Hemoglobin Content Assay Kit (SDS-Hb, Colorimetric Method)

Cat. No.: H1522004

GRADE & PURITY BioReagent

Vue d’ensemble

Hemoglobin (Hb or HGB) is a protein responsible for oxygen transport in higher organisms and gives blood its red color. Hemoglobin consists of four chains: two α-chains and two β-chains, each containing a cyclic heme group with one iron atom. Hb readily binds oxygen in oxygen‑rich environments and releases it in oxygen‑poor environments. This property enables red blood cells to transport oxygen. 

Current methods for hemoglobin detection include: cyanmethemoglobin method, alkaline hydroxyhemoglobin method, sodium dodecyl sulfate (SDS/SLS) binding method, copper sulfate titration method, or measurement with imported large‑scale biochemical analyzers. The cyanmethemoglobin method involves toxic potassium cyanide and hazardous waste. The copper sulfate titration method has large preparation errors and is sensitive to ambient temperature. Biochemical analyzers are expensive with high testing costs. 

Principle: This Hemoglobin Content Assay Kit (SDS-Hb, Colorimetric Method) works as follows: all forms of hemoglobin in blood except sulfhemoglobin (SHb) react with sodium dodecyl sulfate (SDS or SLS) to form a brown SLS‑Hb compound. The absorbance at 538 nm is proportional to the hemoglobin concentration, allowing quantification based on absorbance (A).

For the quantitative measurement of hemoglobin content in blood samples to assist in evaluating anemia, blood loss, etc. For research use only (RUO). Not for clinical diagnosis or other applications.

Kit Components:

H1522004	Components	Appearance	100T	Storage
H1522004A	Hb standard	Solid	20mg	2-8℃. Store in the dark.
H1522004B	SLS Stock Solution (100×)	Liquid	3mL	2-8℃.

Materials Required but Not Provided

1. Deionized or distilled water, physiological saline
2. EDTA anticoagulant tubes, centrifuge tubes, centrifuge, cuvettes, spectrophotometer

Procedure (For Reference Only)

1. Preheat the spectrophotometer for at least 30 min; set wavelength to 538–540 nm.

2. Use freshly collected anticoagulated blood directly. Hemolysate, serum, or plasma may also be used directly. Centrifuge turbid serum/plasma and store supernatant at 4°C until use. Dilute high‑Hb samples 2–5 fold with distilled water or saline.

3. Hb Standard Preparation: Dissolve 20 mg Hb Standard in 1 mL distilled water to make a 20 mg/mL (20 g/L) standard solution. Stable for 2 weeks at 2–8°C.

4. SLS Working Solution: Mix 1 volume of SLS Stock Solution (100×) with 99 volumes of deionized water.

5. Pipetting Scheme (Unit: mL)

Component	Blank	Standard	Test
Deionized water	0.05	—	—
Hb Standard (20 g/L)	—	0.05	—
Sample	—	—	0.05
SLS Working Solution	2.5	2.5	2.5

Mix thoroughly and incubate at room temperature for 5 min.

6. MeasurementAt 540 nm with a 1.0 cm cuvette, measure absorbance of blank (A₍blank₎), standard (A₍std₎), and test (A₍test₎) samples.

Calculation

Hemoglobin Concentration (g/L) = [(A₍test₎ − A₍blank₎) / (A₍std₎ − A₍blank₎)] × C₍std₎ × N Where:

• C₍std₎ = Hb concentration of standard = 20 g/L
• N = sample dilution factor

Precautions

1. The Hb Standard is not calibrated with HiCN; minor deviations may exist. Users may prepare certified standards if required.
2. Use fresh samples whenever possible. Store samples at 4°C if not assayed immediately.
3. Avoid contamination of the standard; store sealed at 4°C.
4. Linear range: 0–200 g/L. Dilute samples exceeding the upper limit with saline and multiply results by dilution factor.
5. Wear lab coats and disposable gloves for safety.
6. Use reagents promptly after opening to avoid performance decline.

Storage & Stability

Validity: 6 months. Transport under cold conditions; store at 4℃. 

Specifications

Spécifications et pureté

BioReagent

Grade

BioReagent

Application

Cell Metabolism

Type d'échantillon

Méthode de détection

Colorimetric Method

Detection instrument

UV-Visible Spectrophotometer

Detection wavelength

538nm

Sensibilité

Light-sensitive

Stockage et expédition

Conditions de stockage de stockage

Store at 2-8°C,Protected from light

Expédié en

Wet ice

Stabilité et stockage

Store at 2-8℃ long term (6 months). Store in the dark.

Contents & Storage

H1522004	Components	Appearance	100T	Storage
H1522004A	Hb standard	Solid	20mg	2-8℃. Store in the dark.
H1522004B	SLS Stock Solution (100×)	Liquid	3mL	2-8℃.