Project-wise Reports of Sectorial Monitoring Committee

1. Project Title: Processes and Products from Himalayan Region and their Toxicological Evaluation (PROMOTE} BSC-0213

2. Project Details:

Nodal lab CSIR- Institute of Himalayan Bioresource Technology

Participating Labs NA

Nodal Officer Dr. Bikram Singh

Taskforce 1st Taskforce meeting (Director, CSIR-1 HBT} Chairman 2nd Taskforce meeting (Director, CSIR-IHBT)

3rd Taskforce meeting (Working Director, CSIR-IHBT} 4th Taskforce meeting (Dr. O.P. Sharma} Opted Chairman 5th Taskforce meeting (Director, CSIR-IHBT)

Total Man Days 3185 man days

Approved Cost 1127.450 (Lakhs}

Fund received 894.320 (Lakhs)

Fund utilized 871.800 (Lakhs}

3. Periodic Key Recommendations of Task Force/RC/Monitoring Committee

Annexure 1

S.No Key Recommendations Action taken on Key Proposed of Task Force/RC/ Recommendations Deliverables Monitoring Committee

4. Project Achievements:

s. Deliverables promised at Deliverables Achieved No. start

WP-1 Quality improvement of Steviol glycoside. 1 Increased quality Process for extracting SGs with purity of more

enrichment of SGs on than 90% SGs standardized with reduced water small/ laboratory scale. consumption. (Achieved) Process & equipment Process developed at pilot scale and up scaled design, Upscaling of upto 40 kg per batch dry stevia leaves for processes developed on purification of SGs at pilot scale. (Achieved) laboratory scale and Pilot scale purification of SGs. Feasibility studies and Achieved optimization of process parameters. Validation of processes and Validation is under progress with raw stevia

Revised Deliverable (approved by Task Force)

If not achieved, reasons thereof

Quality evaluation of SGs leaves from Punjab. (Partially in process) (including physical and chemical)

WP-2 Process development for natural colours and dyes. 1 Up-scaling of processing (50 kg fresh batch capacity)

technology from lab to pilot Processed around scale and process feasibility 2 tons black carrot for extraction of natural colour studies. (Achieved)

2 Stability evaluation of The stability was evaluated by using different natural colours and dyes parameters. Similarly, the tech no-echom ic Techno-economic evaluation has also been carried out. (Achieved) evaluation of process

3 Exploring the waste Extruded snacks product standardized using streams for production of dried black carrot pomace. (Achieved) value added products

WP-3 UpscalinQ of theaflavinproduction. 1 Process for theanine Process for theanine production at lab scale.

production at lab scale. (Achieved)

2 Process for theaflavin production at pilot scale. I. Process for catechin Process for catechin production at pilot scale.

production at pilot scale. (Achieved)

II. Variations in PPO Variations in PPO activity in the tea clones at activity in the tea clones IHBT Tea Experimental farms was studied.

(Achieved)

Ill. Process for theaflavin Process for theaflavin production from catechins

production from purified is standardized at lab scale. (Partially in

catechins progress.)

WP-4 Development of functional food products. 1 Local bioresources will be Local bioresources such as Pomegranate (Daru),

evaluated for their Murraya K and Stevia has been selected. nutritional profile. (Achieved)

2 Selected plant/process/ Nutritional and functional properties has been food component will be accessed. (Achieved) used for nutritional enrichment.

3 Development of suitable Product formulation has been achieved at lab process methodologies for scale level. (Achieved) various innovative food products.

4 Optimization of extracting Optimization of extraction for functional techniques with respect to component has been done at lab scale level. purified food, yields and (Achieved) functional ity.

