LC_MS.pptx

A brief description of liquid chromatography hyphenated with mass spectrometry

1. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Liquid chromatography Mass spectrometry (LC-MS) Brahmesh Reddy B R and Aishwarya G I year Ph.D Department of Plant Physiology

2. What is LC-MS? LC-MS is an analytical technique that involves physical separation of target compounds (or analytes) followed by their mass-based detection. Although relatively new, its sensitivity, selectivity and accuracy have made it a technique of choice for detecting microgram or even nanogram quantities of a variety of analytes ranging from drug metabolites, pesticides and food adulterants, to natural product extracts. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

3. LC separation LC brings about a physical separation of the analytes in a liquid sample or a solution of a solid sample. A few microliters of sample solution are injected into a flowing stream of a solvent, called the mobile phase. The mobile phase is continuously pumped through a column (a stainless-steel tube) usually filled with silica particles coated with another liquid, the stationary phase. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

4. LC separation When the sample solution-mobile phase mix reaches the column, its components will differentially interact with the stationary phase (which remains in the column) depending upon their chemical composition or physical properties. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

5. Based on the mechanism of interaction between the analyte and the stationary phase, LC separations have been classified into different modes, such as: Partition chromatography – based on the differing solubility and hydrophobicity of the analytes in the stationary phase as compared to the mobile phase. Ion-exchange chromatography – separates the analytes on the basis of their ionic charges. Size-exclusion chromatography – exploits the differences in the sizes of the analyte molecules to separate them. Affinity chromatography – separates the analytes based on their ability to bond with the stationary phase. LC separation Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

6. Liquid Chromatography A multi-component mixture that is soluble in the liquid mobile phase is separated due to the individual components’ unique partitioning between the mobile phase (Figure 1 (1)) and the stationary phase (column) (Figure 1 (3)). Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

7. Figure 1 : A simplified diagram of a liquid chromatograph hyphenated to a mass spectrometer (LC-MS) showing: (1) binary pump for mobile phase, (2) autosampler 6-port valve and injector loop, (3) column heater with column, (4) mass spectrometer detector, (5) PC Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

8. The mobile phase, typically a solvent, is used to transport the sample through the system with the aid of a high-pressure pump (Figure 1 (1)). However, it also plays a critical role in the separation process. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

9. A small volume of sample (1-100 µL) is loaded into a sample loop (Figure 1 (2)), and is then injected into the mobile phase flow by means of a six-port valve and this triggers the start of the chromatographic run. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

10. Once the sample has been injected, the mobile phase is pumped through to the column (Figure 1 (3)). A variety of column lengths (30 to 250 mm) and internal diameters (1 to 4.6 mm) are available, packed with stationary phase adsorbent materials of differing activities and particle sizes (1.5 to 10-micron diameter) that together define the column efficiency and selectivity. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

11. The column is located in a column oven; at higher temperatures (45 ºC) the viscosity of the mobile phase decreases which increases its linear velocity. This in turn reduces the run time and also improves the chromatographic resolution. Components in the mixture that have a higher affinity to the mobile phase will migrate through the column quickly with little interaction with the stationary phase. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

12. As the band of the component leaves or elutes from the column, the detector (Figure 1 (4)), will give a response that is proportional to the concentration of the component. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

13. The data acquisition system (Figure 1 (5)) records the detector response as a function of retention time in a chromatogram. The time taken between injection and detection is known as the retention time. The retention time for a component will be very specific for a given set of chromatographic conditions and may be compared with that of a standard for identification. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

14. Chromatogram Figure 2 : Chromatogram output from an HPLC or LC-MS Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

15. Chromatogram Figure 2 : Chromatogram output from an HPLC or LC-MS The peaks recorded in the chromatogram (Figure 2) are usually integrated to determine the peak area P eak area is proportional to the concentration of the component present in the sample. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

