Welcome to Biological Agri Solutions Association of India
To define guidelines for sharing of data generated by scientists in India using modern biotechnological tools and methods.
Although, this document provides broad guidelines for biological data in general, it specifically pertains to modern high-throughput, high-volume data, for example, data generated by nucleic acid sequencing and microarrays, biomolecular structures and flow cytometry.
The Government of India invests a large amount of money to generate data in various sectors, including the biotechnology sector. These data are generated in the context of furthering knowledge, gaining deeper insights into biological and other processes and for translation. Ultimately, all data are to be used for the benefit of humankind; that is the major reason for investment of public funds to generate data. Sharing of data maximizes the collective utility of data.
However, there are many issues that must be taken into account in the context of data- sharing, the most important of which is that data-sharing must be done in a responsible manner. Data may induce vulnerability to individuals and to populations. The rights to privacy and confidentiality of individuals and populations must be protected as emphasized in the U.N. Declaration of Human Rights, and no harm must be done to them as a result of data-sharing.
This document provides a framework and principles for sharing of data while protecting the rights of individuals and populations and without causing any harm to them.
Data generated from public funds are for public good. Data are, therefore, a resource for human development. Unless the data are shared publicly and within a reasonable period of time after data-generation, the utility of the data will be constrained. Resultantly, accrual of benefit of public investment for data generation will be compromised. The necessity of data-sharing is, therefore, to accrue maximal benefit from public investment in generation of data.
Since data are a public resource, there are primarily three stakeholders of this resource – funders who help generate this resource, producers and users of this resource. All three stakeholders must assume responsibility on how the data may be shared. Even though data are a public resource, sharing of data have ethical implications. Data pertain to
individuals and contain private information. When such data are shared, there may be breach of privacy. Therefore, sharing of data must be done in a responsible manner. Modalities in which data are shared must protect privacy, confidentiality, security and should be non-discriminatory and fair. These issues have been emphasized in the National Ethical Guidelines for Biomedical and Health Research involving Human Participants, 2017, established by the Indian Council of Medical Research (ICMR); it is expected that these guidelines are followed in research involving humans.
Responsible data-sharing implies that certain principles are to be followed. These include,
The quality of the data must be of a high standard, unbiased and verifiable. The data submitter is responsible for ensuring high quality and authenticity of submitted data. The storage must be done in a manner that protects privacy and confidentiality, and promotes ease of access, search and long-term maintenance. Appropriate security features must be embedded in the storage and access framework to avoid breach of data-trust. Features to enable tracing of chain of data access may be built-in. The storage and security policy must also address duration of data storage and accessibility. Mechanisms for obtaining feedback from resource users must be put in place in order to improve data quality, data access, data integrity and interoperability.
Data-sharing policy must be transparent and must state in a publicly-accessible manner the policy of data transfer within and across national boundaries, with public and private organizations, for knowledge and commercial use, etc.
2.3 Harmonization with international policies, ensuring that national policies supersede
Many international consultations have been held to establish and evolve norms, rules and regulations for data sharing. These include the Bermuda Agreement, 1996; Fort Lauderdale Agreement, 2003; Nagoya Protocol, 2011, guidelines of the Global Alliance for Genomics and Health (GA4GH), 2013 and the European Union’s General Data Protection Regulation, 2016. This current data-sharing policy respects and upholds the principles and tenets of the international discourses and agreements. These discourses have emphasized, in the main, the principle that data should be rapidly released after generation. The present data- sharing policy also strongly supports this principle. An Indian National Data-Sharing and Accessibility Policy was promulgated in 2012. While the national data-sharing policies are generally harmonized with the international policies, it is emphasized that in a circumstance in which there is a misalignment of national and international policies, national policies shall supersede.
2.4 Data-sharing: Open access, Managed access, No access Data resource sharing can be of three types.
3.2.2 RNA sequence transcriptomic data – The nature of the data are similar to those generated by a massively-parallel DNA sequencer, since usually cDNA synthesis is performed before sequencing. However, recent technological developments allow single-molecule direct RNA sequencing without cDNA synthesis.
3.2.3 Genotype data – Modern methods use high-density microarrays to genotype individuals at a large number of loci spread across the entire genome. Genotyping by sequencing (GBS) is being increasingly used for genome wide association studies especially in plants. However, for various specific purposes, small-scale genotyping using PCR-RFLP and other similar technologies continue to be used.
3.2.4 Epigenomic data – These data are also primarily generated using a BeadChip (that is similar to a DNA microarray). However, epigenomic data may also be generated using sequencing methods after bisulfite conversion.