5 Up Scaling of processes for Up scaling of processing for preparation of preparation of final optimized products are in progress. (Partially in formulated foods. progress)

WP-6 Development of biopesticides. 1 A. Up-scaling and Produced Helicoverpa armigera nucleopolyhedro-

2

formulation development of nucleopolyhedro­virus (NPV) lead for the management of Helicoverpa armigera

2 Identification and molecular characterization of efficacious nucleopolyhedrovirus isolates for the control of Spodopteralitura

virus (HaNPV) polyhedral inclusion bodies (PISs) by infecting H. armigera larvae for development of large quantities of formulations. The produced PISs are enough for spraying 2 ha of crops Standardized formulation recipes of Helicoverpa armigera nucleopoly-hedrovirus (HaNPV)

Director of Extension Education, CSKHP Agriculture University, Palampur has agreed to conduct field efficacy trials of the developed formulations against H. armigera infecting peas and tomatoes. Trials will be conducted from Jan­April 2017 (Achieved) Collected diseased larvae of Spodoptera litura showing viral like symptoms for the identification and molecular characterization of nucleopolyhedro-virus. (Achieved)

B. Development biopesticides

of Identified thirteen entomopathogenic fungi (EPF) native strains from insect cadavers and soil.

Two native strains of EPF showed bio-efficacy against aphids and diamondback moth under laboratory and greenhouse conditions. (Achieved)

2 The five new strains (Strain B, C, D, F and H) identified showed excellent activity against second instar larvae of P. xylostella at 1x109 spores/ml (93-1 00% mortality) and was followed by 1 x1 os spores/ml (76-86% mortality). (Achieved)

WP-7 Bioprospecting abundantly available Himalayan flora 1 Identification of bioactive Isolation and identification of eight alkaloids from

compounds for the Adhatoda vascia including vascine, which is preparation of health care developed as organocatalyst for synthesis of products /drugs various drug intermediates.

Isolation and characterization of fifteen molecules from Zanthoxylum armatum. The molecules were evaluated for in vitro activity against different cell lines. The enriched fractions having around 0.1% of lignans, flavonoids also having 0.1% were processed. Value addition of parthenin isolated from Parthenium hysterophorus was carried out to prepare different analogues and the insecticidal activity was also evaluated which was found to be active at higher concentrations. found a potential source of shikimic acid which is important starting product for the production of drug Oseltamivir. (Achieved)

WP-8 Development of methodologies for synthesis of different classes of triketones for herbicidal activities. 1 Application of 4-substituted Four triketo compounds (i). 4-methyl

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cyclohexane-1 ,3-diones for cyclohexane-1 ,3-dione, (ii). 4-isopropyl triketones synthesis. cyclohexane-1,3-dione, (i ii). 5-methyl

cyclohexane-1,3-dione and (iv). 5-phenyl cyclohexane-1 ,3-dione) have been synthesized and tested for field level non selective herbecidal activity but need more efforts to find out active molecules and better solubility. (Achieved)

2 Evaluation of synthesized Preliminary field level studies have been done but triketones for herbicidal need more modification in structure and trial to activity. reach up to active herbicide (Achieved)

WP-9 Development of hybrid nanocomposite. 1 Protocol for synthesis of Standardized chemical synthetic process for the

hybrid nanocomposite preparation of hybrid nanocomposite (Achieved)

1- 2 Functionality of hybrid Bioconjugation with the hybrid nanocomposite nanocomposite for have been achieved using amide coupling multimodal imaging reaction (Achieved)

3 Toxicity testing for in vitro Cytocompatibilty studies with two different and in vivo level samples at various concentrations have been bioaccumulation done. (Achieved)

4 Selection of safe hybrid Work is in progress nanocomposite for use

WP-1 Oin vitro production of secondary metabolite from Arnebia 1 Optimization of condition Shake flask optimization work under progress,

for production of shikonin in which later can be used for scale up in bioreactor bioreactor

WP-11 Toxicity testing and evaluation of value added products developed by CSIR-IHBT. 1 Toxicity and efficacy testing Acute and sub-acute toxicity testing of steviol

of developed products glycosides. (Achieved) under different work Safety studies anthocyanin-rich black carrot packages extract. (Achieved)

Acute oral toxicity study of Pomegranate granatumpeel extract. (Achieved) Cardioprotective effect of Pomegranate granatum peel extract. (Achieved) In vitro and in vivo testing of hybrid nano composites. (Achieved)

5. Outcomes and outputs from the projects:

a. Outcomes

1. Lead(s) identified : Number and names with TRL

Number: Nine (10)

