View Article

Abstract

The present work to Synthesize benzotriazole Nucleus and study the biological activity of Antimitotic method through by seed germination assay by using the green gram seeds. It is preliminary screening method to identify this benzotriazole basic nucleus is having Anticancer property. By using this seed germination assay method be can studied that this nucleus is having inhibition of seed germination Activity.

Keywords

Benzotriazole Synthesis, Antimitotic Method, Diazotisation Reaction, Seed Germination Assay.

Introduction

Benzotriazole is Bicyclic heterocyclic compound with three nitrogen atoms in the five membered ring i.e. triazole ring fused with benzene ring. This Pharmacophore molecule shows a wide Spectrum of pharmacological activities.

Table.1: Chemical Information of Benzotriazole [1-3]

 

Synonyms

1H-Benzotriazole, Benzotriazole,1,2,3-Benzotriazole

Chemical formula

C6H5N3

Molecular weight

119.12 g/mole

Chemical structure

        <a href="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-4.jpg" target="_blank">
            <img alt="1.jpg" height="150" src="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-4.jpg" width="150">
        </a>
 

Density

1.36 g/ml

Melting point

100?c

Appearance

White solid

Chemical safety category

Environmental Hazard, Harmful

Table.2: Materials used for the Reaction

Apparatus

Chemicals

Weighing balance

orthophenylene diamine

Glassine

Sodium Nitrite

Ice water bath

Glacial acetic acid

Thermometer

Distilled water

RFB

 

Beaker

Measuring cylinder

Heating mantle

Gravity filtration setup

Desiccator

Principle involved in the synthesis of Benzotriazole: [4-6]

The sodium nitrite reacts with glacial acetic acid and liberates nitrous acid.The o-phenylenediamine reacts with nitrous acid and produce diazonium ion. When the structure and stereochemistry of diazonium ion are stable, intramolecular nitrogen coupling occurs and forms benzotriazole.

NaNO2   + CH3COOH                      HNO2 + CH3COONa

        <a href="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-3.png" target="_blank">
            <img alt="2.png" height="150" src="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-3.png" width="150">
        </a>
Table.3: Procedure for the Preparation of the Reaction mixture:

Reaction mixture-1

Reaction mixture-2

Dissolve1.3grams of O-phenylenediamine in a mixture of 1.5ml of glacial acetic acid and 5ml water in a beaker. Stir until the solid dissolves, warm gently if necessary. Cool the solution to 15?c.Stir well

Add a solution of 2grams of sodium nitrite in 2ml water.

Mixing RM-1 & RM-2 then this resultant reaction mixture will become warm within2-3minutes and reaches a temperature of about 85?c. and then begin to cool. Colour changes from deep red to pale brown. Continue stirring for 15minutes till the temperature fall about 35-40?c.Thoroughly chill in the ice bath for 30minutes.Filter the product and wash with cold water. Product is dried under Desiccator.

Evaluation of Antimitotic activity of Benzotriazole compound [7-12]

Table.4: Materials used for seed germination Assay method

Contents

Quantity

Green grams (Vigna radiata)

25grams

Sigma-Aldrich Polystyrene petri dishes

5 petri dishes

Benzotriazole drug solution (BZT)

50ml BZT

Vehicle used-Distilled water

50ml

Seed germination Assay Method:

Green gram seeds were purchased from the market and stored in a moisture free place in self-sealing cover. Then accurately weighed 5grams of seeds and incorporated in each and every sigma-Aldrich polystyrene petri dishes to study the antimitotic activity of Benzotriazole compound. In Control only vehicle is used that vehicle is distilled water and incubate all the petridishes at room temperature for 24hrs. After getting the results of antimitotic activity of Benzotriazole compound. we randomly selected 3 seeds from each and every petriplate and calculate the germination length in cms of the seeds and after that we done average length  calculation.

Preparation of Benzotriazole drug solution:

Accurately weight about 1gm of Benzotriazole drug from the synthesized product and dissolved in the distilled water then make up to 50ml with the distilled water in the Volumetric flask.

        <a href="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-2.png" target="_blank">
            <img alt="BZT Drug Solution.png" height="150" src="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-2.png" width="150">
        </a>
Figure.1: BZT Drug Solution             

Figure.2: Green Gram Seeds

Table.5: Preparation of Antimitotic activity of Benzotriazole (BZT) compound.

S.no

Concentration of Benzotriazole compound (ml)

Contents in the petriplate

1

Control (0ml BZT)

10ml vehicle only

2

0.5ml (BZT)

10ml vehicle + 0.5ml BZT

3

1ml (BZT)

10mlvehicle +1ml BZT

4

1.5ml (BZT)

10mlvehicle + 1.5ml BZT

5

2ml (BZT)

10mlvehicle + 2ml BZT

        <a href="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-1.png" target="_blank">
            <img alt="Results of Antimitotic Assay method for Benzotriazole Drug solution.png" height="150" src="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-1.png" width="150">
        </a>
Figure.3: Results of Antimitotic Assay method for Benzotriazole Drug solution

Table.6: Result of Antimitotic Activity of Benzotriazole compound.

