Carbon monoxide hemoglobin detection kit (colorimetric method)

Cat. No.: C1507809

GRADE & PURITY BioReagent

Overview

Carbon monoxide poisoning occurs when the incomplete combustion of carbon‑containing materials produces carbon monoxide, which is then inhaled through the respiratory tract. The mechanism of poisoning is that carbon monoxide has an affinity for hemoglobin hundreds of times greater than that of oxygen, allowing it to bind readily with hemoglobin to form carboxyhemoglobin (HbCO). This binding deprives hemoglobin of its oxygen‑carrying capacity and function, leading to tissue hypoxia. Carbon monoxide exerts toxic effects on all tissue cells, with the most severe impact on the cerebral cortex.

Detection Principle:

After carbon monoxide binds to hemoglobin, it forms cherry‑red carboxyhemoglobin (HbCO). HbCO is not reduced by sulfite, whereas oxyhemoglobin (HbO₂) is reduced. HbCO has an absorption peak at 538 nm, and HbO₂ has an absorption peak at 578 nm. The ratio of the two absorbances is used in a formula to calculate the HbCO content. This method is primarily used for the quantitative detection of carboxyhemoglobin (carbon monoxide hemoglobin) in human and animal blood. This kit is intended for research use only and is not suitable for clinical diagnosis or other purposes.

Reagents, consumables and Equipments not provided

1. Distilled water

2. Cuvettes or 96‑well plate, spectrophotometer or microplate reader

Procedure

1.. Dilute the Alkaline Buffer (10×) to 1× with distilled water. Store tightly sealed and set aside for use.

2. Accurately weigh several portions of Sulfite, each portion being 20 mg. Set aside for use.

3. Take several large test tubes and add 0.1 mL of fresh blood to each. Also take 0.1 mL of blood from a healthy non‑smoker as a control. Add 20 mL of 1× Alkaline Buffer to each tube and mix well.

4. Immediately add 20 mg of Sulfite to each tube and mix thoroughly.

5. Transfer an appropriate amount of the above blood mixture into cuvettes or a 96‑well plate. Measure the absorbance at 538 nm (A₅₃₈) and at 578 nm (A₅₇₈) using a spectrophotometer or microplate reader. 

Note: Measurements must be completed within 10 minutes.

6. Calculation of Results

HbCO (%) = {2.44 × (A₅₃₈ / A₅₇₈) – 2.68} × 100%

Reference Interval:

Population	HbCO Level
Normal individuals	0.2%–0.5%
Urban residents	0.5%–1.5%

Notes

1. Each assay should include a normal control.

2. Avoid contamination of the Alkaline Buffer with acids or alkalis, and store it tightly sealed.

3. For your safety and health, please wear a lab coat and disposable gloves during operation.

4. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.

Storage and Shipping

Storage

Room temperature

Shipped In

Normal

Stability And Storage

Store at room temperature long term (12 months).

Contents & Storage

C1507809	Component	50T	Storage
C1507809A	Alkaline Buffer (10×)	100 mL	RT.
C1507809B	Sulfite	2 g	RT.