiently diluting the interfering substance improves accuracy.
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32Why should the Folin–Ciocalteu reagent be added after the alkaline copper incubation in the Lowry assay?
Determination of total proteins by Lowry's method
Medium
A.The Folin reagent prevents peptide bond formation
B.The reagent must first digest all proteins
C.The copper–protein complex must form first
D.The alkaline reagent permanently removes copper
Correct Answer: The copper–protein complex must form first
Explanation:
The Lowry reaction depends on the initial biuret complex formation, followed by reduction of the Folin reagent to produce the blue color.
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33A sample gives an absorbance of 0.30, while the reagent blank gives 0.08. What absorbance should be used for interpolation from the protein standard curve?
Determination of total proteins by Lowry's method
Medium
A.0.30
B.0.08
C.0.22
D.0.38
Correct Answer: 0.22
Explanation:
The blank-corrected absorbance is calculated as .
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34If a protein sample has an absorbance above the linear range of the Lowry standard curve, why is direct extrapolation undesirable?
Determination of total proteins by Lowry's method
Medium
A.The wavelength will automatically change
B.The blank will become chemically inactive
C.The assay response may no longer be proportional
D.The protein will necessarily precipitate
Correct Answer: The assay response may no longer be proportional
Explanation:
Outside the linear range, absorbance may not increase proportionally with protein concentration, so dilution and reassay are preferred.
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35Which reaction product is directly generated when glucose oxidase acts on β-D-glucose?
Determination of blood glucose by glucose oxidase method
Medium
A.Carbon monoxide
B.Hydrogen peroxide
C.Urea
D.Lactic acid
Correct Answer: Hydrogen peroxide
Explanation:
Glucose oxidase converts glucose to gluconic acid or gluconolactone while producing hydrogen peroxide, which is used for color development.
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36In a glucose oxidase–peroxidase assay, the intensity of the colored product is primarily related to the amount of:
Determination of blood glucose by glucose oxidase method
Medium
A.Hydrogen peroxide formed
B.Oxygen initially dissolved
C.Protein precipitated
D.Chloride ions consumed
Correct Answer: Hydrogen peroxide formed
Explanation:
Peroxidase uses the hydrogen peroxide produced by glucose oxidase to oxidize a chromogen. More glucose generally produces more colored product.
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37A glucose standard has a concentration of 100 mg/dL and an absorbance of 0.50. An unknown sample has a blank-corrected absorbance of 0.35. Assuming linearity, what is the glucose concentration of the sample?
Determination of blood glucose by glucose oxidase method
Medium
A.100 mg/dL
B.70 mg/dL
C.35 mg/dL
D.143 mg/dL
Correct Answer: 70 mg/dL
Explanation:
Using proportionality, mg/dL.
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38Why should blood samples for glucose estimation be separated from cells promptly when analysis is delayed?
Determination of blood glucose by glucose oxidase method
Medium
A.Cells convert glucose into protein standards
B.Plasma immediately destroys glucose oxidase
C.Cells continue consuming glucose after collection
D.Separation increases the sample's oxygen content
Correct Answer: Cells continue consuming glucose after collection
Explanation:
Blood cells continue glycolysis in vitro, causing glucose concentration to fall if serum or plasma is not separated promptly or glycolysis is not inhibited.
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39A reducing substance in a blood sample consumes hydrogen peroxide during the glucose oxidase assay. What result is most likely obtained?
Determination of blood glucose by glucose oxidase method
Medium
A.A result independent of absorbance
B.No change in the glucose value
C.A falsely low glucose value
D.A falsely high glucose value
Correct Answer: A falsely low glucose value
Explanation:
Consumption of hydrogen peroxide leaves less peroxide for chromogen oxidation, reducing color intensity and causing underestimation of glucose.
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40A blood sample is diluted twofold before analysis and gives a calculated glucose concentration of 82 mg/dL from the diluted solution. What is the glucose concentration in the original sample?