I. Process for extraction of steviol glycoside with purity of more than 90% SGs from Stevia rebaudiana leaves (TRL: 6)

11. Standardization of technology for production of natural colours employing green technology at pilot scale (TRL:6)

Ill . Process for extraction and purification of catechins from tea shoots (TRL:6) IV. Healthy extruded snacks incorporating pomegranate peel developed. (TRL: 5) V. Ready to use stevia pouches, liquid drops and tablets developed (TRL: 6)

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VI. Identified efficacious isolate of nucleopolyhedrovirus (NPV) for management of Helicoverpa armigera (TRL:4)

VII. Identified native strains of entomopathogenic fungi (EPF) for the management of diamondback moth (Piutella xylostella) in crucifer crops and aphid (Aphis craccivora) in agricultural/horticultural crops (TRL:4)

VIII. Identified a potential source of shikimic acid which is important starting product for the production of drug Oseltamivir phosphate (TRL: 4)

IX. Scalable process for substituted cyclohexane-1 ,3-diones as precursor of herbicide (TRL: 6) X. Synthesis and bio conjugation of fluorescent nanocomposite as nanodiagnostic probe (TRL: 3)

2. Technology/Product developed: Number and names with TRL level of Development

Four (4)

2.1. WP-1: Steviol glycosides with purity more than 90% and its formulated products (TRL6}

4 stakeholders identi fied for technology transfer: • M/s Agri Natural India, Ludhiana • M/s BLK, Bengaluru • M/s Himalayan Product, Lohna, Palampur • Mr. Virender Singh, Bulandsehar

2.2. WP-2: Extraction process for natural colours from black carrot (TRL6)

3 stakeholders identified for technology transfer: • M/s. Aarkay Food Products Ltd. Ahmadabad, Gujarat • M/s. Arun Colour Chern. Pvt. Ltd., Delhi • M/s. Ajanta Colour Chern. Pvt. Ltd. , Delhi

2.3. WP-3: Process for catechin production (TRL6)

2 stakeholders identified for technology transfer: • Technology transferred to M/s. Baijnath Pharmaceuticals Pvt. Ltd., Paprola, HP • INDCOSERVE, MSME, Tamil Nadu identified as potential stakeholder for transfer of

technology

2.4. WP-4: Development of functional food products (TRL6)

• Stakeholder for stevia tablets: M/s Himalayan Product, Lohna, Palampur

2.5. WP-6: Development of biopesticides (TRL4) • Stakeholder:

Multiplex Bioteck Pvt. Ltd. P. J . Margo Pvt. Ltd. Banglore

2.6. WP-7: Bioprospecting aboundantly available Himalayan flora (TRL: 4)

3. Technology/Product transferred: Number and names with name of licensee(s) Three (3)

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3.1. M/s Agri Natural India, Ludhiana for stevia production from Stevia rebaudiana.

3.2 MoU signed for making stevia tablets with one incubatee entrepreneur M/s Himalaya Natural & Herbal Products on actual basis of expenditure at CSIR-IHBT, Palampur.

3.3. M/s. Baijnath Pharmaceuticals Pvt. Ltd., Paprola for catechin production from tea shoots

4. Facility created: Number and names with uniqueness & utility: 2 No. (costing Rs. 2,02,20,000) • lon exchange and adsorption column (Rs. 22,00,000)

This column can be operated in co-current and counter current mode and prior to commercialization adsorption studies can be used to further upscale the process.

• Image stream, flow cytometer (Rs. 1 ,80,20,000) This instrument is useful for cell phenotyping and imaging of cell population under analysis.

3. Sponsored projects obtained (with details):

• BIRAC, DBT (Rs. 17.5 Lakhs) • Fast Track Translation (FTT) project (Rs 140 Lakhs)

4. External Cash Flow (ECF) generated: Rs 26.31 Lakhs

5. Societal impact created

Products developed under the project were demonstrated during India International Trade Fair 2016 and Indian Science Festival 2016 at New Delhi, where more than 200 progressive entrepreneurs showed interest in our technologies and products.

• Natural and safe sweetener is the need of the hour with the world population showing an increase in number of diabetics. Steviol glycosides are safe, natural non-calorific sweeteners. This product will give consumers a safe alternative to sugar and artificial synthetic sweeteners such as aspartame, sucralose etc. A successful stevia industry in India could provide following societal impact /benefits as fo llows: For health of public by consuming low calorie healthy natural sweetener. To growers, enabling greater diversification opportunity along with additional employment if a commercial extraction plant is established.

