A NANOFORMULATION FOR IMPROVED BANANA PRODUCTIVITY AND METHOD OF SYNTHESIS THEREOF

Abstract

A synergistic nanoformulation for improved banana productivity and method of synthesis thereof is disclosed. Said synergistic nanoformulation broadly comprises: a spray dried mixture of: copper sulphate, ferrous sulphate, manganese sulphate, zinc sulphate, boric acid, and citric acid, along with a surfactant that comprises 2-[2-(4-nonylphenoxy) ethoxy] ethanol, 1-butanol, and tall oil fatty acid. The disclosed nanoformulation offers at least the following synergistic advantages and effects: requires less concentration (of about 1%); improved efficacy; and/or improved solubility and stability.

Specification

Description:TITLE OF THE INVENTION: A NANOFORMULATION FOR IMPROVED BANANA PRODUCTIVITY AND METHOD OF SYNTHESIS THEREOF
FIELD OF THE INVENTION
The present disclosure is generally related to banana productivity. Particularly, the present disclosure is related to formulations for improving banana productivity. More particularly, the present disclosure is related to: a nanoformulation for improved banana productivity and method of synthesis thereof.
BACKGROUND OF THE INVENTION
Commercial fertilisers for improving banana productivity are already known in the art, but suffer from at least following drawbacks such as: requirement of a high concentration (about 2%) leading to environmental pollution; low efficiency; and/or poor solubility and stability.
There is, therefore, a need in the art, for: a nanoformulation for improved banana productivity and method of synthesis thereof, which overcomes the aforementioned drawbacks and shortcomings.
SUMMARY OF THE INVENTION
A synergistic nanoformulation, for improved banana productivity, and method of synthesis thereof is disclosed. Said synergistic nanoformulation broadly comprises: a spray dried mixture of: copper sulphate, ferrous sulphate, manganese sulphate, zinc sulphate, boric acid, and citric acid, along with a surfactant.
Said copper sulphate is in a ratio of about 1:2.42 with said ferrous sulphate; said copper sulphate is in a ratio of about 1:1.33 with said manganese sulphate and said boric acid; said copper sulphate is in a ratio of about 1:2.17 with said zinc sulphate; said copper sulphate is in a ratio of about 1:0.083 with said citric acid; and concentration of said surfactant is about 3% v/v.
The disclosed nanoformulation offers at least the following synergistic advantages and effects: requires less concentration (of about 1%); improved efficacy; and/or improved solubility and stability.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1A illustrates tissue cultured banana plantlets, in accordance with an embodiment of the present disclosure;
Figure 1B illustrates treated plantlets, in accordance with an embodiment of the present disclosure;
Figure 1C illustrates uprooting of treated plants after 90 days, in accordance with an embodiment of the present disclosure;
Figure 1D illustrates segmenting uprooted plants into leaves, pseudo stems, corms, and roots for dry and wet matter estimation, in accordance with an embodiment of the present disclosure;
Figure 2A and Figure 2B illustrate evaluation of synthesised nanoformulations for dry matter production at different concentrations, in accordance with an embodiment of the present disclosure;
Figure 3 illustrates results of dry matter production analyses (***p<0.0001), in accordance with an embodiment of the present disclosure; and
Figure 4A and Figure 4B illustrate results of micro and macronutrients uptake analyses, respectively, in accordance with an embodiment of the present disclosure (***p<0.0001).
DETAILED DESCRIPTION OF THE INVENTION
Throughout this specification, the use of the words "comprise" and "include", and variations, such as "comprises", "comprising", "includes", and "including", may imply the inclusion of an element (or elements) not specifically recited. Further, the disclosed embodiments may be embodied, in various other forms, as well.
Throughout this specification, the use of the word "synthesised", and its variations, is to be construed as: "produced; manufactured; and/or the like".
Throughout this specification, the use of the phrase "treated plantlets" is to be construed as: "tissue cultured banana plantlets treated with synthesised nanoformulations (NBS9, NBS2, NBS4, and NBS5)".
Throughout this specification, the use of the acronym "NBS" and the word/acronym "Normal BS" are to be construed as: "nano micronutrient suspension for banana" and "micronutrient mixture for banana", respectively. For example, the "micronutrient mixture for banana" is a conventional mixture of micronutrients that is applied on banana crops.
