Phytochemical Screening and Nanosynthesis of Linum Usitassimum [Flax Seeds] Used in Hypothyroid Treatment

Introduction

The thyroid is a small  butterfly –shaped gland is positioned in front of neck, just below the voice box (larynx).. This gland plays a completely important role in controlling our frame metabolism i.e The rate at which our frame uses electricity and it does this through producing thyroid hormones like thyroxin or T4 and Triode thyronine or T3 , chemical compounds that travel  through  our blood to every components of our  body. It produces chemical compounds that help the frame to control metabolism .Thyroid hormone is commonly produced in response to every other hormone launched via the pituitary gland¹Generally, thyroid trouble  are grouped in to two most important classes  i.e   Hyperthyroidism  (too much thyroid hormone ),and hypothyroidism,(too little thyroid hormone) right here we discuss approximately hypothyroidism,  the most not unusual signs of hypothyroidism includes  fatigue ,depression , constipation ,weight advantage ,gradual coronary heart rate and so forth. Medicinal plants are the richest bio-resources of medication for classic device of medicine.Flax seeds (Linum usitastassimum L)  is a multipurpose crop and its intake is beneficial for fitness^.2 Flax seeds is a wealthy supply of healthy fats, antioxidant and fibre. The nutrient in flax seeds may additionally assist decrease danger of diabetes, most cancers, heart ailment and thyroid³.Flax seeds is one of the oldest vegetation, having been cultivated due to the fact the beginning of civilization . The Latin name of the flax seed is Linum usitatissimum, which means very beneficial . Flax seed is one of the high content  of omega -three fatty acids ie. Alpha linolinic acid (ALA) ⁴and liganans (phytoestrogens).

The crucial flax seeds developing international locations are Canada ,China, U.S, India and Ethiopia.

Flax seeds has potential health benefits besides the nutrition , due to mainly three reason ; Firstly due to rich source of omega -3-linolenic acid .Secondly being rich in dietary soluble and insoluble fibres. And third ,due to its high contents of liganins, acting as antioxidants, phytoestrogens ⁵. The healthy use of all the three omega- 3 fatty acids have been mostly indicated in several conditions like cardiac diseases, hypertension, ,diabetic condition  osteoporosis, and in many autoimmune conditions and neurological disorders ⁶ . The medicinal importance of a plant is due to the presence of some specific material like alkaloids , glycosides, resins,volatile oil,gum, and tannins etc. Considering all these valuable points  the present study is designed to investigate the presence of various phytochemicals in the flax seeds ⁷. In todays condition AgNO₃ can be prepared by different methods like biologically or chemically.These nanoparticle are of prominent use in many fields like industrial, pharmaceutical, and medicinal field.

In the present work, the silver nanoparticles were prepared from  the flax seeds extract and it is characterized by UV-Vis spectroscopy, Sem analysis with EDX, and FTIR analysis.

Material and method

Collection of seeds

The seeds of flax  were accumulated from local market of Bilaspur (C.G) India.

Extraction  of seeds oil- The seeds were nicely wiped clean and air dried. The dried sample had been then powdered with the assist of house keep grinder. The powdered pattern became stored in an air tight box. Extraction of seeds oil is accomplished by using using sohxlet apparatus the use of petroleum ether as solvent. The extract obtained become gathered and filter via Wattmann No.1 clear out paper to take away alldebris  and unextracted count number. Filtered extract become concentrated the usage of rotatary evaporator at 40 diploma centigrade to acquire focused  extract ⁸.

Phytochemical screening of Flax seeds

Qualitative and quantitative evaluation of flax seeds were finished by way of using general strategies.