3.2.5 Microbiome data – These data are also nucleic acid sequence data and currently are of two types (a)Amplicon sequencing data from which specific groups of microrganisms present in any sample (e.g., human stool, soil, sediment, etc.) can be identified, or (b) Shotgun metagenomic sequence data that allows comprehensive assessment of all microbial organisms present in a sample.
3.2.5 Protein Structure data – Atomic coordinates and other information that describes a protein and other important biological macromolecules comprise such data. These data provide 3D shapes of proteins, nucleic acids, and complex assemblies that help understand various aspects of protein synthesis under different conditions.
3.2.6 Mass Spectrometry data – Mass spectrometry (MS) is a key analytical technology in current proteomics and mass spectrometers are widely used to generate data that allow protein identification, annotation of secondary modifications, and determination of the absolute or relative abundance of individual proteins.
3.2.7 Flow Cytometry data – Flow cytometry is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. Flow cytometry data pertain to counts and multi-parameter profiles of different types of cells in a heterogeneous fluid mixture.
3.2.8 Imaging data – Images of individual cells, organs or body parts, for example, chest X-rays or images of human eyes or mouth cavity.
3.2.9 Metabolome data- Metabolomics is increasingly used in conjunction with microbiome data to better understand host-microbiome interaction. Small molecule metabolite patterns are generated using either LC MS, GC MS or CE MS.
Examples of common high-throughput data-types are provided in Appendix-1.
4. Framework for Data Sharing and Access
(a) Data generated from publicly-funded projects should be shared openly for public good, with few restrictions and in a timely manner, safeguarding the ethical issues that may arise out of shared data.
(b) High standards and best practices should be used in generation, management and access to data. Data that are valuable in the long-term should be stored in a manner that these remain accessible for a long time.
(c) Shared data will always be de-identified
(d) Under specific circumstances, even data generated using public funds may not be provided open access, and may be provided under a managed/controlled access protocol.
(e) To enhance use of data, metadata must also be released in a timely manner.
(f) Access to data that are of “sensitive” nature may be barred, even if generated using public funds.
(g) The conduct of research must not be jeopardized by release of data. The research organization must ensure that due consideration is given to protect the interest of the data generator.
(h) Data generator may require privileged use of the data. Therefore, there may be a period of moratorium before the data generator releases the data in the public domain. The period of moratorium may vary with the nature of the data; public resource data need to be released without any significant time lag.
5. Data Release Strategy and Timing of Data Release
5.1 Data Release and Timing
In current research and other scientific activities, large volumes of data are generated. These data comprise raw data that are produced by the various equipment that are used, e.g. DNA sequencer, Flow cytometer, etc. The raw data are then processed and analysed by researchers to draw scientific inferences. When public funds are used to generate data, these data must be made accessible to others in a form that is valid and user-friendly. It is recognized that raw data can be shared almost immediately after it is generated, but data- processing may take time and hence processed data may not become immediately amenable to sharing. Further, often there is no unique method of data-processing; methodological development may also be a part of data-processing.
It is recommended that – when funds provided by any agency of the Government of India to generate data, either wholly or partially – data, after appropriate clean-up and curation, be shared in accord with the following guidelines:
5.1.1 Raw (Level-1) data must be shared, by placement on a database identified and approved by the funding agency of the Government of India, within one year of generation of the data. If no such database is identified by the agency, then raw data must be made available to anyone working in any Indian institution, public or private, requesting for these data. The sharing of raw data must also include the experimental conditions and specifications of the equipment used to generate these data (experimental metadata), where relevant.
Sometimes an agency of the Government of India funds activities that are solely devoted to data generation, usually for the purpose of generating a “reference” data set. When data from such a project are generated, these data must be released within six months of data-generation.
5.1.2 Processed (Level-2) data based on data generated wholly or partially with funding from Government of India must also be shared. In recognition of the facts that (a) processing of data takes time, and (b) the research group that was funded to generate data and draw inferences from the data must be accorded the first right to publish the findings, it is recommended that processed data may be shared with others within two years of data-generation.
5.2 Release of Metadata
Use of certain types of data even if made publicly available may be of limited use unless some associated metadata are also made available. Such metadata include gender, ethnic background, phenotype, etc. Metadata should be released concurrently with other types of data (e.g., DNA sequence data) in order that the value of the released data is not diminished.