Determination of blood glucose by glucose oxidase method
Medium
A.164 mg/dL
B.41 mg/dL
C.246 mg/dL
D.82 mg/dL
Correct Answer: 164 mg/dL
Explanation:
The original concentration is the measured diluted concentration multiplied by the dilution factor: mg/dL.
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41A carbohydrate standard curve is described by , where is in . A diluted unknown gives . If the sample was diluted tenfold before analysis, what is the carbohydrate concentration in the original sample?
Determination of carbohydrates by Dubois method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
Blank-corrected absorbance is . Thus, in the diluted sample. Multiplying by the dilution factor of 10 gives .
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42In the Dubois assay, the reagent blank has an absorbance of , whereas an unknown has an absorbance of . If the calibration equation based on blank-corrected readings is , what concentration should be used for the unknown?
Determination of carbohydrates by Dubois method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
The corrected absorbance is . Therefore, .
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43A sample gives absorbances of , , and after being diluted 2-, 4-, and 8-fold, respectively. The standard curve is linear only up to . Which result provides the most defensible estimate of the original concentration?
Determination of carbohydrates by Dubois method
Hard
A.Use the 2-fold dilution because it has the highest analyte concentration
B.Use the 4-fold dilution because its absorbance lies within the linear range
C.Average all three values because replicate dilutions are mathematically equivalent
D.Use the 8-fold dilution because dilution always improves analytical accuracy
Correct Answer: Use the 4-fold dilution because its absorbance lies within the linear range
Explanation:
The 8-fold dilution is outside the stated linear range, while the 2-fold dilution may be acceptable but has a higher risk of nonlinearity. The 4-fold dilution lies comfortably within the validated range and is therefore the best choice.
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44Two samples contain equal masses of glucose and starch, but the assay is calibrated with glucose. Which interpretation is most appropriate?
Determination of carbohydrates by Dubois method
Hard
A.They must produce identical absorbances because both are carbohydrates
B.Starch may produce a different response because color yield depends on sugar structure
C.Starch cannot react because only reducing sugars are detected
D.Glucose produces no response unless an oxidase is added
Correct Answer: Starch may produce a different response because color yield depends on sugar structure
Explanation:
The Dubois reaction measures acid-induced dehydration products, and different carbohydrates can have different color yields. Results are therefore conventionally expressed as equivalents of the calibration standard.
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45A carbohydrate assay is performed with concentrated sulfuric acid added to a phenol-containing mixture. The absorbance is unusually low in every tube, including standards, but the blank is normal. Which procedural error most plausibly explains this pattern?
Determination of carbohydrates by Dubois method
Hard
A.The cuvette was rinsed with distilled water
B.The acid was not mixed rapidly and uniformly after addition
C.The standards were incubated slightly longer
D.The sample dilution factor was recorded incorrectly
Correct Answer: The acid was not mixed rapidly and uniformly after addition
Explanation:
The reaction depends strongly on rapid, reproducible acid mixing and heat development. Inadequate mixing can cause incomplete or inconsistent color formation across standards and samples.
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46A diluted unknown gives a measured absorbance of . Its matrix-matched sample blank is , while the reagent blank is . If the calibration curve uses reagent-blank-corrected values, which absorbance should be used for quantification?
Determination of carbohydrates by Dubois method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
The matrix contribution must be removed in addition to the reagent background: .
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47A sample analyzed in triplicate gives concentrations of , , and , but a spiked aliquot expected to increase the concentration by gives . What is the best conclusion?
Determination of carbohydrates by Dubois method
Hard
A.The assay has quantitative recovery and no matrix effect
B.The triplicate precision proves that the result is accurate
C.The standard concentration was necessarily prepared incorrectly
D.The sample contains an inhibitor causing incomplete color development
Correct Answer: The sample contains an inhibitor causing incomplete color development
Explanation:
The observed increase is only instead of the expected , indicating poor recovery. A matrix effect such as inhibition or suppression is likely despite good replicate precision.
