Introduction & Overview

Mass spectrometry (MS) is a powerful analytical technique that is used to determine the molecular mass of molecules by measuring the mass-to-charge (m/z) ratio of respective ions. The MS principle consists of ionizing neutral molecules to generate charged molecules or fragment ions which are further separated according to their m/z values and finally detected. The application of mass spectrometry to drugs, synthetic organic molecules, active pharmaceutical ingredients (APIs), natural products, peptides, proteins, carbohydrates, nucleic acids etc are well known.

The hyphenation of liquid chromatography (LC), photodiode-array UV-Vis detector with the atmospheric pressure ionization mass spectrometry (APIMS) has proven to be an efficient on-line system for establishing the identity of organic compounds in any given test samples. Broad range of molecules such as volatile, non-volatile, ionic, etc. can be analyzed at the expense of very low amount of sample. High resolution mass spectrometer with tandem mass spectrometric (MS/MS) analysis is proficient to identify unknown compounds, to quantify known compounds and to elucidate the structure of molecules. Mass Spectrometry Facility at CDRI consists of state-of-the-art mass spectrometers that include ESI/APCI quad, ESI/APCI ion trap and Orbitrap MS.

Capabilities, expertise and services offered

Small molecules analysis:

• ESI/APCI-MS analysis.
• Qualitative LC/MS method development and analysis.
• Quantitative LC/MS method development, validation and analysis.
• High resolution mass spectral analysis (HRMS)

 Trainings:

We offer training programs to motivate interested postgraduate students and research fellows with an opportunity to spend valuable time in gaining practical experience in the field of mass spectrometry. We offer three different time-bound training programs given below: Course fee may vary based on duration and technique

S. No.	Training on	Duration
1.	LC-MS data interpretation	1 week
2.	LC-MS/MS data interpretation	1 week
3.	LC-MS & LC-MS/MS Sample preparation, qualitative, quantitative analysis and data interpretation	3 months

 Workshops:

Awareness workshops are being held every year in CSIR-CDRI, Lucknow to keep the users informed about the scientific advancements in these areas. This programme also provides a platform for younger generation to explore the application of sophisticated instruments for their research work.

 Sponsored projects:

• We take-up sponsored projects to analyze small molecules, present in the complex matrices, qualitative and quantitative analysis of targeted compounds (phytopharmaceuticals), generation of mass spectrum fingerprint of herbal products. Standardization of Herbal Medicines (HMs), MS based phytochemical analysis and quantification of phytochemicals for marker identification, quality control, authentication of raw plants and their formulations.

 Open access facility:

• TMS database (http://tmsdatabase.org/) is a library of MS/MS spectrum of naturally occurring compounds (Natural products). It is Designed and Developed by CSIR - Central Drug Research Institute, India.

The general approach for search are

General Search: by Name, Formula, CAS Number and Molecular weight. Mass Spectrometry Based Search: By MS/MS, ion search or product ion search for single compounds, advanced search and comprehensive search included multiple searches or for Mass spectrometry LC-MS/MS, data based search.

Digital library of Indian medicinal plants and their metabolites

The hyphenated MS techniques not only report the precise mass of compounds but also generate fragment ion spectra for identification /confirmation of compounds. This fragmentation pattern offers a unique molecular signature of organic compounds. As a result, web-based LC-MS/MS library (http://plantmetabolome.cdri.res.in/) of plant secondary metabolites with suitable software search algorithm serves the purpose of inverse search (distribution of known bioactive compounds in different plant species).

On the basis of above facts CSIR-CDRI, Lucknow developed a prototype of bioinformatics product to explore chemical diversity of Indian medicinal plant.

The present development (Digital library of Indian medicinal plants and their metabolites: a bioinformatics tool) provides mass spectrometry based methodologies to explore the chemistry of medicinal plants. It has targeted applications for medicinal plants chemistry.

1) Identification and authentication of plant species based on their Mass Spectrum/HPLC/LC-MS Fingerprints.
2) Discovery of known bioactive compounds from previously unknown sources (inverse search).
3) Distribution of known compounds in different biological source (Plants/herbs).
4) Identification of known/unknown derivatives of bioactive compounds in different biological source (Plants/herbs) using LC-MS/MS data.