5.3 Data Deposition
In addition to releasing data, it is the responsibility of the data-generator to deposit data in an appropriate database in a National Biological Data Centre, as identified by the Department of Biotechnology. Until a National Biological Data Centre is established, raw and processed data must be stored on an institutional data storage facility. The data should be made available to anyone working in any Indian institution, public or private, who may request access to these data. Along with sharing of processed data, details of relevant methods used for processing raw data must also be shared.
5.4 Exemptions to Data Release (“Sensitive” data)
Release of data that compromises or impacts on national security shall be exempted. There may also be other circumstances when data-release exemption may be granted. There are some tribal populations in India that are numerically small. Whole genome sequence data released on individuals from such populations, with or without metadata, can result in individual identification. Therefore, if an exemption to release of such data is requested, the request may be considered and granted.
5.5 Withdrawal of Data
An individual donor whose data have been placed on a publicly accessible database may request for withdrawal of data, even if the donor had provided consent initially. Such requests may be considered and granted provided that the data are identifiable in the database.
6. Data User Agreement
Most data stripped of all personal identifiers and data that are not subjected to any intellectual property or patent restrictions should be made accessible openly (open access data), especially if the data are generated using public funds. Sometimes, data generated in even in publicly-funded projects may not be allowed open access for a variety of reasons, prominent among them being that the data provider may need to be recontacted or may belong to a vulnerable subgroup or intellectual property issues may be under consideration. Such data should still be made available to others under managed access, that is, data accessibility should be provided only if the data-requester provides sufficient justification to request access the data and the purpose of data-use is valuable and ethically appropriate.
An individual requesting data that are accessible may do so using a Data Request Form (Appendix-2).
A Data Usage Agreement Form must be signed by the data recipient using the form provided in Appendix-3.
6.1 Open Access Data
6.1.1 Acknowledging the data provider: If the data-provider is identified in the database, then it is expected that the data user will adequately acknowledge the data-provider in publications and such documents in which results generated from the data are announced.
6.1.2 Who shall hold intellectual property arising from the shared data?: The onus of arriving at this decision is on national regulatory authorities and to a large extent depends on prior intellectual property rights granted by regulatory authorities to others, notably the data-generator.
6.1.3 Will the data be shared with others?: Open access data are public and are shared with anyone interested in accessing the data.
6.1.4 Will efforts be made for re-identification of individuals who may have provided data for inclusion in the database?: Re-identification is prohibited for open access data.
6.1.5 Legal Issues
22.214.171.124 Who will be held liable for data misuse?: If legal provisions are invoked for data misuse, such decision will be made by the court of law.
Otherwise, national funding agencies or peer groups (e.g., national science academies) may consider the nature of misuse and provide suitable reprimands.
126.96.36.199 If there is a dispute, how to resolve?: If the dispute is escalated to a court of law, then national legal modalities will apply for resolution. Otherwise, national funding agencies or peer groups (e.g., national science academies) may provide a platform for negotiation and resolution.
188.8.131.52 Duration of data access: There is no upper limit to the duration of access to open-access data. However, technologies (e.g., data storage space) may be the determining factors to limit the duration of access.
6.2 Managed Access Data
6.2.1 Purpose of access: Description, Ethics approval: As mentioned above, managed data usually have ethical, intellectual property and similar issues attached to the data. Therefore, unless the need for data-access outweighs the burden that may arise from these issues, access to such data may not be provided. Modalities of management of access to such data are usually established by the institution responsible for data-generation. Therefore, the data-requester must apply for data- access by providing a detailed description to the data-management group for reasons to request access to the data, the possible uses to be made of the data and the ethical precautions to be followed during and after data access.
6.2.2 Competence of researchers requesting data access: The data-management group shall assess the competence of the researchers to responsibly use the data for the purposes described by the data-requester before access to the data is provided.
6.2.3 Authority designated to sign on behalf of the data user: Unless otherwise stated, normally the head of the institution to which the data-user belongs or a designated nominee shall sign applications and other documents pertaining to data access and use on behalf of the data-user.
6.2.4 List of users authorized to access the data: Access to managed-data shall normally be given to a one or a small number of users of an institution. The list of persons who plan to access and use the data shall be provided on the application for data-access. The data-management group shall examine the list of possible users and their levels of competence before providing approval to data-access.
6.2.5 Duration of data access: The duration may be variable depending on intent of use of the data. The duration for which access is requested must be specified in the application and the data-management group may examine the appropriateness of the duration for which data-access is requested before approval.
