BIOMAX Coomb’s Reagent
Biomax Rheumatoid Factor (RF) Slide Latex Kit
Biomax Widal Antigen Test Kit
Biotech: ALAT / ALT / GPT Reagents — Clinical Chemistry / Semi-Automated Systems-biochemistry
🔬 1. IFCC Kinetic Method (Standard ALT Method)
Primary Reaction
ALT catalyzes:
L-Alanine + α-Ketoglutarate
→ Pyruvate + L-Glutamate
Indicator (Coupled) Reaction
Pyruvate + NADH + H⁺
→ Lactate + NAD⁺
(catalyzed by LDH)
As ALT activity increases, NADH decreases, and absorbance at 340 nm falls.
Measurement
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Wavelength: 340 nm
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Method: Kinetic, ΔA/min
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Rate proportional to ALT activity (U/L)
Biotech: Albumin Reagents- biochemistry
🔬 1. BCG Method (Bromocresol Green)
Most widely used in routine chemistry analyzers.
Principle
Albumin binds to BCG dye in an acidic buffer forming a green-colored complex.
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Absorbance: 600–630 nm (green complex)
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Type: Endpoint reaction (color intensity ∝ albumin concentration)
Advantages
-
Very robust
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Strong signal (high sensitivity)
Limitations
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Mild positive bias in some conditions (e.g., high globulins)
🧪 2. BCP Method (Bromocresol Purple)
A more specific albumin method.
Principle
Albumin binds selectively to BCP dye in acidic buffer forming a purple complex.
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Absorbance: 540–550 nm
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Type: Endpoint
Advantages
-
Higher specificity than BCG
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Less interference from globulins or acute phase proteins
Limitations
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Slightly lower color intensity
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More common in premium reagent lines
Biotech: Alkaline Phosphatase (ALP / ALKP) Reagents biochemistry
🔬 1. IFCC Kinetic ALP Method (Standard)
Principle
ALP + p-Nitrophenyl phosphate (pNPP)
→ p-Nitrophenol (yellow) + Phosphate
-
Absorbance increases as p-nitrophenol is formed.
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The rate of increase (ΔA/min) at 405 nm is proportional to ALP activity.
Measurement
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Wavelength: 405 nm
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Mode: Kinetic (multiple interval readings)
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Temperature: 37°C (IFCC standard)
🧪 2. Reagent Composition (Typical)
Most ALP kits use single-reagent or two-reagent formats.
R1 (Main Buffer Reagent)
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Diethanolamine (DEA) buffer (high pH ~10.3)
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Magnesium ions (Mg²⁺) — ALP cofactor
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Zinc ions (Zn²⁺) — enzyme stabilizer
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Surfactants
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Preservatives
R2 (Substrate Reagent)
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p-Nitrophenyl phosphate (pNPP)
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Stabilizers
-
Preservatives
Biotech: ASAT / AST / SGOT Reagents — Clinical Chemistry / Semi-Automated Systems-biochemistry
🔬 1. IFCC Kinetic Method (Standard AST Method)
Primary Reaction (AST enzyme reaction)
L-Aspartate + α-Ketoglutarate
→ Oxaloacetate + L-Glutamate
Indicator (Coupled) Reaction
Oxaloacetate + NADH + H⁺
→ Malate + NAD⁺
(catalyzed by MDH)
LDH is often included to convert pyruvate → lactate, preventing inhibition.
As AST activity increases, NADH decreases, causing a fall in absorbance at 340 nm.
Measurement
-
Wavelength: 340 nm
-
Mode: Kinetic (rate), ΔA/min
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Rate proportional to AST activity (U/L)
Biotech: Biochemistry Reagents
1. Diagnostic Purpose
Biochemistry reagents are designed to quantitatively measure analytes such as:
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Liver function markers (ALT, AST, ALP, GGT, Bilirubin, Total Protein)
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Renal function markers (Urea, Creatinine, Uric Acid)
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Electrolytes (Sodium, Potassium, Chloride—depending on analyzer type)
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Lipid profile (Cholesterol, Triglycerides, HDL, LDL)
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Glucose metabolism (Glucose, HbA1c kits depending on system)
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Enzymes and substrates (LDH, CK, Amylase, Lipase)
2. Types of Formulations
Biochemistry reagents may come in:
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Single-reagent systems
(One ready-to-use bottle) -
Two-reagent systems
(R1 buffer + R2 starter/working reagent) -
Lyophilized reagents
(Require reconstitution) -
Liquid-stable reagents
(Long shelf-life, preferred for automated analyzers)
3. Compatibility
Designed for:
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Fully automated clinical chemistry analyzers
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Semi-automated bench-top analyzers
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Point-of-care analyzers (specific formats)
Biotech: Calcium Reagents-biochemistry
Principle of Measurement
1. Arsenazo III Method (Most widely used in modern analyzers)
How it works:
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Calcium ions react with Arsenazo III dye to form a blue-purple complex.
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The intensity of the color is measured photometrically at 650–660 nm.
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Highly specific for calcium with minimal interference from magnesium.
