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Citric Acid (CA) Colorimetric Assay Kit– MSE Supplies LLC

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Citric Acid (CA) Colorimetric Assay Kit

SKU: E-BC-K351-M-96

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Citric Acid (CA) Colorimetric Assay Kit

SKU # E-BC-K351-M
Detection Instrument Microplate reader (535 nm-555 nm, optimum wavelength: 545 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms CA
Sample type Serum, plasma, animal tissue, mitochondria
Sensitivity 0.06 mmol/L
Detection range 0.06-2.0 mmol/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 50 min
Precision Average inter-assay CV: 4% | Average intra-assay CV: 4%
Other instruments required Micropipettor, Water bath, Incubator, Vortex mixer, Centrifuge
Storage 2-8℃
Valid period 12 months


Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.06-2.0 mmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Human serum 5-20
Dog serum 5-10
Rat serum 10-20
Horse serum 5-10
Mouse plasma 5-20
10% Rat brain tissue homogenate 5-10
10% Rat liver tissue homogenate 5-20
10% Rat kidney tissue homogenate 5-10
10% Rat lung tissue homogenate 15-30
10% Mouse heart tissue homogenate 5-20

 

Note: The diluent is reagent 1.

 

Detection Principle

In biochemistry, citric acid is an intermediate in the citric acid cycle and plays an important role in metabolism. Citric acid levels in blood and urine are affected by factors such as age, gender, diet, citric acid precursors, and parathyroid hormone and sex hormones.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Extracting Solution 60 mL × 1 vial 60 mL × 2 vials 2-8°C, 12 months
Reagent 2 Lysis Buffer 10 mL × 1 vial 20 mL × 1 vial 2-8°C, 12 months
Reagent 3 Reducing Agent Powder × 1 vial Powder × 1 vial 2-8°C, 12 months
shading light
Reagent 4 Chromogenic Agent 1.2 mL × 1 vial 1.2 mL × 2 vials 2-8°C, 12 months
shading light
Reagent 5 2 mmol/L CA Standard 2 mL × 1 vial 2 mL × 1 vial 2-8°C, 12 months
Microplate 96 wells No requirement
Plate Sealer 2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)

Parameters Sample 1 Sample 2 Sample 3
Mean (mmol/L) 0.55 1.02 1.80
%CV 4.3 3.9 3.8


Inter-assay Precision

Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (mmol/L) 0.55 1.02 1.80
%CV 3.9 4.2 3.9


Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 95%.


Standard 1 Standard 2 Standard 3
Expected Conc. (mmol/L) 0.45 0.85 1.3
Observed Conc. (mmol/L) 0.4 0.8 1.2
Recovery rate (%) 96 94 95

 

Sensitivity

The analytical sensitivity of the assay is 0.06 mmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.


Standard Curve

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (mmol/L) 0.0 0.2 0.5 0.8 1.0 1.2 1.5 2.0
OD value 0.071 0.091 0.112 0.136 0.147 0.154 0.177 0.204
0.071 0.091 0.106 0.127 0.142 0.156 0.172 0.205
Average OD 0.071 0.091 0.109 0.132 0.145 0.155 0.174 0.205
Absoluted OD  0.000 0.020 0.038 0.061 0.074 0.084 0.103 0.134