Throughout this specification, the use of the designation "NBS9" is to be construed as: "nano micronutrient suspension for banana without any surfactant".
Throughout this specification, the use of the designation "NBS2" is to be construed as: "nano micronutrient suspension for banana with about 3% surfactant 1".
Throughout this specification, the use of the designation "NBS4" is to be construed as: "nano micronutrient suspension for banana with about 3% surfactant 2".
Throughout this specification, the use of the designation "NBS5" is to be construed as: "nano micronutrient suspension for banana with about 1.5% of surfactant 1 and about 1.5% of surfactant 2".
Throughout this specification, the use of the designation "surfactant 1" is to be construed as a surfactant that comprises: "2-[2-(4-nonylphenoxy) ethoxy] ethanol, 1-butanol and tall oil fatty acid". A person skilled in the art will appreciate the fact that such surfactants are commercially available (for example, AMWAY, India).
Throughout this specification, the use of the designation "surfactant 2" is to be construed as a surfactant that comprises: "Silicone polyether Trisiloxane". A person skilled in the art will appreciate the fact that such surfactants are commercially available (for example, Resil Chemical Private Limited, India).
Throughout this specification, the use of the phrases "reference control" and "Normal BS" is to be construed as: "foliar treatment/spray with conventional formulation".
Throughout this specification, the use of the phrase "absolute control" is to be construed as: "foliar treatment/spray without conventional formulation".
Throughout this specification, the use of the phrase "conventional formulation" is to be construed as: "commercially available fertiliser".
Throughout this specification, the disclosure of a range is to be construed as being inclusive of: the lower limit of the range; and the upper limit of the range.
Throughout this specification, the phrases "nano micronutrient suspension for banana" and "nanoformulation" are used interchangeably.
Throughout this specification, the words "the" and "said" are used interchangeably.
Also, it is to be noted that embodiments may be described as a method. Although the operations, in a method, are described as a sequential process, many of the operations may be performed in parallel, concurrently, or simultaneously. In addition, the order of the operations may be re-arranged. A method may be terminated, when its operations are completed, but may also have additional steps.
A synergistic nanoformulation (also referred to as "synthesised nanoformulation" and/or "nanoformulation",), for improved productivity in banana, is disclosed. The disclosed nanoformulation was synthesised and tested, as follows:
The active ingredients were a spray dried mixture of: copper sulphate; ferrous sulphate; manganese sulphate; zinc sulphate; boric acid; and citric acid, along with a surfactant 1 and/or a surfactant 2. The spray drying was performed at an inlet temperature of about 150 degrees Centigrade, an outlet temperature of about 70 degrees Centigrade; a feed rate of about 15 mL/hour, and an aspiration rate of about 1,200 rpm.
The surfactant 1 solution was prepared by mixing about 3% of surfactant 1 (for example, 0.6 mL) in about 20 mL of distilled water. The nanoformulation was synthesised by mixing the spray dried mixture of said copper sulphate (about 1.2 g), said ferrous sulphate (about 2.9 g), said manganese sulphate (about 1.6 g), said zinc sulphate (about 2.6 g), said boric acid (about 1.6 g), and said citric acid (about 0.1 g) with about 20 mL of a pre-prepared aqueous surfactant 1 solution to obtain a formulation.
Said copper sulphate is in a ratio of: about 1:2.42 with said ferrous sulphate; about 1:1.33 with said manganese sulphate and said boric acid; about 1:2.17 with said zinc sulphate; and about 1:0.083 with said citric acid.
The obtained formulation was stirred at a speed of about 200 rpm using magnetic stirrer for about 5 to about 10 minutes, followed by high-speed homogenisation at a speed ranging between 8,000 rpm to 10,000 rpm for about 30 minutes to obtain the nanoformulation.
Physicochemical Characterisations
Sedimentation Behaviour, Redispersion Time Analyses, and Clouding Analyses
A volume of about 20 mL of the below nanoformulations were evaluated for homogeneity at time t=0 using a graduated measuring cylinder. Sediment height, volume, and sedimentation time were recorded, followed by vertical inversion and about 180° manual rotation of the tube, and the results are given in below table.
Designation Sediment Time (min) Sediment Volume (mL) Redispersion Time (Sec)
NBS2 6.666 ± 1.527 2.833 ± 0.76 4.16 ± 0.28
NBS4 9.666 ± 3.511 4.033 ± 0.152 4.5 ± 0.5
NBS5 - - 8.33 ± 0.57
NBS9 120 ± 4 2.266 ± 0.472 18.66 ± 1.15