• Tea industry is faced with labour shortage which affects the quality of the tea. The developed process for extraction of catechins employs the tea shoots which are not fit for making tea for commerce. Catechins are high valued tea phenolics with reported health benefits. The extraction of catechins from the shoots instead of manufacturing tea would value add the low-quality overgrown shoots. Catechins have a wide market with applications in food, beverage, cosmetics and nutraceutical industry and fetch good price.

• Use of synthetic colours and dyes has been reported to have many adverse health effects like cancer, etc. and have been banned for use in food and beverages by a number of countries. Natural colours and dyes are the safe alternatives to these synthetic colours and dyes. The commercialization will be beneficial for health of general public by consuming safe colours in foods and beverages, confectionaries etc. To growers, for better returns on their produce along with additional employment if a commercial extraction plant is established.

b. Outputs:

1. Periodic Reports Submitted 6

s. No.

1

Quarterly, half yearly and yearly reports from 2013-2016)

Minutes of the task force meeting held on 5 Sept. 2013, 27 Feb. 2014, 4 Sept. 2014, 31 March 2015, 7 Oct. 2015

RC Evaluation Reports and Minutes

2. Publications i. Total number: 25 ii. Cumulative IF (as per 2015 figures) : 38.11 iii. Average IF: 1.65 iv. Average IF/scientist: 2.38

3. Patents (or other forms of IP generated such as Copyrights, Designs etc.) i. Filed/ Granted

Filed /granted: 2

3.1. WP-9: Acharya, A "A novel hy!brid nanocomposite material for Opticai/MRI bimodal molecular imaging, and a process for the preparation thereof' Patent No. IN 2014DE01629.

3.3. WP8: Pralay Das, Dharminder Sharma, Bikram Singh, "Substituted cyclohexane- 1,3-dione compounds, process for preparation thereof and its applications", US 8,916,723 82 (date of patent Dec. 23, 2014}.

ii. Licensed: Nil

4. Human Resource generated

i. Ph.D. fe llows (give PhDs completed & pursuing separately): 5 No. (2 completed & 3 pursuing)

ii. Project fellows: 26 iii. Skills imparted

- Including skill set developed for technical services/ consultancy/ training purposes • Imparted orientation-training programme to 30 students of National Institute of Fashion

Technology, Kangra, H.P on production of natural colour at pilot plant. • Imparted training to 5 technical persons of M/s. Baijnath Pharmaceuticals Pvt. Ltd., Paprola

on production of catechins from tea shoots • Trained 3 technical persons for stevia extraction from Agri Natural Ludhiana • Products demonstrated during India International Trade Fair 2016 and Indian Science

Festivals 2016 at New Delhi, where more than 200 progressive entrepreneurs shared interest in our technologies and products

5. External collaborations established: CSKHPKVV on field trials for bio pesticides

LeadfTechnology/Pr Salient TRL of the IP Status of Status oduct (listed above Lead/ Lead/Tech nology/Pr DPR/Commercial

of

in outcomes) Features of the

Technology oduct Technical Report of Lead/Tech nolo /Product Lead/ Technology/ gy/ Product Product

Steviol glycosides Green water TRL: 6 Part of the process Will be ready by April with purity more than based extraction patented in XI FYP. 2017 90% and its and purification Rest of the novel formulated products technology steps are under

preparation.

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2

3

4

5

6

Process for extraction and purification of catechins from tea shoots

a. Healthy extruded snacks incorporating pomegranate peel developed

Green and does not employ any organic and harmful solvents

Low cost and easily adaptable technology. Health and nutritional components of

b. Ready to use ingredients stevia pouches, liquid preserved in the drops and tablets products developed

TRL: 6

TRL: 6

a. Identified Novel strains TRL: 4 efficacious isolate of nucleopolyhedrovirus (NPV) for management of Helicoverpa armigera b. Identified native strains of entomopathogenic fung i (EPF) for the management of diamondback moth (Piutella xylostella) in crucifer crops and aphid (Aphis craccivora) in agricultural/horticultu ral crops

identi fied

Identified a potential Potential TRL: 4 source of shikimic resource in acid which is Himalayan plant important starting identified product for the production of drug Oseltamivir phosphate

