Performance SWOT of Micronized Voriconazole to Augment Health Security and its Bioassay

Nowadays fungal diseases certainly affect people with undermined immune system. Fungal infections are most likely impinge on skin, lungs, nails etc and it leads to systemic infection. Voriconazole (VCZ) is an antifungal medication of the triazole class used to treat infections caused by a fungus. VCZ is used to treat patients suffering with invasive fungal infections. The focus of this paper was to study the performance swot of antifungal activity of untreated VCZ and micronized VCZ. Size reduction of drug treated as an imperative operation to make handling of the drug easier, increase surface area per unit volume, and separate entrapped components. The size reduction process may involve accelerating particles so either particle-to-particle impact. Biological evaluation study was conducted by using Aspergillus niger and Candida albicans as test organisms’. Zone of inhibition was measured to assess antifungal activity by adopting agar diffusion process. The antifungal activity was conducted to the untreated VCZ material having particle size, about 207 microns and micronized VCZ having particle size 12 microns. The determination of size distribution of VCZ particles was examined through Dynamic light scattering (DLS) and Scanning Electron Microscopy (SEM) for treated and untreated VCZ.The structural identification study was conducted by Ultraviolet spectroscopy (UV) and Fourier transform Infrared spectroscopy (FT-IR). The anti fungal activity effectiveness of micronized VCZ illustrates increased efficiency when compared to untreated VCZ material.


INTROdUCTION
Vo r i c o n a z o l e ( C 16 H 14 F 3 N 5 O ) ( V C Z ) is a antifungal drug. VCZ has an incomparable bioavailability, stumpy toxicity, and activity against many pathogenic species. VCZ belongs to triazole class and used orally for systemic fungal infections. VCZ is particularly active against Aspergillus species and Candida species. The VCZ drug exhibits wide spectrum fungicidal activity against molds and fungistatic activity. As like all azole agents, VCZ is inhibition of cytochrome P450 (CYP 450)-dependent 14α-lanosterol demethylation, which is a vital step in cell membrane ergosterol synthesis by fungi 1 . Symptoms of overdose include abnormal vision, nausea, headache, vomiting, dizziness and dry mouth. was necessary for the treatment of patients with leukemia 9 . Roya Vahedi-Shahandashti describes novel antifungals, with a focus on the Aspergillus species 10 . Mahmoud has studied the increased use and availability of different classes of antifungal agents 11 . The studies reveal the oral bioavailability reaches approximately 95% and safer to use 12 . Researchers have found that VCZ, used to treat fungal infections after lung transplantation, increases the risk of developing skin cancer by 73% 13 . Different studies exemplify VCZ bioassay using Aspergillus niger and Candida genus. The main focus of the present study intended to appraise the action of micronized VCZ to augment health safety.

Materials
Voriconazole working standard was provided for the research work from Hetero Labs Ltd, Visakhapatnam, and Andhra Pradesh, India.
The fungi strains were obtained from GVK Bio. Sci, Hyderabad, Telangana state, India.
All other chemicals used in the present study were of Analytical reagent grade.

Instrumentation
Voriconazole (VCZ) is a second-generation triazole that has an enhanced antifungal spectrum, compared with older triazoles like fluconazole and itraconazole [2][3][4] . Reduction in the therapeutic time and/ or increase the quality of life of patients and tolerance was possible when new antifungal strategies developed [5][6] . VCZ was acceptably tolerated, despite the diverse nature of the patients studied and their severe illnesses 7

Preparation of plain and micronized VCZ
The untreated VCZ material was taken into dry micronizer equipment (Microtech-M50 shown in Fig. 2). With the help of moisture free Nitrogen.the VCZ sample was dispensed into micronizer hopper with a feed rate of 4 Kg air pressure and 7 Kg micronization pressure. Similar method was repeated for two cycles to get maximum micronized material. The impurities present in the final sample are found less than 0.1%. The loss on drying was 0.15% w/w. The particle size was drastically reduced during micronization process. The particle size information was presented in Table 1.

RESULT ANd dISCUSSION
For the treatment of fungal infections antifungal drug performance was crucial. Benefits of particle size reduction of drug materials were increased surface area, downy surface quality and elevated-quality product utility. The surface modification of VCZ particles showed significant augmentation of antifungal activity.

Laser diffraction measurement of particle size analysis
Laser diffraction has become the most widely used particle characterization technique in the pharmaceutical industry. The DLS instrument applied by Particle Analytical has a flow-through cell. The VCZ is a free-flowing powder and nonhygroscopic, the dry mode sampling technique is used to determine the particle size measurement. The measurement was made using 1.0 bar air pressure with a 50% feed rate of specimen sample. The micronized sample resulted to 12 microns from 200 microns, after reduction of particle size using micronization process.

SEM Analysis
To know the size of micronized VCZ and surface topology at diverse magnifications scanning electron microscope was selected. A small pinch of micronized VCZ was taken on the sampler slide. By using sputter coater the sample was subjected to sputtering. This was obtained when the area of specimen intermingled with the electron beam produces signals. Generally in SEM an electron gun produces electron and later accelerated through the condenser. SEM images were shown in Fig. 4 demonstrates the shape of micronized VCZ at a range of magnifications in which micronized VCZ particles appeared as clusters. From SEM images it was found that the size is around 2 microns and shapeless, since the powder is fine.

FT-IR Study
For categorization of the functional groups a diminutive amount of untreated and micronized VCZ in Potassium bromide (KBr) pellets in transmittance mode was processed. About 2.5 mg of VCZ sample was mixed with 250 mg of KBr and grounded to make an apparent disc. Scanning process was done and the respective spectra were recorded. From FT-IR spectra the chemical structure of micronized VCZ material was not changed. The untreated and  The informative spectral ranges were examined, the characteristic peaks corresponding to structure were identified.The bands of OH stretching at 3193 cm −1 , C-N stretching at 1507 cm −1 , and C-F stretching at 1585 cm −1 , respectively were identified as a compound functional group. Observation: The structure of the micronized material is similar to plain material and does not have any impact on structure due to micritisation.

Identification by UV Spectroscopy
The identification of Voriconazole is done using 10 ppm concentration of both Plain and Micronised Voriconazole was prepared and analysed using UV (Shimadzu model) spectrophotometer. The absorption maxima are using beers lamberts principle was observed at wavelength of 255 nanometers. The plain and micronized material showed similar absorptions bands at similar wavelengths. From Fig. 7 and 8 it was concluded that the material identity has no disturbance even after micronisation.

Bioassay of VCZ (Agar diffusion technique)
Fungi like Aspergillus niger and Candida albicans are the two mainly widespread fungal pathogens causing ruthless invasive diseases. The pathogenic strains stored at 40 0 C under test were sustained on agar slants (nutrient). 25% glycerol was used for long period storage of slants preservation. To begin with the medium and glass petri-dishes were autoclaved for 25 min by the application of 15 Pa pressure. The methodology incorporates the injection of bacterial cells on nutrient agar Petri dishes. The untreated and micronized VCZ samples are laid over these dishes. The dishes are incubated for 18-24 h at 37°C and subsequently the growth of bacteria is determined. Precisely measured the diameters or areas of the circular inhibition zones and addressed in Table 2. The images are addressed in Fig. 9.The effectiveness of untreated (or plain) and micronized VCZ material against fungi was given in Table 2.

CONCLUSION
Reduction in particle size of VCZ was shown better results as compared to untreated VCZ material. The efficiency of micronized Voriconazole antifungal activity against Candida albicans and Aspergillus niger demonstrates increased efficiency, when compared to untreated VCZ material due to micronization process that resulted in very fine particles.