Further, the above nanoformulations were vigorously shaken for about 2 to about 3 minutes in graduated vials and then allowed to settle at room temperature. Decrease in turbidity in the upper phase was measured to calculate cloud volume, while the time for cloud formation was visually recorded and the results are given in below table.
Designation Cloud Time (min) Cloud Volume (mL)
NBS2 - -
NBS4 - -
NBS5 4.33 ± 0.57 3.66 ± 0.57
NBS9 - -
Viscosity, pH, Average Particle Size, PDI and Zeta Potential Analyses
Viscosity of the below nanoformulations were measured with a viscometer using spindle number 64 at about 200 RPM.
Zeta potential of the nanoformulations were evaluated at about 25 ˚C with water as a dispersant, and estimated using Smoluchowski equation. Average particle size was determined by Stokes-Einstein equation.
The results of physicochemical characterisations of the synthesised nanoformulations are shown in the below table.
Designation Viscosity
(Ns/m²) pH of Nanosuspension Concentrate pH of Diluted Foliar Spray Average Particle Size (nm) PDI Zeta Potential (mV)
NBS2 5.73 ± 0.3 2.07 - 2.96 5.65 - 5.67 336.4 0.529 -16.6
NBS4 4.83 ± 1.7 1.81 - 2.00 5.60 - 5.65 402.2 0.573 -19.6
NBS5 5.22 ± 2.1 1.85 - 1.94 5.52 - 5.66 321.7 0.436 -14.4
NBS9 4.47 ± 0.4 1.95 - 2.02 5.57 - 5.61 1149 0.871 -3.24
Real-Time Stability Analyses
Real-time stability of the below nanoformulations was determined for duration of about 6 months at temperature of about 25°C ± 2°C and relative humidity of about 60% RH ± 5% RH, as shown in the table below.
Designation Sedimentation Time
(min) Sedimentation Volume
(mL) Clouding
Time
(min) Clouding Volume
(mL) Redispersion Time
(Sec) Viscosity
(cP) pH of Nanosuspension Concentrate pH of Diluted Foliar Spray
NBS2 6.77 ± 0.53 2.85 ± 0.04 - - 4.18 ± 0.166 5.68 ± 0.14 2.75 - 2.86 5.31 - 5.42
NBS4 10.83 ± 0.72 4.08 ± 0.14 - - 4.53 ± 0.31 4.67 ± 0.32 1.65 - 1.86 5.38 - 5.45
NBS5 - - 4.69 ± 0.40 3.89 ± 0.20 8.6 ± 0.58 5.29 ± 0.31 1.73 - 1.95 5.43 - 5.51
NBS9 12.53 ± 1.40 2.30 ± 0.04 - - 30.5 ± 1.03 4.32 ± 0.14 1.85 - 2.16 5.32 - 5.44
Accelerated Stability Analyses
Accelerated stability of the below nanoformulations were determined under stressful conditions of high temperature at about 40°C ± 2°C, relative humidity or light at about 75% RH ± 5% RH, where the rate of decomposition would be high, as per the ICH guidelines, as shown below.
Designation Sedimentation Time
(min) Sedimentation Volume
(Cm) Clouding Time
(min) Clouding Volume
(Cm) Redispersion Time
(Sec) Viscosity
(cP) pH of Nanosuspension Concentrate pH of Diluted Foliar Spray
NBS2 6.61 ± 0.31 3.08 ± 0.29 - - 4.92 ± 0.67 5.55 ± 0.50 2.16 - 2.85 5.57 - 5.94
NBS4 10.34 ± 0.52 4.85 ± 0.71 - - 4.63 ± 0.81 5.23 ± 0.35 1.65 - 2.56 5.30 - 5.78
NBS5 - - 5.32 ± 0.78 4.31 ± 0.56 8.43 ± 0.43 5.30 ± 0.37 1.76 - 1.83 5.68 - 5.73
NBS9 12.1 ± 0.60 3.2 ± 0.73 - - 30.12 ± 0.71 4.76 ± 0.71 1.92 - 2.05 5.53 - 5.72