Scalable process for substituted cyclohexane-1 ,3-diones as precursor of herbicide

Novel scalable TRL: 6 process developed for substituted cyclohexane-1,3-diones

Process already Will be ready by patented in the XI December 2017 FYP

Patent preparation

under Will be ready by June 2017

Leads will be Will be ready by transferred to December 2017 biopesticide industries for commercial application in agriculture for pest management

Process for the By the end of 2019 extraction and production of shikimic acid will be developed for commercia l venture

Pralay Dharminder Bikram

Das, Will be ready by Sharma, December 2017

Singh, "Substituted cyclohexane- 1 ,3-dione compounds, process for preparation thereof and its applications", us 8,916,723 82 (date of patent Dec. 23, 2014).

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6. Sectoral Monitoring Committee Recommendations on Lead/Technology/Product (listed above}:

For each Lead/Technology/Product provide details separately:

Criteria SMC Recommendations

Name of Lead/Technology /Product • Upscaling up-to 1 ton per batch for steviol glycoside . • Enrichment of rebaudioside A • Quality Improvement - catechin production • Ready to use stevia pouches, liquid drops and tablets • Health Extruded Snacks incorporated with pomegranate peel • The process for the extraction and production of shikimic acid • Scalable process for triketo herbicide synthesis

Is the Lead/ Technology/Product Yes worth taking up further?

What is the further R&D efforts that Scale up and optimization of processes for commercialization is need to be put by CSIRICSIR required for the above leads. The availability of raw material may laboratories? be checked.

Global benchmarking of the The products that are being developed are for Indian society and Lead/Technology/Product made affordable. Publication in journals of international repute specifically wrt specifications and cost

Is the Lead!T ech nology/Product No worthy of commissioning a DPR at this stage?

What are the likely resources and Financial support would be required for 3+2 years. time duration required for taking forward the identified Lead/ Technology/Product to the desired TRL?

Potential stake holders who may be Farmers, Industries and Indian society at large. appropriate to partner technically as well as financially

Suggestion for plausible road map Details would be mentioned once project proposal in detail would towards further development of be presented lead/technology/product for achieving desired TRL

GO/NO GO further development Go

Other suggestions, if any, related to Discuss with other institutions working on stevia and ocimum benefits/usage/ commercialization

7. Identified "lessons to be learnt", especially from shortcomings/failures.

Greater programmatic alignment would further strengthen the likely outcomes and impacts.

8. Comments on financial progress on the project • fund availability and utilization of fund in the project:

The financial progress is satisfactory and the investment made is justified.

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9. Sectoral Monitoring Committee Recommendations on Facility creation or other outcomes:

Experiences gained, materials generated and lessons learnt are expected to have far reaching impacts in years to come.

10. Grading of Project Execution: Outstanding/ExcellenWery Good/Good/Satisfactory/Un-satisfactory

Very good

11. Additional comments, if any:

None

~ Dr. Sudeep Kumar

(Member)

Dr. B. Sasikeran (Member)

Dr. Prosenjit Bose (Member)

Direct r, CSIR-CFTRI (TF Chairman · Member)

~~ Dr. R.S. Sangwan (Member)

Prof. Akhilesh Tyagi (Member)

Dr. Sudhir Achar (Member)

D~IR-CIMAP (TF Chairman - Member)

Dr. Suresh Ramamurthi (Member)

Or. Jr;£;· (Member)

Director, CSIR-CCMB (TF Chairman - Member)

S7~ Director, CSIR-IHBT

(TF Chairman · Member)

~,M}-Director, CSIR-IICT (TF Chairman - Member)

Director, CSIR-NBRI (TF Chairman - Member)

Director, CSIR-NIIST (TF Chairman - Member)

Director, CSIR- NCL (TF Chairman -Member)

sl::! (Member)

~ Dr. Arjula Ramachandra Reddy

(Member)

Prof. N rpinder Singh (Member)

Jll!fl--Dr. Mangala Rai

(Chairman)

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