Qualitative  analysis of phytochemicals in flax seeds

Chemical test are performed by using standard method of Harborn ,1984.⁹

Quantitative Analysis of flax seeds

Quantitative  check had been accomplished by way of using standard technique of Harborn 1973^.10,11,12

Chromatography Analysis

Preparation for TLC analysis  – Small quantity of seed powder (25 mg) have been dissolved in 1 mL methanol.  Different mobile section at distinctive percentage were taken for the analysis. In case of flax seeds unique cellular solvent gadget have been taken for Preparing take a look at:- According to Harborne¹⁰ by means of the usage of to separation solvent and the lignan was appeared as dark spot on TLC plate  as shown in fig -1.Various  separation solutions like (ethyl acetate: Water) in ratio (1:19) and (Benzene: Methanol) in ratio  (1:9) . The dark spot  has Rf: 0.45 and this  Rf value is equal to Rf value of lignin, according to Harborn [7], when used the liquid – liquid system to separate the lignan from flaxseed. The ethyl acetate : water system has weak polarity and the lignan is soluble in  water and when we evaporate water yellowish powder had get. ¹³

Nanosynthesis

In  present world, nanotechnology a moderately  new field, Nano, a logical word is utilized for deciding the size of the molecule¹⁴ Nanosynthesis, an idea in the field of research and innovation, lately, has likewise been probably going to become dependent on their interest, as different advances.

Nanoparticles are normally a bunch of atoms running between 1-100 nm in  and they display new properties dependent on their size, conveyance & morphology¹⁵ . Numerous materials are combined in nano size for different uses including medical field, scientist in the field of nanotechnology are finding that metals  nanoparticles have a wide range of  surprising advantages. They are normally set up by honorable metals, that is, Ag, Au, pt and pd while silver nanoparticles (AgNPs) are mostly exploited¹⁶ due to its more extensive scope of use in medication, hardware, energy saving, climate, material, beautifiers, biomedical, and so forth.

Preparation of aq.AgNO₃

AgNO₃solution (1 mM) is prepared and saved in dark coloured bottles.

Synthesis of AgNPs

seeds extract of 1 to 5 ml have been taken separately and in this extract 10 mL of 1 mM AgNO₃ solution become delivered with steady stirring and exposed to one of a kind situations like sunlight irradiation, UV irradiation.The colour change of the solution was periodically checked. The colour exchange of the seed extract from  light yellow to dark brown indicates that the AgNPs have been prepared from  seeds extract.This nanoparticle extract are  subjected to centrifugation at 10,000 rpm for 15 min. The pellet prepared were treated with 0.1 mL of toluene H₂O and dried in air.

Characterisation of silver nano particles

U.V spectrophotometer

The prepared AgNPs  were checked by estimating the UV-Vis range of the decrease media in the wake of weakening a little aliquot of the example in distilled water. UV  spectroscopy was done on Systronics UV noticeable spectrophotometer: and checking the spectra between 200-700 nm at the goal of 1 nm¹⁷

SEM and EDAX examination

Scanning electron Microscopic (SEM) investigation was finished utilizing ZEISS EVO 18 special ¹⁸.Edition machine. Additional arrangement was eliminated utilizing smudging paper and afterward films on SEM matrix were permitted to dry by putting it under mercury light for 5 min.

FTIR Analysis

In(FT-IR) Technique Fourier transform infrared (FTIR) spectrophotometer is used to detect functional groups involved in synthesis of nanoparticle at and this study were carried out in the flaxseed hydroalcoholic extract. Fig. (6) shows FT-IR spectra of flaxseed hydroalcoholic extract. The result of FTIR analysis of this work show various bond streaching  at different peak. 2923.89 for OH streaching peaks and 2853.8 for C-H peaks resp.The peak 1740.86  correspond to c=c stretching (non conjugated) . The peak 1460.51 refers to C-H bending. The peak 1374.20 corresponds C-H bending. 1236.87 corresponds to –OH group. 1159.89 correspond to carboxlic acids. 1098.24  streching is correspond to C-O group. The peak 971.85 corresponds to C-group.The peak 722.14 streching  is correspond to >COOH group.¹⁹

Result and Discussion-

 In the present study qualitative analysis of flax seeds shows the presence of alkaloids, phenolic compounds , carbohydrates ,proteins,  amino acid , flavonoids,  saponins . are present in extract of flax seeds while terpenoids is absent in extract of flax seeds. Quantitative analysis of flax seeds shows  alkaloids 0.16 mg , saponins 0.41 mg, phenols 0.23 mg, flavonoids 0.17 mg, carbohydrate o.49 mg.of phytochemicals. Due to presence of these phytochemicals the importance of flax seeds in medicinal field increasing day by day.