6.2.6 Whether renewal of data access may be sought?: Yes. A fresh application must be submitted to the data-management group, in which the past use of the data and the future intended use shall be clearly described and justified.
6.2.6 How will confidentiality and security of shared data be ensured?: The application for data-access must clearly describe the plan to uphold the confidentiality of the data and the security of the data to prevent access by unauthorized users.
For open-access data, there may be a national committee established by a consortium of national funding agencies to monitor access and use. For managed-access data, the institution that manages data shall be responsible for data-audit. The data-management group shall regularly seek reports from users who have been provided data-access. Report of any breach in data-access or data-usage for open access or managed access data shall be appropriately dealt with by the national committee or the data-management group. Penalty for proven breach of access/use shall minimally comprise barring of access to the database for an extended period of time.
For publicly-funded projects, if data-generation is outsourced to any GoI or private agency, then a Data Delivery and Non-Retention Agreement must be signed by the agency to which data-generation is outsourced. This Agreement must clearly describe the nature of samples being sent to the agency, the data to be generated, the date of delivery of data. Further, the Non-Retention portion of the Agreement must state that (a) data more than what has been requested and paid for shall not be generated by the Agency, (b) quantities of samples remaining after data-generation shall be returned by the Agency to the P.I., and (c) no data shall be retained by the Agency after despatch to the P.I.
A model Data Delivery and Non-Retention Agreement form is provided in Appendix-4.
References: To be added
Examples of High-throughput Data Types
Data on imaging of cells, molecules, whole genome, whole exome, gene-panel, transcriptome, gene expression, chip-seq, methylome and other epigenome or epitranscriptome, metagenome, proteome, metabolome and any data with information on more than a single gene/protein/metabolite from microbes, plant or animal cells, cell lines, animal and human organs using multiple of platforms like next-generation sequencing, microarrays, mass-spectrometry and microscopy and bimolecular structures. Examples of some of the high-throughput data generated are data on genome-wide association study (GWAS), genome and exome sequencing, gene expression studies using RNA-seq or miRNA- seq, disease-specific gene panels, DNA microarrays, proteomics studies using mass spectrometry, imaging different types of cells in human body, understanding microbial population in human gut using metagenome sequencing and disease metabolite profiling using gas chromatography coupled with mass spectrometry (GC-MS) or liquid chromatography coupled with mass spectrometry (LC-MS).
Levels of data
Raw (Level 1) Data:
First level data converted from raw images. Examples of Level 1 data are FASTQ/CSFASTA/HDF5/SSF files, intensity (idat) files along with the manifest (bpm/bgx/TXT) file for Illumina microarrays, EXP and CEL files for Affymetrix arrays, dta/pkl/ms2/mgf files for protein mass spec data, spectrum data for metabolites, TIFF/JPG/PNG files for images, higher level coordinates for 3D structures for biological macromolecules.
Processed (Level 2) Data:
Raw (Level 1) data are curated, processed and analyzed to provide value-addition and to ease inferences. Examples of such data are BAM/CRAM/FAST5/ProBAM files for sequencing, nmrML files for metabolite profiling experiments, CHP file for Affymetrix microarrays, gtc files for Illumina microarrays.
Processing of raw or semi-processed data are done in a variety of ways. Examples of such higher-level processed data are VCF/BCF files for variants, TXT file with genes with analyzed expression values (FPKM/RPKM/TPM normalized BED files), BED/ProBED files for genomics and proteomics data respectively, SGA/GFF files for chip-seq experiments.