 

S. No

Concentration of Benzotriazole compound (ML)

Length of seeds (cm)

Average length of the seeds(cm)

1

 

Control

First Seed- 4cm

Second seed -4.5cm

Third seed -4.5cm

 

4+4.5+4.5= 13/3

=4.3cms

2

 

0.5ml

First Seed- 2.5cm

Second seed-3cm

Third seed -3.7cm

 

2.5+3+3.7 = 9.2/3

= 3.06cms

3

 

1ml

First seed -1.5cm

Second seed-2cm

Third seed-2.5cm

 

1.5+2+2.5= 6/3

=2cms

4

1.5ml

First seed -0.9cm

Second seed -0.5cm

Third seed -0cm

 

0.9+0.5+0 = 1.4/3

= 0.46cms

5

2ml

First seed -0cm

Second seed -0cm

Third seed -0cm

 

No germination of seeds

From the Figure.3 We obtained the results of antimitotic activity from this we calculated the average length of seeds germination from each and every individual concentrations of Benzotriazole (BZT) drug solution.Then after we plot a graph for taking the Average length of seeds in Y-xis and concentration of Benzotriazole drug solution in X-Axis.

        <a href="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-0.png" target="_blank">
            <img alt="Graphical Representation of Antimitotic Assay of Benzotriazole Drug Solution.png" height="150" src="https://www.ijpsjournal.com/uploads/createUrl/createUrl-20250424120805-0.png" width="150">
        </a>
Figure.4: Graphical Representation of Antimitotic Assay of Benzotriazole Drug Solution

CONCLUSION:

From the Above results of Antimitotic activity of Benzotriazole (BZT) Drug solution we observe that there is a gradually reduction in Average length of the seeds.so we conclude that highest strength of the BZT drug solution is hot having any seed germination from this it is conformed that the Benzotriazole Compound is having Antimitotic activity.

REFERENCES

  1. Navneet Singh, Vanika Priya, Ridhima Bhasin, Kunika Verma. An exploration of sundry nature of benzotriazole derivatives: A Review. International research journal of pharmacy. 2019; 10 (3)
  2. Suma B.V., Natesh N.N., Madhavan V. Benzotriazole in medicinal chemistry: an overview. J. Chem. Pharm. Res. 2011;3:375–381
  3. John H.B., John M.B.J. eleventh ed. 2004. Wilson and Gisvold's Textbook of Organic Medicinal and Pharmaceutical Chemistry; pp. 217–281.
  4. Christopher John Perry, Keith Holding and Elizabeth Tyrrell. (2008). “Simple, Novel Synthesis for 1CarbamoyllH-benzotriazole and Some of Its derivatives ” Synthetic Communications: An International Journal for Rapid Communication of Synthetic Organic Chemistry.
  5. Henderson S.A., Yang B. Applications of benzotriazole methodology in heterocycle ring synthesis and substituent introduction and modification. J. Heterocyclic. Chem. 1998;35:1123–1159.
  6. Zhang S.S., Wan J., Li C.L., Li M., Qu B. Syntheses and biological activities of novel triazole compounds. J. Heterocyclic. Chem. 2007;44:75–78.
  7. V K Singh, Poonam Rish Ishwar, Peeyush Bhardwaj and Shashi Alok. (2017). Kale R., Prasad V., Mohapatra P., Tiwari V. Recent developments in benzotriazole methodology for construction of pharmacologically important heterocyclic skeletons. Monstah fur Chem. - Chem. Mon. 2010;141:1159–1182.
  8. Gitanjali KP, Harshada CP, Indira MP, Prof. SL Borse and Dr. SP Pawar. (2015). “Benzotriazole – The Molecule Of Diverse Biological Activities” WJPPS, 4(5).
  9. V K Singh, Poonam Rish Ishwar, Peeyush Bhardwaj and Shashi Alok. (2017). Benzotriazole: A Heterocyclic Molecule with Diversified Pharmacological Activities, IJPSR, 8(2), 446-456
  10. Perkovic I, Butala I, Kralj M, Martin Kleiner I and Balzarini J.(2012). Novel NSAID 1-acyl 4cycloalkyl/aryl semi carbazides and 1-acyl-5-benzyloxy/hydroxyl carbamoyl carbazides as potential anticancer agents and antioxidants. Eur J Med Chem, 51, 227-238.
  11. Al-Soud Y.A., Al-Masoudi N.A., Ferwana Ael R. Synthesis and properties of new substituted 1,2,4-triazoles: potential antitumor agents. Bioorganic. Med. Chem. 2003;11:1701–1708
  12. Walck JL, Baskin JM, Baskin CC, Hid Ayati SN. Defining transient and persistent seed banks in species with pronounced seasonal dormancy and germination patterns. Seed Sci Res. 2005;15(3):189–196.