16. The mobile phase flowing out of the column (the eluent) passes through a detector that “responds” to a certain physical or chemical property, such as refractive index or light absorption, of the analytes within it. This response is captured as a signal or a “peak” whose intensity (peak area or peak height) corresponds to the amount of the component present in the sample. Chromatogram Figure 2 : Chromatogram output from an HPLC or LC-MS The time at which the detector “sees” the analyte is its RT. The identity of a compound in a sample can be confirmed by comparing its RT with the RT of a known compound. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

17. Modes of operation Isocratic Gradient An isocratic method will use the same mobile phase composition for the duration of the chromatographic run with no change in selectivity. A gradient method will enable the mobile phase composition to be changed as a function of time, which is usually optimized to either increase the chromatographic resolution or shorten run times. Considering the mobile phase, there are two main modes of operation to choose from when running a liquid chromatograph, namely, Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

18. The hyphenation of mass spectrometry to liquid chromatography Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

19. LS is best hyphenated to MS High sensitivity Linear dynamic range Selectivity High specificity Mass spectrometry is arguably the best detector that can be hyphenated to a liquid chromatograph due to its high sensitivity, linear dynamic range, selectivity and even specificity when using instrumentation with a very high mass resolving power. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

20. Mass Spectrometry Wilhelm Wien, J.J. Thomson and Francis Aston Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

21. Mass Spectrometry Figure 1: Outline of the main steps of MS and common variants available at each step Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

22. There are many different types of mass spectrometers, but they all have three features in common (Figure 1). The first is some means by which atoms or molecules from the sample can be ionized. Neutral species cannot be steered by electric fields used in mass spectrometers, and thus it is necessary to produce ions. There are many different means by which this can be accomplished, and they are collectively referred to as ion sources. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

23. There are many different types of mass spectrometers, but they all have three features in common (Figure 1). The second component of all mass spectrometers is the mass analyzer itself. There are several different means by which the m/z ratio of ions can be measured. Time-of-flight (ToF), magnetic sector and quadrupole mass analyzers are the most common, each with its own set of strengths and limitations. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

24. Demo of m/z ratio of 2,3 - dichloro toluene m/z Ratio Mass to charge ratio m / z = Mass number Charge number Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

25. There are many different types of mass spectrometers, but they all have three features in common (Figure 1). The final component common to all mass spectrometer systems is a means of detecting or counting the number of ions of a specific m/z value. These devices are called detectors and they too come in several different forms with the most common being electron multipliers, Faraday cups, channel trons and channel plates. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

26. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

27. Ion sources Component 1 Gas phase methods Electron Ionization (EI) Chemical ionization (CI) Direct Analysis in Real Time (DART) Inductively coupled plasma Desorption methods Matrix assisted Laser DI (MALDI) Fast Atom Bombardment (FAB) Thermal Ionization Sources Plasma Ionization Sources Liquid Metal Ion Sources (LIMS) Spray methods Electrospray Ionization (ESI) Desorption Electrospray Ionization (DESI) Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

28. Electrospray Ionization (ESI) Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

29. Electrospray Ionization (ESI) The sample is delivered into a capillary held at high voltage (a few kV). Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

30. Electrospray Ionization (ESI) This produces a mist of charged droplets of the same polarity. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

31. Electrospray Ionization (ESI) By using a drying gas or elevated temperatures, the charged droplets move through the source and are gradually reduced in size through evaporation of the solvent, leading to an increased surface charge density. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

32. Electrospray Ionization (ESI) At a certain point, the electric field strength within the droplet will be large enough for ions at the surface of the droplet to eject into the gaseous phase Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

33. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

34. Mass Analyzers Component 2 Time of flight (ToF) - time required Quadrupole - trajectory deflection Magnetic sensor - dispersion lll prism Ion trap - quadrupole with ringed electrodes Orbitrap - opposite cups and imagery Tandem MS - hybrid Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

35. Quadrupole mass analyzers Quadrupole mass analyzers consist of two pairs of metal rods equidistant from each other and biased at equal and opposite potentials. These twin potentials contain a fixed direct current (DC) and alternating radio frequency (RF) component, where the strength of the RF component can be varied. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