Data Request Form
1. Full Name of the Research Project for which Data are Requested
2. Applicant Information
Applicant (person in charge of the research project)
Name Telephone Work Address E-mail
Contact person (if other than applicant)
Name Telephone Work Address E-mail
3. Brief Description of the Research Project
Aims of the study, study design, scientific value and significance (max 300 words) Timetable of the study
Agency funding the study
4. Description of the Requested Data 5. Plan for Publishing the Results
Describe, how the results are planned to be published (e.g. scientific article, PhD thesis)
6. Signature (Principal Investigator)
Place and date Signature
Name in Capital Letters
|APVMA||Australian Pesticide and Veterinary Medicine Authority|
|BIS||Bureau of Indian Standards|
|BrDU||Bromodeoxyuridine / 5-bromo-2′-deoxyuridine|
|CIB||Central Insecticide Board|
|OECD||Organisation for Economic Co-operation and Development|
|EDX||Energy Dispersive X-Ray spectroscopy|
|FAO||Food and Agricultural Organization|
|ICP-MS||Inductively coupled plasma mass spectrometry|
|ISO||International Organization for Standardization|
|PLA||Polylactic acid or polylactide|
|PMRA||Pest Management Regulatory Agency|
|REACH||Registration, Evaluation, Authorisation and Restriction of Chemicals|
|TSCA||Toxic Substances Control Act|
|USFDA||United States Food and Drug Administration|
|USEPA||US Environmental Protection Agency|
|WHO||World Health Organization|
|WST-1||Water Soluble Tetrazolium Salts|
|XRD||X-ray powder diffraction|
Nanoparticles acquire unique properties due to their small size to large surface area ratio. It thus supports in development of novel products and processes as well as enhances the performance of existing ones across several disciplines. Nanotechnology has recently been introduced for improvement of agricultural systems through higher crop yield and better crop protection in order to meet the changing needs and domains of providing food to the growing population of the world. The innovative nano-intervention in agriculture and food sector could generate low-cost, high-efficacy solutions in terms of products and processes, especially suitable for developing countries. However, the unique properties can also lead to nanoparticle-related toxicity in humans and environment. The guidelines for evaluation of nanoproducts in agriculture and food are more challenging than the existing procedures for assessment of fertilizers or safety evaluation of pesticides or toxicity evaluation in food. The activity, efficacy and impact of nanomaterials (NMs) depend upon interaction of their physico-chemical parameters with diverse environmental factors and, therefore, require a multidisciplinary approach for development of new alternative strategies and methods for evaluation.
It is imperative to modify the existing policies and also develop certain new standard guidelines for evaluation of novel products on the basis of current scientific understanding. The multidisciplinary nature of nanotechnology and its rapidly increasing scope for development of commercially viable applications pose a huge challenge to regulatory bodies across the globe. Nanotechnology involves an amalgamation of knowledge from various disciplines of science, including chemistry, materials science, physics, biology, engineering and medicine. Such an interdisciplinary nature makes nanoscience an important domain to facilitate enhanced scientific and technological prospects and development of novel applications. Moreover, nanotechnology and nanoproducts are dealt by different ministries and different departments, and thus interdepartmental and inter-ministerial convergence is also required (Annexure 1). These guidelines have the aim to ensure not only the quality and efficacy to encourage the commercialization of nanotechnology-based innovations but also safety of novel products by emphasizing high benefit to low risk ratio compared to bulk counterparts.
There are no unanimously acceptable international guidelines for nano-agriproducts (NAPs). A few provisions that are in place globally for nanomaterials include REACH, EPA, AVMPA, OECD, and FAO/WHO with certain specific guidelines for quality, safety and efficacy. However, new innovations with alteration of functionality of nanosystems make it difficult to apply a universal set of evaluation parameters for different nanoproducts with different applications. Many a time the case-by-case basis evaluation approach is advocated for NAPs.
These guidelines apply to the following two categories of products:
These guidelines do not apply to the conventional products or formulations with incidental presence of natural nanomaterials. These guidelines also apply to sensors made from nanomaterials (as per the definition) and those that require direct contact with crops, food and feed for data acquisitions.
The European Union along with Switzerland is the only part of the world where particular provisions to deal with nanoproducts are available in the legislation. In some countries, in the absence of specific regulations for nanoproducts, the existing legislative and regulatory frameworks (Annexure 2) also deal with nanoproducts, many a time with necessary adaptations to account for the specific properties of nanomaterials. In India, there are different government bodies and provisions that regulate different agriproducts. Since different NAPs are considered in the guidelines, their evaluation should be conducted as per the NAP type. However, in case any specific study is not included in the suggested regulatory framework, the principles of ICH guidelines for agriproducts or OECD guidelines for chemicals may be fol- lowed. This document may also serve as useful guidelines for manufacturers, importers of NAPs and other stakeholders involved in research and development of NAPs.
The following nano-agriinproducts are considered in these guidelines:
(i) Nanofertilizers (with or without nanocarriers): Safety, evaluation, functionality and other quality studies of nanofertilizers should be conducted under Fertilizer (Control) Order (FCO), 1985 with additional criteria for inclusion of nanofertilizers. FCO is administered by Department of Agriculture Cooperation, Government of India and issued under the Essential Commodities Act, 1955, which lays down registration requirement for fertilizers.