Reference

  1. Navneet Singh, Vanika Priya, Ridhima Bhasin, Kunika Verma. An exploration of sundry nature of benzotriazole derivatives: A Review. International research journal of pharmacy. 2019; 10 (3)
  2. Suma B.V., Natesh N.N., Madhavan V. Benzotriazole in medicinal chemistry: an overview. J. Chem. Pharm. Res. 2011;3:375–381
  3. John H.B., John M.B.J. eleventh ed. 2004. Wilson and Gisvold's Textbook of Organic Medicinal and Pharmaceutical Chemistry; pp. 217–281.
  4. Christopher John Perry, Keith Holding and Elizabeth Tyrrell. (2008). “Simple, Novel Synthesis for 1CarbamoyllH-benzotriazole and Some of Its derivatives ” Synthetic Communications: An International Journal for Rapid Communication of Synthetic Organic Chemistry.
  5. Henderson S.A., Yang B. Applications of benzotriazole methodology in heterocycle ring synthesis and substituent introduction and modification. J. Heterocyclic. Chem. 1998;35:1123–1159.
  6. Zhang S.S., Wan J., Li C.L., Li M., Qu B. Syntheses and biological activities of novel triazole compounds. J. Heterocyclic. Chem. 2007;44:75–78.
  7. V K Singh, Poonam Rish Ishwar, Peeyush Bhardwaj and Shashi Alok. (2017). Kale R., Prasad V., Mohapatra P., Tiwari V. Recent developments in benzotriazole methodology for construction of pharmacologically important heterocyclic skeletons. Monstah fur Chem. - Chem. Mon. 2010;141:1159–1182.
  8. Gitanjali KP, Harshada CP, Indira MP, Prof. SL Borse and Dr. SP Pawar. (2015). “Benzotriazole – The Molecule Of Diverse Biological Activities” WJPPS, 4(5).
  9. V K Singh, Poonam Rish Ishwar, Peeyush Bhardwaj and Shashi Alok. (2017). Benzotriazole: A Heterocyclic Molecule with Diversified Pharmacological Activities, IJPSR, 8(2), 446-456
  10. Perkovic I, Butala I, Kralj M, Martin Kleiner I and Balzarini J.(2012). Novel NSAID 1-acyl 4cycloalkyl/aryl semi carbazides and 1-acyl-5-benzyloxy/hydroxyl carbamoyl carbazides as potential anticancer agents and antioxidants. Eur J Med Chem, 51, 227-238.
  11. Al-Soud Y.A., Al-Masoudi N.A., Ferwana Ael R. Synthesis and properties of new substituted 1,2,4-triazoles: potential antitumor agents. Bioorganic. Med. Chem. 2003;11:1701–1708
  12. Walck JL, Baskin JM, Baskin CC, Hid Ayati SN. Defining transient and persistent seed banks in species with pronounced seasonal dormancy and germination patterns. Seed Sci Res. 2005;15(3):189–196.

Photo
Satish Ponnada
Corresponding author

Sri Sivani college of pharmacy, Andhra Pradesh.

Photo
Bhagyalaxmi Suvvari
Co-author

Sri Sivani college of pharmacy, Andhra Pradesh.

Satish Ponnada* Bhagyalaxmi Suvvari, Synthesis and Antimitotic Activity of Benzotriazole Compound, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 4, 2932-2937 https://doi.org/10.5281/zenodo.15273474

More related articles
Formulation and Evaluation of Nano emulsion-Based ...
Mohammad Azhar, Dr. Amul Mishra, ...
Bis-Chalcones: A Review On Synthetic Methodologies...
Auti Vivek Babasaheb , Swagati A Moon , ...
An Anti- Aging Herbal Soap of Centella Asiatica in Combination with Portulaca Ol...
Vaishnavi Wadnere, Prachi Murkute, Ashwini Pundkar, Santosh Payghan , Vaishnavi Sonune, ...
Evaluation of Phytochemicals in crude extracts derived from the Aerial parts of ...
Arnab Roy, Sajid Ansari, Subham Kumar Lohani , Ayush Kumar, Makhmur Hayat, ...
Related Articles
Study Of Adverse Drug Reaction Of Paracetamol Dose...
Atharv S. Gangarde, Kiran H. Bibave, ...
Formulation And Comparative Evaluation of Marketed Paracetamol Suspension ...
Kiran Pathade, Varsha Bhati, Sakshi Kshirsagar , Disha Gangule, Vaibhavi Jape, ...