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Magnesium serves as a cofactor for numerous enzymes and is present in both soft tissues and bone, with roughly equal distribution between the two. Its metabolic mechanisms are not yet fully understood. Elevated magnesium levels can lead to reduced muscle tone, while decreased levels are observed in conditions such as hypoparathyroidism and chronic renal failure.
Detection Principle
In an alkaline solution, magnesium ions bind with the Calmagite reagent to form a magenta-red complex. The color intensity is proportional to the magnesium ion concentration. The absorbance is measured at 510 nm using a spectrophotometer, and the magnesium content is calculated based on a formula. The solution contains the calcium chelator EGTA to eliminate calcium interference. The inclusion of a surfactant stabilizes the protein colloid, allowing for direct magnesium measurement in serum without protein removal. This kit is for research use only and is not suitable for clinical diagnosis or other purposes.
| M1506748 | Component | 100T | Storage |
| M1506748A | Magnesium Standard (0.823 mmol/L) | 1 mL | 2-8℃ |
| M1506748B | Calmagite Chromogenic Solution | 25 mL | 2-8℃. Store in the dark. |
| M1506748C | Mg Assay Buffer | 25 mL | 2-8℃. Store in the dark. |
| M1506748D | Calmagite Base Solution | 2.5 mL | 2-8℃ |
Materials Required but Not Provided
Deionized water, dilute potassium hydroxide solution, or dilute hydrochloric acid solution
Centrifuge tubes or test tubes, centrifuge, cuvettes, spectrophotometer
Experimental Procedure
1. Sample Preparation
1.1 Plasma/Serum Samples
Plasma or serum samples prepared by standard methods can be used directly. Store at -20°C for Mg detection.
1.2 Cell or Tissue Samples
Homogenize an appropriate amount of cells or tissue. Centrifuge at low speed to collect the supernatant. Store at -20°C for Mg detection.
1.3 High Concentration Samples
If the sample contains a high Mg concentration, dilute with ddH₂O. The use of ordinary distilled water is not recommended.
Note: After preparation, determine the protein concentration using a BCA protein assay kit to calculate the Mg content per unit protein weight in tissues or cells. The Aladdin B665595 BCA Protein Quantification Kit or R1491648 Ready-to-Use BCA Protein Quantification Kit is recommended.
2. Preparation of Mg Chromogenic Working Solution
Immediately before use, mix the components in the ratio: Calmagite Chromogenic Solution : Mg Assay Buffer : Calmagite Base Solution : Deionized Water = 10 : 10 : 1 : 80. Adjust the pH to 11.5 ± 0.2 using 1M potassium hydroxide solution. This is the Mg Chromogenic Working Solution. Store at 4°C protected from light; stable for 2 weeks.
3. Mg Assay Setup
Use dry test tubes cleaned with dilute hydrochloric acid and rinsed with deionized water, or disposable sterile polyethylene centrifuge tubes. Set up Blank, Standard, and Test tubes according to the table below. Add reagents in the specified order, avoiding bubbles. For high Mg concentrations, reduce the sample volume or dilute appropriately. Running duplicates is recommended.
| Reagent (mL) | Blank Tube | Standard Tube | Test Tube |
| ddH₂O | 0.04 | — | — |
| Magnesium Standard (0.823 mmol/L) | — | 0.04 | — |
| Test Sample | — | — | 0.04 |
| Mg Chromogenic Working Solution | 2 | 2 | 2 |
4. Mg Measurement
Mix well. Using the Blank tube to zero the instrument, measure the absorbance of the Standard and Test tubes at 510 nm using a cuvette with a 1 cm light path (recorded as Astandard and Atest).
5. Calculation of Results
Serum/Plasma Magnesium:
Magnesium (mmol/L) = (Atest / Astandard) × 0.823
Tissue Magnesium:
Magnesium (mmol/mg prot) = (A<sub>test</sub> / A<sub>standard</sub>) × 0.823 / Protein concentration of test sample (mg/L)
Parameter Description:
Atest: Absorbance of the test tube
Astandard: Absorbance of the standard tube
Unit Conversion: mg/dL = mmol/L / 0.411
Reference Interval: Serum magnesium concentration in healthy adults: 0.7 ~ 1.1 mmol/L
Precautions
Hemolyzed samples interfere with the assay; avoid their use whenever possible.
Use reagents promptly after opening to prevent potential effects on subsequent experiments.
Lipemic samples also interfere; delipidate samples before assay.
Under this kit's conditions, the magnesium ion concentration in the test sample should ideally be greater than 0.05 mmol/L and less than 0.823 mmol/L. For concentrations above 0.6 mmol/L, dilute with deionized water before measurement to avoid potential errors.
This method can be adapted for automated biochemical analyzers using an endpoint detection method.
If the sample concentration is too high, dilute with ddH₂O and re-assay, multiplying the result by the dilution factor.
Avoid contamination with Mg²⁺ ions, as this can affect the detection results.
For your safety and health, wear a lab coat and disposable gloves during operation.
Comprehensive hazard, handling, storage, and regulatory compliance document.
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