(ii) Nanopesticides (with or without nanocarriers): Safety studies on chemistry, bio-efficacy and residues, toxicity, packaging and processing of molecules for registration for manufacture or import of nanopesticides should be conducted as per the regulatory aspect provisions under section 9(3) specified in the Insecticides Act, 1968 with additional criteria for inclusion of nanopesticides and plant growth regulators as per the guidelines of Central Insecticides Board & Registration Committee under the Ministry of Agriculture (Insecticide Act, 1968).
The following nano-agriproducts are considered in the guidelines:
DBT –DRAFT 1
(i) Nanofood: The FDA guidelines (FDA, 2014a), (FDA, 2014b), (FDA, 2015) and Food Safety and Standards Act, 2006 may be adopted by FSSAI (Food Safety and Standards Authority of India) to address NAPs and develop final guidelines for industry. In these guidelines, NAPs are classified according to their degradability, organicity, function, approvals and how they have been synthesized. Accordingly, the safety and efficacy data requirements are described.
This document may serve as useful guidelines for manufacturers, importers of NAPs and other stakeholders involved in research and development of NAPs. These guidelines are aligned with the provisions of REACH, OECD and FAO/WHO with certain specific aspects of quality, safety and efficacy applicable to nano-agriproducts. Specific scientific evidence is required for approval as per approaches for evaluation of such products that has been covered under this guideline. Each application should be considered on its own merit of the data submitted using scientific evaluation and valid justification.
(ii) Nanofeed: Safety, evaluation and other quality studies of nanofeed should be conducted under Cattle Feed (Regulation of manufacture and sale) Order, 2009 with additional criteria for inclusion of nanofeed.
A NAIP is defined as an agricultural input preparation containing nanomaterials intended for external and internal applications (through soil, seed, foliar, and drip in crops as well as by other means) on crop for the purpose of agricultural farming.
NAIPs consist of materials with any of the three dimensions, that is, zero, one or two, on the nanoscale or with an internal or surface structure in the nanoscale. The nanomaterial is defined as a material that ranges in size from 1 to 100 nm at least in one dimension. However, if the particle size is >100 nm and <1000 nm, it may also fall within the definition, provided it has altered the agri-input product characteristics associated with the application of nanotechnology compared with active ingredient. The variations in definition of nanomaterials with respect to size in different countries and respective regulatory bodies are presented in Annexure 3.
A NAP is defined as an agricultural preparation containing nanomaterials intended for consumption or applications in food/feed and their supplements as well as nutraceutical delivery. These are the products that contain materials with any of the dimensions (zero, one or two) falling under the size range of >100 nm and <1000 nm, provided the particle size has altered the agriproduct characteristics associated with the application of nanotechnology compared with the active ingredient.
NAIPs could be categorized depending on the properties and functionalities of NMs and the existing products containing synthesized and engineered NMs. Complete categorization scheme of nanomaterials is given in Annexure 4. NAIPs could be categorized as follows:
(i) According to degradation nature of nanomaterial
DBT –DRAFT 1
• Biologically synthesized NMs: Nanomaterials that are synthesized using bio-agents and their bio-actives. Examples include metallic NPs, bimetallic NPs, metal oxide NPs, quantum dots, nanoclusters and reduced graphene.
DBT –DRAFT 1
The rationale underlying manufacturing of NAIPs and NAPs should be demonstrated and specified with reference to their claimed advantage in comparison to conventional products. The NMs and their transformed waste disposal may have an adverse impact on the ecosystem. Therefore, the known and supposed adverse impacts on ecosystem should also be taken into consideration. The following aspects should be specifically addressed for justification of the use of NAIPs and NAPs:
These guidelines are developed in provisions of Insecticides Act, FCO, BIS and FSSAI, with specific requirements and adaptations for NAIPs and NAPs wherever considered necessary. While these provisions specify the general requirements and guidelines to manufacture or import new fertilizers (FCO), pesticides (Insecticide Act), food additives and preservatives (FSSAI) or to undertake quality checks, this document provides guidance on specific requirements for agri-input and agriproducts developed based on nanotechnology. General requirements as specified in these provisions will be applicable for any new products whether nanotechnology based or not. However, because of the involvement of interdisciplinary sciences and the complex nature of NAPs, a ‘case-by-case basis’ approach should be adopted for their evaluation with respect to enhanced efficacy and safety.
Considering the unique process conditions of nanoformulations compared to the conventional agri-input products and agriproducts, the product description should include detailed description methods of manufacturing process (excluding critical intellectual property information) and process controls to be included. The method of nanoparticle waste disposal and environmental impact may be declared.