 
Total Dry Matter Production Analyses
As illustrated, in Figure 1A, tissue cultured banana plantlets were selected. The synthesised nanoformulations - NBS2, NBS4, NBS5 and NBS9 were diluted with water to obtain different concentrations of about 0.25%, about 0.5%, about 1%, and about 2%. About four replications of a reference control and absolute control were also maintained.
The selected plantlets were then sprayed with about 4 mL to about 8 mL of diluted NBS2, NBS4, NBS5 and NBS9, at weekly intervals, as illustrated, in Figure 1B. Foliar spray application was performed for about 12 times during the total study period of about 90 days.
All the plants were uprooted and segmented into leaves, pseudo stems, corms, and roots at the end of about 90 days, as illustrated, in Figure 1C and Figure 1D.
As illustrated, in Figure 2A, Figure 2B, and the table given below, the total Dry Matter Production (DMP in g/plant) was calculated.
Spray
Solution
Concentrations Treatments Mean
NBS-9 NBS-2 NBS-4 NBS-5 Normal BS
0.25% 12.4 13.3 7.0 12.0 12.1 12.4
0.50% 13.7 11.5 6.7 12.3 12.2 11.3
1% 4.9 14.5 3.6 9.4 10.7 8.6
2% 1.7 9.1 3.6 10.5 12.0 7.4
Mean 8.6 11.7 6.2 10.8 11.4
It was established from the above table, and as illustrated, in Figure 3, that the synthesised nanoformulation NBS2 resulted in an enhanced dry matter content, at a low concentration of about 1%.
Nutrient Uptake Analyses
Uptake of various macro and micronutrients such as nitrogen, phosphorus, potassium, calcium, magnesium, iron, copper, manganese, zinc, and boron were quantified.
As illustrated, in Figure 4A and Figure 4B, said NBS2 synthesised nanoformulation resulted in statistically increased uptake of iron, copper, manganese, zinc, boron, potassium. The uptake of nitrogen and magnesium from NBS2 was equivalent to the normal banana suspension.
The disclosed nanoformulation offers at least the following synergistic advantages and effects: requires less concentration (of about 1%); improved efficacy; and/or improved solubility and stability.
It will be apparent to a person skilled in the art that the above description is for illustrative purposes only and should not be considered as limiting. Various modifications, additions, alterations, and improvements, without deviating from the spirit and the scope of the disclosure, may be made, by a person skilled in the art. Such modifications, additions, alterations, and improvements, should be construed as being within the scope of this disclosure.
Acknowledgements
The Inventors and the Applicants thank the Department of Biotechnology, Government of India, for funding support.
, Claims:1. A synergistic nanoformulation, for improved banana productivity, comprising: a spray dried mixture of: copper sulphate; ferrous sulphate; zinc sulphate; boric acid; and citric acid; and a surfactant that comprises 2-[2-(4-nonylphenoxy) ethoxy] ethanol, 1-butanol, and tall oil fatty acid, with:
said copper sulphate being in a ratio of 1:2.42 with said ferrous sulphate;
said copper sulphate being in a ratio of 1:1.33 with said manganese sulphate and said boric acid;
said copper sulphate being in a ratio of 1:2.17 with said zinc sulphate;
said copper sulphate being in a ratio of 1:0.083 with said citric acid; and
concentration of said surfactant being 3% v/v.
2. The synergistic nanoformulation, for improved banana productivity, as claimed, in claim 1, wherein:
weight of said copper sulphate is 1.2 g;
weight of said ferrous sulphate is 2.9 g;
weight of said manganese sulphate and said boric acid is 1.6 g;
weight of said zinc sulphate is 2.6 g;
weight of said citric acid is 0.1 g; and
volume of said surfactant is 0.6 mL in 20 mL of distilled water.

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