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48A Lowry calibration equation is , with in . A diluted sample gives an absorbance of and was prepared by diluting of serum to . What is the protein concentration in the original serum?
Determination of total proteins by Lowry's method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
The diluted concentration is . The dilution factor is , so the original concentration is . Since the options express the result in the stated assay scale, the correct value is .
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49A Lowry assay gives for an unknown and for the reagent blank. The standard curve was generated using blank-corrected absorbances with slope . What is the concentration of the measured solution?
Determination of total proteins by Lowry's method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
The blank-corrected absorbance is . Dividing by the slope gives .
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50A protein extract contains sodium dodecyl sulfate, and its Lowry result is much higher than expected. Which explanation is most scientifically justified?
Determination of total proteins by Lowry's method
Hard
A.SDS has no possible effect because the Lowry reaction measures only nitrogen
B.SDS can interfere with color development or protein precipitation in the assay
C.SDS is a protein and is therefore measured stoichiometrically as albumin
D.SDS destroys all peptide bonds, making the signal falsely quantitative
Correct Answer: SDS can interfere with color development or protein precipitation in the assay
Explanation:
The Lowry assay is sensitive to matrix composition. Detergents such as SDS can alter protein behavior and the Folin reaction, producing biased results unless the method is validated for that matrix.
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51Why can two proteins at the same mass concentration produce different Lowry absorbances when bovine serum albumin is used as the standard?
Determination of total proteins by Lowry's method
Hard
A.The method measures molecular mass directly rather than chemical reactivity
B.All proteins have identical color yields, but cuvettes alter protein structure
C.The Lowry method detects only carbohydrate groups attached to proteins
D.The Lowry response depends partly on amino-acid composition and aromatic residues
Correct Answer: The Lowry response depends partly on amino-acid composition and aromatic residues
Explanation:
Lowry color formation includes copper complexation and reduction of Folin reagent, with substantial contribution from residues such as tyrosine and tryptophan. Results are therefore protein-standard equivalents.
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52A set of Lowry standards is linear through but becomes curved above that concentration. An unknown gives a reading corresponding to on the curved portion. What is the best next action?
Determination of total proteins by Lowry's method
Hard
A.Report because the instrument displayed a numerical value
B.Subtract the highest standard absorbance from the unknown absorbance
C.Dilute the unknown until its reading falls within the validated linear range
D.Extrapolate the linear segment to estimate the unknown concentration
Correct Answer: Dilute the unknown until its reading falls within the validated linear range
Explanation:
Quantification should use interpolation within the validated linear range. Diluting the unknown and correcting for the dilution factor avoids unreliable extrapolation from the curved region.
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53Which change would most directly compromise comparability among Lowry tubes even if all tubes contain the correct protein concentrations?
Determination of total proteins by Lowry's method
Hard
A.Using matched cuvettes with identical optical path lengths
B.Adding Folin reagent at different times and reading tubes after unequal intervals
C.Including a reagent blank containing no protein
D.Preparing all standards from the same protein stock solution
Correct Answer: Adding Folin reagent at different times and reading tubes after unequal intervals
Explanation:
The Folin color-development step is time-dependent. Unequal reaction times produce different absorbances unrelated to protein concentration, so synchronized timing is essential.
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54A protein sample contains a high concentration of ammonium sulfate after precipitation. The measured Lowry protein concentration is substantially elevated. Which control is most useful for diagnosing this bias?
Determination of total proteins by Lowry's method
Hard
A.Measure the sample without adding the alkaline copper reagent
B.Analyze a protein-free matrix containing the same ammonium sulfate concentration
C.Increase the wavelength from to
D.Replace all protein standards with glucose standards
Correct Answer: Analyze a protein-free matrix containing the same ammonium sulfate concentration
Explanation:
A matrix-matched blank containing ammonium sulfate but no protein reveals whether the salt contributes to absorbance or alters color development. This distinguishes chemical interference from true protein signal.