Table 1: Qualitatative analysis of flax seeds extract

Alkaloids	Mayer’s test	+
	Wagner’s test	+
	Hager’s test	+
Terpenoids	Salkowski test	–
Phenols	Ferric chloride	+
Steroids	Acetic anhydride+H2SO4	+
Flavonoids	Alkaline reagent test	+
Saponins	Froth test	+
Diterpenes	Copper acetate	–
Coumarine	Sodium hydroxide	+
Carbohydrate	Molish test	+
E modol	25% NH3 solution	–

 

Table 2: Q uatitative analysis of flax seeds extract

Alkaloids	0.16 mg
Saponins	0.41mg
Total phenols	0.23mg
Flavonoids	0.71mg
Carbohydrates	0.49mg

 

Chromatographic Analysis

TLC is a very simple, lowexpensive and  fast method which is mostly  used for a qualitative determination of seed extracts and for determining various stages of lignin seperation. The lignan compound has Rf- 0.45 and this equal to Rf value according to Harborne^{10 .}

Figure 1:  TLC   CHOMATOGRAPHY. Click here to View figure

Nanosynthesis

silver nano particle were prepared by using silver nitrate solution of 1nM through   nanosynthesis and colour change were observed from light yellow to dark brown by continous stirring in magnetic stirrer for 30 min at high temp.

Figure 2: Nanosynthesis of flax seeds. Click here to View figure

After nanosynthesis of seeds extract characterisation of nanoparticle were conducted through different  instrumental methods. Like U.Vspectroscopy  in this analysis absorption band is 348 is observed.

Figure 3: Graph of U.V visible Spectrophotometer. Click here to View figure

SEM and EDX analysis

The results of SEM analysis (Fig. 4) show that the  synthesized silver nanoparticles are of evenly distributed size  and spherical in shape. Thus, the Ag nano-particles  are on evenly distributed throughout the process and also the size and stability of nanoparticles are uniform on the process. The results of EDX helps in determining the elemental composition of the AgNPs (Fig. 5). This analysis helps in confirmation of the presence of the AgNPs in the sample on a varying range between 65-78nm.

Figure 4: Sem analysis of nanoparticle of flax seeds. Click here to View figure

Figure 5: EDX analysis of nanoparticle of flax seeds. Click here to View figure

In present work the result of FTIR analysis show different bond streaching at different peaks like 2923.89 and 2853.85 refers to –OH  stretching and C-H stretching resp.The peak 1740.86  correspond to c=c stretching (non conjugated) . The peak 1460.51 corresponds to C-H bending. The peak 1374.20 corresponds C-H bending. 1236.87 corresponds to –oH group. 1159.89 correspond to carboxlic acids. 1098.24  streching is correspond to C-O group. The peak 971.85 corresponds to C-group.The peak 722.14 streching  is correspond to >COOH group^18.

Figur 6: FTIR analysis of nanoparticle of flax seeds. Click here to View figure

Conclusion

The conducted studies concluded that the presence of various secondary metabolites in flaxseeds extract having medicinal properties and because of their natural properties and enhance qualities by nanosynthesis. It will be beneficial to supplement other medical treatment with natural herbs for underactive thyroids. The conducted studies confirms the presence of functional group that form the part of active compounds of flax seeds which have medicinal properties and can be used as antithyroid agents. Flax seeds extract consist of oil in abundance. The active compound in flax seeds is called Omega -3- fatty acids i.e. Alpha linolinic acid has the power to influence thyroid function and improve the condition of hypothyroidism. It has been reported that the consumption of Omega -3- dietery supplements leads to significant reduction of non steroidal anti inflammatory drugs.¹⁹ Studies have shown that the high intake of dietery fibers is beneficial of obesity in both men and women.²⁰ An advantage of this herb is decrease the level of harmful cholesterol which is due to high weight gain and which is one of the feature of hypothyroidism.

Acknowledgement

The authors would like to acknowledge to Dr. C.V.Raman University for their laboratories facilities. And the authors extend their indebted thanks to metallurgy department of NIT Raipur and Pt. Ravi Shanker University Raipur for instrumental analysis.

Conflict of Interest

There is no conflict of interest among the authors regarding the publication of this manuscript.

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