‘Nanocomponents’ incorporated into some specific materials such as plastic, ceramic and regenerated cellulose films are subject to different kinds of regulations. The policy 2002/72/EC (14) implemented in Great Britain may be followed to regulate plastic and other food contact articles to deal with food contamination issues due to migration of lead and cadmium. European Regulation No. (EC) 1935/2004 may be followed to evaluate quality and safety of foodstuffs. The 12 principles of green chemistry proposed by EPA in 1991 may also provide guidance for engineering safe NAIP and NAPs. These include prevention, atom economy, less hazardous chemical syntheses, designing safer chemicals, safer solvents and auxiliaries, design for energy efficiency, use of renewable feedstocks, reduce derivatives, catalysis, design for degradation, real-time analysis for pollution prevention and inherently safer chemistry for accident prevention.
Excipients help in the manufacture of formulations of NAIP and NAPs and improve performance and stability of the product. Examples of excipients in NAIPs and NAPs include stabilizers to prevent agglomeration and aggregation, preservatives to prevent microbial growth, surfactants and coupling agents to modify surface characteristics of nanomaterial.
The general storage stability requirements and procedures for agricultural chemical products may also be applied on NAIPs and NAPs to ensure stability. The following four principal types of storage stability studies as per OECD TG 318, FAO/WHO, AVMPA may be adopted:
The test parameters for stability testing of NAIPs and NAPs are also considered. The following test parameters (whichever is applicable) may be considered for each product. Relevant scientific argument should be provided to explain why to exclude any one of the following test parameters:
dispersion stability, dilution stability, particle size distribution, emulsifiability, re-emulsifiability, emulsion stability, viscosity, flowability, crystalline state, release kinetics and leakage).
(g) Microbial stability
Depending upon the product type, application and exposure to humans and environment, the suitable in vitro methods for hazard assessment and effective regulation of NAIPs and NAPs should be adopted from the listed items. Each of the in vitro assays mentioned is based on existing OECD Testing Guidelines (TGs) for application to testing manufactured NMs (OECD, 2019):
Dermal exposure/toxicity: OECD TG 428 (in vitro); OECD TG 427 in vitro kin corrosion: OECD TG 431 Eye irritation: OECD TG 437
Genotoxicity: OECD TG 471, 473, 476, 482 and 487
Inhalation exposure (toxicity): OECD TG 403
Cytotoxicity assays used in the OECD testing programme: ATP CellTiter-Glo, neutral red uptake, LDH release, MTT, XTT, cell impedance, trypan blue, BrdU, Alamar Blue, WST-1, live/dead cell counting, colony forming efficiency, genotoxicity assays used in the OECD Testing Programme: Comet assay and DNA double-strands breaks
Information on the persistence of NAIPs and NAPs should be provided in the registration dossier and residues analysis of used nano active ingredients, additives, and nanocarrier materials needs to be performed during the life cycle of the product. Data on nanomaterial residue, presence of any nanosized degradation products in food/feed, excipients or surface coating used on food contact material need to be declared by the manufacturer during product registration. The report must mention the following details:
Method for determination (detection and quantitation limits) of residues from the used active ingredient, additive and nanocarrier
Quantities of generated residues and summary of anticipated risks of generated residues: The requirement for toxicological data, there is no migration of elements from food contact materials or the migrating species are not in the nanomaterial form (in which case standard risk assessment should apply)
Information required for evaluation of NAIPs
Nanopesticides (chemical and biological sources) and growth regulators are required to be registered under the existing Insecticides Act, 1968 and Rules 1972 through nodal agencies, namely, Central Insecticides Board and Registration Committee. Likewise major fertilizers (chemical and biological sources) and micronutrients are required to be registered under the existing FCO, 1985 through the nodal agency Department of Fertilizers, Government of India and agriculture department of state governments. The food and feed products developed using nanotechnology interventions are required to be registered as per the guidelines by FSSAI. The nanotechnology intervention used in these products must be registered under the existing regulations. The additional data sets required for registration of nano-agriproducts based on the active ingredient and nanocarrier formulated products are as follows:
o Nanopesticides (insecticide, fungicide, nematicides, acaricide and rodenticide)
Information on the ingredients
Information on nanomaterials used (active ingredient/nanocarrier)
Used nanomaterials based on the method of production and composition Nanomaterials property characterization
Hydrodynamic particle size and distribution (polydispersion index)
Surface charge (using zeta potential)
Transmission electron microscopy (for shape, size and actual average particle size) Aspect ratio (only for 1D and 2D nanomaterials using TEM, SEM and FE-SEM) Hydrophilicity/lipophilicity using contact angle measurement
pH using pH meter
Viscosity (in case of liquid formulation using viscometer)
Electrical conductivity (in case of liquid formulation using conductivity meter) Organic (HPLC and GC data); inorganic (XRF and ICP-MS data)
X-ray diffraction chromatogram
Stability data (as per OECD 318 TG) Impurities detail
Quality control checks parameters and test protocols Sampling procedure and preparation for specific analysis Testing protocol/s
Certificate of analysis
Preliminary toxicity analysis data (confirmatory toxicity analysis would be performed by CIB)
b. c. d.