36. Quadrupole mass analyzers Any ion entering the quadrupole will have its trajectory deflected by the potential in a manner that is proportional to its m/z value. At specific RF values, only one specific m/z value will resonate with the field and be able to navigate to the end of the quadrupole and be detected. Ions with other m/z values will collide with the quadrupoles, lose their charge and not be detected. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

37. Mass Spectrometry Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

38. Ion Detectors Component 3 Electron multiplier -> x 10 8 Faraday cup (FC) -> potential drop amplified Photomultipler conversion dynode Array detectors - hybrid Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

39. Electron multipliers (EM) The essence of an EM is a serial connection of discrete metal plates called dynodes that amplifies a current of ions by a factor of ~10 8 into a measurable current of electrons. When a single secondary ion enters the EM, it is stopped by the first conversion dynode. The energy of impact is dissipated in part by ejection of electrons from the dynode material, creating an electrical charge. Additional electrons are ejected by a cascade process through subsequent dynodes. At the final dynode the accumulated charge is measured as a voltage pulse. Schematic diagram illustrating how a detected incoming ion is converted into a measurable signal using an EM detector Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

40. Faraday cup (FC) It consists of a hollow conducting electrode connected to ground through a high resistance. The ions hitting the collector cause a flow of electrons from ground through the resistor and the resulting potential drop across the resistor is amplified. The elementary charge on a single ion is 1.6 x 10 -19 C. Therefore, a count rate of 1 x 10 6 c/s (about the upper realistic limit for EM detector usage) would produce a current of 1.6 x 10 -13 A. Schematic diagram of a Faraday cup ion detector Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

41. Photomultiplier conversion dynode The ions initially strike a dynode which results in electron emission. The electrons produced then strike a phosphor screen which in turn releases photons. The photons then pass into the multiplier where amplification occurs in a cascade fashion – much like with the electron multiplier Schematic diagram of a photomultiplier conversion dynode detector Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

42. Array detectors Schematic diagram of double focusing magnetic sector mass spectrometer incorporating a multicollector system and static magnetic field – the nanoscale secondary ion mass spectrometer (NanoSIMS). Array detectors can cover a broad range of detector types and systems but can be generally broken down into two categories: detectors that can measure many ions of differing mass-to-charge ratio (m/z) values simultaneously detectors that are position sensitive Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

43. LC - MS IN A NUTSHELL Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

44. Using MS for LC detection Although a wide variety of detectors of differing technologies and sensitivities have been coupled with LC for analyzing different sample types, the mass spectrometer has emerged as a selective, sensitive and universal detector. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

45. Using MS for LC detection Unlike other detectors, the LC eluent carrying the separated analytes is not allowed to flow into the mass spectrometer. While the LC system is operated at ambient pressures, the mass spectrometer is operated under vacuum and the two are coupled through an interface. As the column eluent flows into the interface, the solvent is evaporated by applying heat and the analyte molecules are vaporized and ionized. This is a crucial step as the mass spectrometer is only capable of detecting and measuring the gas phase ions. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

46. Using MS for LC detection As the analyte ions are generated at atmospheric pressure in the interface, the process is called atmospheric pressure ionization (API) and the interface is known as the API source. Electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) are the most commonly used sources in LC-MS analysis. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

47. The analyte ions are drawn into the mass spectrometer where they are subjected to electric fields and/or magnetic fields. The flight paths of the ions are altered by varying the applied fields which ensures their separation from one another on the basis of their mass-to-charge (m/z) values. Post-separation, the ions can be collected and detected by a variety of mass detectors,2 of which the most common one is the electron-multiplier. When the separated ions strike the surface of the electron-multiplier (a dynode), secondary electrons are released. Using MS for LC detection Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

48. These secondary electrons are multiplied by cascading them through a series of dynodes. The amplified current generated by the flow of the secondary electrons is measured and correlated to the ion concentrations in the mass spectrometer at any given instant in time Using MS for LC detection Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