Major Equipment and Instrumentation

1. Thermo Scientific Orbitrap Velos ProTMHyphenated with Thermo Scietific Surveyor HPLC system. Ionization Mode: ESI and APCI Mass accuracy: m/z 50-2000 and m/z 200-4000 Mass Resolution: > 100,000 at m/z 400 Mass accuracy: < 3 ppm RMS with external calibration and < 1 ppm RMS using internal calibration Multiple fragmentation techniques: CID and HCD, Parallel MS and MSn analysis
2. Waters Xevo TQD triple Quadrupole mass spectrometer Ionization Mode: ESI and APCI Mass Range:m/z 5-2000 Th Mass Resolution:Unit Mass (Da), Fullscan MS and MS/MS, SIR & MRM
3. AB Sciex 4000 Q-Trap-Mass Spectrometer, Hyphenated with a Waters Acquity UPLC™ system. Ionization Mode: ESI and APCI, Mass Range:m/z 3-2800 (quadrupole mode) and m/z 50-2800 Linear Ion Trap mode. Mass Resolution:>3000 at m/z 609 at 250 amu/sec
4. Waters Alliance e2695 Separations with 2998 Photodiode Array (PDA) Detector

Sample Analysis

Charges & Sample requirement

Application Form

Faculty & Technical Personnel

• Dr. Sanjeev Kanojiya, Principal Scientist & Associate Professor (AcSIR), Facility-in-charge
• Ms. Pooja Soni, Technical Officer
• Mr. Tofan Kumar, Technical Officer
• Mr. Vipin Kumar, Technical Assistant
• Mr. S. A. Singh, Sr. Technician
• Mr. D. N. Vishwakarma, Sr. Technician

Research Fellows:

• 1. Mr. Yatendra (CSIR-SRF)
• 2. Ms. Priyanka Rawat (UGC-JRF)
• 3. Ms. Pratibha Singh (ICMR-SRF)
• 4. Ms. Vijaya Shukla (ICMR-SRF)
• 5. Mr.Ashish Singh (UGC-JRF)
• 6. Mr. Vikaskumar Gond (AYUSH-JRF)
• 7. Mr. Akhilesh Kumar (UGC-JRF)

Selected Publications:

1. 1. Singh, Y., et al., Structural Analysis of Diastereomeric Cardiac Glycosides and Their Genins Using Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry. Journal of the American Society for Mass Spectrometry, 2021. 32(5): p. 1205-1214.


2. 2. Jeet, A., et al., Strategies for indole alkaloids enrichment through callus culture from Alstonia scholaris (L.) R. Br. Plant Growth Regulation, 2020. 90(2): p. 383-392.


3. 3. Joshi, T., et al., Pyranocarbazoles from Murraya koenigii (L.) Spreng. as antimicrobial agents. Nat Prod Res, 2018. 32(4): p. 430-434.


4. 4. Joshi Trapti, et al., Quantitative Analysis of Bioactive Carbazole Alkaloids in Murraya koenigii. Natural Product Communications, 2015. 10(2): p. 293-295.


5. 5. Kanojiya, S. and K.P. Madhusudanan, Rapid identification of calotropagenin glycosides using high-performance liquid chromatography electrospray ionisation tandem mass spectrometry. Phytochem Anal, 2012. 23(2): p. 117-25.


6. 6. Kanojiya, S. and P.P. Yadav, Fragmentation patterns of newly isolated cassane butenolide diterpenes and differentiation of stereoisomer by tandem mass spectrometry. Journal of Mass Spectrometry, 2008. 43(10): p. 1413-1420.


7. 7. Madhusudanan, K.P., S. Kanojiya, and B. Kumar, Effect of stereochemistry on the electrospray ionization tandem mass spectra of transition metal chloride complexes of monosaccharides. Journal of Mass Spectrometry, 2005. 40(8): p. 1044-1054.