Cytotoxicity: ATP Cell Titer-Glo, neutral red uptake, LDH release, MTT, XTT, cell impedance, trypan blue, BrdU, Alamar Blue, WST-1, live/dead cell counting, colony forming efficiency Genotoxicity: OECD TG 471, 473, 476, 482 and 487
Decision framework (OECD, ENV/JM/MONO(2019)12) for inclusion of physico-chemical parameters for exposure and fate assessment of nanofertilizers and nanopesticides may be followed.
In addition to the information mentioned in Section 12, the information discussed next will also be required for NAPs.
13.1 Exposure risk
NM exposure measurement is essential for hazard characterization and risk assessment. Migration of NMs or its degraded products in non-nano form (its type and quantity) from agri produce or via animals for food production or from food contact materials (like packaging) should be considered in exposure measurement and hazard characterization and ADME studies are required. Specific testing protocols for analysis of migrated products are required. Food sampling, variability in composite sampling and concentration variations between samples are critical sampling issues in exposure evaluation. The decision framework defined by OECD (ENV/JM/MONO (2019)12) may be followed for inclusion of physico-chemical parameters for exposure and fate assessment of nanofertilizer, nanopesticide and NMs in food. Guidance on risk assessment of the application of nanoscience and nanotechnologies in the food and feed chain (EFSA, 2018) may also be followed.
13.2 Hazard characterization
Hazard identification and characterization require appropriate in vitro and in vivo studies to determine the fate of NMs. Toxicity testing should be customized for case of exposure.
1. In case of the non-stable NMs in food preparation/formulation: For example, when NMs are completely degraded/solubilized/transformed to their non-nano form in food matrices, general protocol for toxicity measurement of non-nano form in the intended application can be considered. But strong scientific evidence should be produced demonstrating its solubility. This criterion applies to non-persistent NMs in marketed foods and foods where nano form transforms to non-nano form before injection.
2. NMs that get transformed during digestion: For NMs that get completely degraded/dissolved in gastrointestinal tract and where there is no possibility of their absorption in nano state, the hazard characterization can be relaxed and can rely only on data for non-nano form. This scenario should be strongly supported by in vitro genotoxicity and in vivo testing for local effects and other in vivo tests. When regulations on non-nano form are not available, FSSAI has to come up with regulations.
3. Stable nano materials: For the nanomaterials that are stable in food formulations/agri produces and
in gastrointestinal tract, two scenarios considered:
4. Migration of food contact materials: When there is no migration from FCM ,toxicological concerns are negligible. If not, stringent toxicity studies need to be enforced.
The following types of toxicity testings are required for NMs (EFSA Scientific Committee, 2011):
I. In vitro studies: They help to understand biological responses of NMs and underlying mechanism for toxicity screening. However, suitability of test system and possible structural and functional changes arising from interaction of NMs with culture medium should be considered.
II. In vivo studies: In vivo studies are essential to identify ADME profile, adverse responses and dose- dependent toxicity. Forms of administration of NMs (e.g., adding to feed, water or by gavage) for in vivo studies also influence the toxicity profiling. For example, NMs that interact with food and form complex matrices, simulant cannot be used and it should be homogeneously blended in food.
OECD test guidelines 414, 415 and 416 may be adopted for study design of reproduction and developmental studies.
13.4. Uncertainty analysis
Analysing possibility of uncertainty in assessing the above-mentioned assessments. Some of the possible reasons for uncertainty in assessing ENM are as follows:
Hundreds of NAIPs and NAPs using nanotechnology are already on the market even though there are no specific policies regulations for their control. Therefore, there is a need to develop the policy and regulations in place.
DBT –DRAFT 1
J Food Sci 75(9):E627–E635
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We invite companies who operate in Biostimulants & Biological products for Agriculture sector. (No pharmaceutical, vet or any allied sectors) to join hands with us to take this initiative forward and jointly raise voice against common issues faced for the segment.