49. Plotting LC-MS data Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

50. How do you read an LC-MS mass spectrum and what does it tell you? Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

51. Plotting LC-MS data The abundances of the ions measured during the analysis of a sample by LC-MS are plotted as a total ion chromatogram (TIC). This plot displays the peak intensities of the analyte ions versus their RT. Further, each point in the chromatogram is associated with a mass spectrum. The mass spectrum depicts the ion abundances versus the measured m/z values Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

52. How do you read an LC-MS mass spectrum and what does it tell you? The mass spectrum of a compound not only provides information about the mass of the parent compound (from the m/z value of its ion), but also helps to elucidate the structure of the compound from the relative abundances of isotopic mass peaks. The area of the analyte peak is used for its quantification. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

53. How do you read an LC-MS mass spectrum and what does it tell you? The mass spectrometer can be operated in two modes, a) scan b) selected ion monitoring (SIM). In the scan mode, it is set to detect all the ions from low m/z to high m/z values within a specified time period. This mode is used when analyzing unknown samples or when there is no available information about the ions present in a sample. When operating in SIM mode, the mass spectrometer is set to measure specific m/z values. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

54. How do you read an LC-MS mass spectrum and what does it tell you? An LC-MS system may be run in either positive ion mode for basic analytes generating protonated molecules [M+H]+, or negative ion mode for acidic analytes generating deprotonated molecules [M-H]-. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

55. LC-MS analysis Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

56. LC-MS analysis quantification of genotoxic impurities in active pharmaceutical ingredients detection of doping agents , such as anabolic agents and simulants, in exhaled breath quantification of drug metabolites in biological fluids detection of adulterants in food materials and dietary supplements determination of alkylphenol ethoxylates (APEOs) in tannery sediments quantification of personal care products in swimming pool and river water samples quantification of nucleotides and their derivatives in bacterial cells quantification of the proteome as a rapid assay for the detection of SARS-CoV-2 Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

57. Case Study 1 Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

58. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

59. Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

60. Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

61. Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

62. Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

63. Case Study 2 Schuster O, Zvi A, Rosen O, Achdout H, Ben-Shmuel A, Shifman O, Yitzhaki S, Laskar O, Feldberg L. Specific and Rapid SARS-CoV-2 Identification Based on LC-MS/MS Analysis. ACS Omega. 2021 Jan 26;6(5):3525-3534. doi: 10.1021/acsomega.0c04691. PMID: 33585737; PMCID: PMC7857140. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

64. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

65. Mosaad I. Morsy, Eman G. Nouman, Youmna M. Abdallah, Mourd A. Zainelabdeen, Mohamed M. Darwish, Ahmed Y. Hassan, Amira S. Gouda, Mamdouh R. Rezk, Ahmed M. Abdel-Megied, Hoda M. Marzouk, A novel LC-MS/MS method for determination of the potential antiviral candidate favipiravir for the emergency treatment of SARS-CoV-2 virus in human plasma: Application to a bioequivalence study in Egyptian human volunteers, Journal of Pharmaceutical and Biomedical Analysis, Volume 199, 2021,114057,ISSN 0731-7085,https://doi.org/10.1016/j.jpba.2021.114057. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

66. Plants and LC-MS Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

67. Wang, Guodong & Wang, G.-D. (2014). Applications of LC-MS in Plant Metabolomics. 10.1007/978-94-017-9291-2_9. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

68. Jebaseelan, S. & Jose, B. & Meera, Dr.R.. (2021). Phytochemical Investigation Using LC-MS Analysis and Antimicrobial Activities of Leaf Extract of Muntingia calabura Linn. International Journal of Pharmaceutical Sciences Review and Research. 69. 10.47583/ijpsrr.2021.v69i02.006. Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry

69. Thank you Department of Plant Biotechnology MBB 504 (2+1) / Techniques in Molecular Biology - I Brahmesh Reddy B R Aishwarya G Liquid Chromatography Mass spectrometry