Advantages:
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Excellent stability
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Strong absorbance
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High specificity and precision
2. o-Cresolphthalein Complexone (OCPC) Method
How it works:
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Calcium forms a purple-red complex with o-cresolphthalein complexone in alkaline conditions.
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Read photometrically at 570–580 nm.
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Common in semi-auto chemistry analyzers.
Advantages:
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Reliable and widely established
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Suitable for manual and automated systems
Reagent Components
Arsenazo III Reagents May Contain:
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Arsenazo III dye
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Buffer (MES or other suitable buffers)
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Stabilizers
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Surfactants
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Preservatives
OCPC Reagents May Contain:
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o-Cresolphthalein complexone
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Alkaline buffer (e.g., 8-hydroxyquinoline to remove magnesium interference)
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Dye stabilizer
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Surfactants
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Preservatives
Packaging Format
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Single liquid reagent (most common)
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Two-reagent system (R1 + R2)
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Ready-to-use liquid stable bottles
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Analyzer-specific reagent cartridges
Common volumes: 25 ml, 50 ml, 100 ml, 250 ml or more.
Sample Types
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Serum
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Plasma (heparinized samples preferred)
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Urine (dilution may be needed)
Biotech: Chloride Reagents-biochemistry
Measurement Methods & Reagents
1. Ion-Selective Electrode (ISE) Method (Most common in modern analyzers)
Reagents include:
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Reference (internal standard) solution
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Electrode electrolyte solution
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Conditioning solution
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Cleaning solution
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Calibrators (Low and High Chloride)
Principle:
A chloride-selective membrane develops an electrical potential proportional to chloride concentration.
ISE systems require stable calibration solutions and maintenance reagents.
2. Mercuric Thiocyanate Colorimetric Method
How it works:
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Chloride reacts with mercuric ions, forming mercuric chloride.
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Released thiocyanate ions form a red-colored complex with ferric ions.
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The color intensity is measured photometrically at 480–520 nm.
Reagents contain:
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Mercuric thiocyanate
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Ferric nitrate or ferric ammonium sulfate
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Buffer system
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Surfactants and stabilizers
Advantages:
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Reliable for automated chemistry analyzers
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High precision
3. Colorimetric TPTZ Method (Less common)
Chloride displaces iron from a complex; the released iron reacts with TPTZ dye to form a measurable color.
Packaging Formats
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Single-liquid reagent
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Two-reagent kits (R1 buffer + R2 color reagent)
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ISE-specific reagent packs
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Ready-to-use liquid stable formulations
Typical volumes: 25 ml, 50 ml, 100 ml, 250 ml.
Biotech: Creatinine Reagents
1. Jaffe Method (Most Common)
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Based on the reaction between creatinine and picric acid in an alkaline medium.
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Produces an orange-red complex.
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Absorbance is measured photometrically, usually at 505–520 nm.
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Some kits include compensated Jaffe formulations to reduce interference from glucose, ketones, and proteins.
2. Enzymatic Method
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Uses a sequence of enzyme reactions (creatininase, creatinase, sarcosine oxidase, peroxidase).
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Produces a colored dye measurable at 550–570 nm.
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Higher specificity than Jaffe, minimal interference.
Reagent Components
Depending on the method, creatinine reagent kits may include:
For Jaffe Method
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Alkaline buffer
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Picric acid
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Stabilizers
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Surfactants
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Preservatives
For Enzymatic Method
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Enzyme mixture (creatininase, creatinase, sarcosine oxidase)
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Chromogenic dye reagents
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Phosphate or Good’s buffer
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Activators
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Preservatives
Packaging Format
Most creatinine reagent kits are provided as:
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Two-reagent system (R1 buffer + R2 working reagent)
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Ready-to-use liquid formulations
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Optional standards or calibrators
Common sizes: 25 ml, 50 ml, 100 ml, 250 ml, or analyzer-specific cartridges.
Sample Type
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Serum
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Plasma (heparin, EDTA)
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Urine (often diluted before analysis)
Biotech: GGT / Gamma GT Reagent-biochemistry
Principle of the Test
Most GGT reagents use the Szasz kinetic method, which involves:
-
Substrate:
γ-glutamyl-p-nitroanilide
This compound contains a gamma-glutamyl group. -
Enzymatic Reaction:
GGT in the sample transfers the gamma-glutamyl group to an acceptor (often glycylglycine). -
Product Formation:
The reaction releases p-nitroaniline, which has a strong yellow color. -
Measurement:
The analyzer measures the increase in absorbance at 405 nm.
The rate of color formation is directly proportional to the GGT activity.
Key Reagent Components
The GGT reagent typically contains:
✔ γ-glutamyl-p-nitroanilide
The chromogenic substrate used in the Szasz method.
✔ Glycylglycine (or similar acceptor)
Accepts the gamma-glutamyl group during the reaction.
✔ Buffers
Maintain optimal pH (usually alkaline, around pH 8.2–8.4).
✔ Stabilizers & Preservatives
Ensure reagent stability and long shelf life.
✔ Detergents / Surfactants
Help maintain reagent clarity and enhance reaction performance.
