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In this paper, a new series of substituted 1, 2,4-triazines was prepared by reacting 4-amino sulfanilic acid with substituted benzaldehyde to form the corresponding Schiff base compounds (1-5), and then reacting two moles of O-methoxy benzaldehyde with hydrazine to form a compound ( 6) and these compounds reacted with Schiff base (1-5) by applying the Diels-alder reaction in the presence of benzene as a solvent and refluxed it to cyclized into substituted of 1,2,4-triazines derivatives (7-11). Depending on the physical and spectroscopic properties of the prepared compounds that were identified, infrared spectroscopy and nuclear magnetic resonance were used. An evaluation of the biological activity of the prepared compounds was conducted and it gave anti-bacterial results, such as gram-positive and gram-negative when compared with known antibiotics.

This class of compounds, which is known as Hydrazide has great significance in organic chemistry. This is the reason why methods of preparation of different types of compounds that are useful in many fields are studied here. It is characterized by containing an active group, which is (-CONHNH2), which consists of two nitrogen atoms attached to four substitutes, and at least one of these substitutes must be an (acyl) or (aryl) group. It is known that these compounds have very important medical, pharmaceutical and industrial functions and applications. Some of them have been employed in the treatment of many common and dangerous diseases, such as respiratory diseases and malaria. In addition, they are highly useful in preparing some important drugs used in the treatment of some chronic diseases related to blood vessels and muscle spasms, showing a wide variety of biological activities, including antibacterial, mitigating, anticonvulsant, antiviral, and antiprotozoal activity. Moreover, they may be used as raw materials in preparing many classes of heterocyclic compounds, including oxadiazole and diazole compounds. Finally, they show significance in industrial applications, particularly in the manufacture of dyes, agricultural pesticides, and growth stimulants for plants and Agricultural crops.

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Piperazine compounds mediate their anthelmintic action by generally paralyzing parasites, allowing the host body to easily remove or expel the invading organism. It is an anthelmintic drug in human as well as in veterinary medicine. This review article represent the various analytical methods which has been reported for estimation of piperazine. The colorimetric ,  spectrophotometric techniques and chromatographic methods like HPLC and RP HPLC, GC, LC-MS, LC-MS/MS and another methods  were reported.