Apollo Sci. 1,3,5-Triphenyltetrazolium formazan

Apollo Scientific 1,3,5-Triphenyltetrazolium Formazan (TPF) is a high-purity biochemical product extensively used as a critical analytical reference and colorimetric standard. As the insoluble red reduction product of 2,3,5-Triphenyltetrazolium Chloride (TTC), TPF serves primarily in biochemical, microbiological, agricultural, and environmental research, facilitating accurate quantitative analyses of enzyme activity, cell viability, and metabolic activity.

Chemical Identity and Specifications

• Chemical Name: 1,3,5-Triphenyltetrazolium Formazan
• Common Abbreviation: TPF
• CAS Number: 548-73-2
• Molecular Formula: C19H16N4
• Molecular Weight: 300.36 g/mol
• Appearance: Dark red, crystalline solid
• Solubility: Insoluble in water; soluble in organic solvents such as ethanol, methanol, acetone, and DMSO.

Principle and Mechanism of Action

TPF is the stable, insoluble red-colored formazan product generated through enzymatic or chemical reduction of the colorless tetrazolium salt TTC. It serves primarily as a standard reference compound in assays involving TTC reduction. The formation and quantification of TPF directly correlate with cellular respiratory activity, metabolic enzyme function, microbial viability, or tissue health.

Applications and Detailed Use Cases

1. Analytical Reference for Cell Viability and Metabolic Assays

Primary Use:

• Serving as an analytical standard and calibration reference in colorimetric assays employing TTC.

Practical Example:

• Researchers performing TTC-based cell viability assays use pure TPF as a calibration standard to construct standard curves. This approach enables precise spectrophotometric quantification of metabolic activity in cell cultures, ensuring accurate and reproducible results.

2. Microbial Respiration and Enumeration Standards

Primary Use:

• Quantitative standard for microbial respiration assays that utilize TTC reduction, ensuring reliable and comparable results across experiments.

Practical Example:

• Microbiologists testing bacterial respiration and metabolic activity use Apollo Scientific TPF as a known concentration standard. By comparing absorbance values of experimental samples to TPF standards, precise microbial quantification and enumeration are possible in clinical, food safety, and environmental testing.

3. Seed Viability and Germination Studies (Agricultural Research)

Primary Use:

• Calibration and validation of seed viability assays based on TTC staining methods.

Practical Example:

• Agricultural scientists assessing viability of crop seeds use TPF standards to calibrate spectrophotometric measurements, ensuring that the intensity of TTC reduction accurately corresponds to seed metabolic activity. This method aids in accurate determination of seed quality and viability.

4. Tissue Viability and Histological Studies (Biomedical Research)

Primary Use:

• Reference standard in histological and pathological analyses involving TTC staining, especially in ischemia and infarction studies.

Practical Example:

• Researchers studying myocardial infarction or tissue ischemia employ TPF as a reference standard, verifying that staining intensity in tissue sections accurately reflects metabolic viability. This reference ensures accurate differentiation between healthy and necrotic tissues in pathological assessments.

5. Environmental Monitoring and Soil Microbial Activity

Primary Use:

• Quantitative standard and reference in environmental microbial activity assays utilizing TTC as a respiratory indicator.

Practical Example:

• Environmental microbiologists studying soil microbial respiration compare soil-derived TPF against pure TPF reference standards, accurately quantifying microbial activity, ecosystem health, and biodegradation efficiency in contaminated or remediated sites.

Advantages and Key Benefits

• Highly Pure Reference Compound: Ensures precise quantification, calibration, and reproducibility in biochemical assays.
• Stable and Consistent: Provides consistent and reliable calibration standards across multiple assay types.
• Versatile Application: Widely applicable as a reference standard in microbiological, biomedical, agricultural, and environmental studies.
• Easy-to-Use Standardization: Facilitates simple calibration of spectrophotometric and colorimetric assays involving TTC reduction.

Storage, Stability, and Handling

• Store tightly sealed in a cool, dry place (recommended storage: refrigerated at 2–8°C).
• Protect from direct exposure to light and moisture to maintain reagent integrity.
• Observe standard laboratory safety precautions for chemical handling.

Intended Users

• Research microbiology laboratories
• Biotechnology and pharmaceutical researchers
• Agricultural and seed-testing laboratories
• Biomedical and clinical pathology laboratories
• Environmental and ecological research facilities
• Academic institutions conducting biochemical and metabolic research

Apollo Scientific 1,3,5-Triphenyltetrazolium Formazan (TPF) serves as a critical reference and calibration reagent, supporting accurate quantification, reproducibility, and validity of essential biochemical and biological assays across multiple research disciplines.