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55In the glucose oxidase-peroxidase method, which sequence best represents the analytical chemistry?
Determination of blood glucose by glucose oxidase method
Hard
A.Glucose binds peroxidase directly, and oxygen is released as the colored product
B.Glucose is converted to lactate, which reduces ferricyanide to a colored product
C.Glucose reduces the chromogen directly, and catalase generates the final dye
D.Glucose produces hydrogen peroxide, which supports peroxidase-mediated chromogen oxidation
Glucose oxidase converts glucose and oxygen to gluconic acid and hydrogen peroxide. Peroxidase then uses hydrogen peroxide to oxidize a chromogen, producing the measured color.
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56A blood sample contains a high concentration of ascorbate. In a glucose oxidase-peroxidase color assay, what bias is most likely if no interference correction is used?
Determination of blood glucose by glucose oxidase method
Hard
A.No bias because glucose oxidase oxidizes every reducing substance equally
B.Falsely low glucose because ascorbate consumes hydrogen peroxide or reduces the dye
C.Falsely high glucose because ascorbate is converted directly into glucose
D.Random variation only because ascorbate changes the cuvette path length
Correct Answer: Falsely low glucose because ascorbate consumes hydrogen peroxide or reduces the dye
Explanation:
Ascorbate can scavenge hydrogen peroxide and reduce the oxidized chromogen. Both effects decrease color formation and can produce a falsely low glucose result.
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57Whole blood is left at room temperature before glucose analysis. The glucose result is lower than expected, although the assay reagents and calibration are valid. What is the most likely cause?
Determination of blood glucose by glucose oxidase method
Hard
A.Glucose oxidase converted glucose into protein during storage
B.Room temperature prevented all glucose from entering the plasma
C.Continued glycolysis by blood cells consumed glucose before separation
D.Hemoglobin spontaneously increased the glucose concentration
Correct Answer: Continued glycolysis by blood cells consumed glucose before separation
Explanation:
Erythrocytes and leukocytes continue metabolizing glucose after collection. Prompt plasma or serum separation, cooling, or an appropriate antiglycolytic preservative helps prevent this decrease.
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58A glucose assay has a standard absorbance of for and an unknown absorbance of . The sample was diluted 1:5 before testing. Assuming a linear zero-intercept response, what is the original glucose concentration?
Determination of blood glucose by glucose oxidase method
Hard
A.
B.
C.
D.
Correct Answer:
Explanation:
The diluted sample concentration is . Correcting for the fivefold dilution gives .
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59Two glucose samples have identical glucose concentrations, but one is markedly hemolyzed. In a peroxidase-based colorimetric glucose oxidase assay, which issue should be considered first?
Determination of blood glucose by glucose oxidase method
Hard
A.Hemoglobin permanently inhibits glucose transport into the reaction mixture
B.Hemoglobin is converted quantitatively into glucose by glucose oxidase
C.Hemoglobin may contribute optical interference near the measurement wavelength
D.Hemolysis eliminates oxygen and therefore makes all glucose undetectable
Correct Answer: Hemoglobin may contribute optical interference near the measurement wavelength
Explanation:
Hemoglobin absorbs strongly in portions of the visible spectrum and can alter apparent absorbance. Hemolyzed samples should be assessed for spectral or chemical interference under the specific assay conditions.
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60A patient sample gives a glucose value above the highest calibrator. Which response best preserves quantitative validity?
Determination of blood glucose by glucose oxidase method
Hard
A.Use the reagent blank as the new highest calibrator
B.Report the value by extrapolating the calibration line
C.Repeat the assay after diluting the sample into the calibration range
D.Multiply the absorbance by the highest calibrator concentration
Correct Answer: Repeat the assay after diluting the sample into the calibration range
Explanation:
A result above the calibration range is not reliably quantified by extrapolation. Dilution followed by multiplication by the dilution factor places the measurement within the validated response interval.
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