Review on hyphenated techechniques.

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1. REVIEW ON HYPHENATED TECHNIQUES Bala Sunday Z BUK

2. Introduction to hyphenated techniques What is HP Tech.?? Hyphenated technique (HP Tech.) can be said to be the combination (or) coupling of more than one different analytical techniques in a single instrument with the help of proper interface and separation proficiency. Note….. In most common hyphenated techniques, mainly chromatographic techniques are combined with spectroscopic techniques (Patel et al , 2010) e.g. the GC-MS or the LC-MS

3. Introduction to hyphenated techniques The term “Hyphenated techniques” It was first adapted by Hirsch Feld in 1980 to describe a possible combination of two or more instrumental analytical methods in a single run (Hirschfeld, 1980). Why HP tech. was introduce…. The aim of the coupling is to obtain an information-rich detection for both identification and quantification compared to that with a single analytical technique (Ruchira Chin hole et al , 2012 ). spitter , sensitivity, flow-volume ratio, computer aided program

4. Advances in HP Tech…. In recent years, hyphenated techniques have received ever-increasing attention as a means to solve complex analytical problems. The power of combining separation technologies with spectroscopic techniques has been demonstrated over the years for both quantitative and qualitative analysis of unknown compounds in complex natural product extracts or fractions

5. Type of Hyphenated techniques Advances in the field of HP Tech. has exceed coupling two separation and detection techniques, in the recent time, there has been the introduction of double hybrid, also called triple hyphenated techniques

6. Composition of a HP Techs…

7. Review of Double hyphenated techniques & their Advances Gas Chromatography – Mass Spectroscopy (GC-MS ) This is a hyphenated technique developed from the coupling of GC and MS. The use of a mass spectrometer as the detector in gas chromatography was developed during the 1950s after being originated by James and Martin in 1952. (James et al , 1952)

8. GC-MS is a separation technique applicable to compounds which can be volatilized in a gas stream The advancement in GC/MS instruments using quadrupole technology had become both essential to chemical research and one of the foremost instruments used for organic analysis - identification of molecular ion. Used for confirmation of purity and identity of the components by measuring exact mass and calculating elemental composition Advances in GC-MS

9. Liquid Chromatography – Mass Spectrometry (LC-MS) Definition……. LC-MS is a chemistry technique that combines the physical separation of Liquid chromatography (or HPLC) with the Mass spectroscopy ( Sheetal et al . ). With advances in HP tecs , various types of LC-MS systems incorporating different types of interfaces are available commercially

10. Advances in L C-MS Advances in HP Tech. with different interfaces are: Designed for adequate nebulization V aporization of the liquid, I onization of the sample R emoval of the excess solvent vapor And extraction of the ions into the mass analyzer.

11. Advances in LC-MS The widely used interfaces, especially in relation to natural product analysis: Electrospray ionization ( ESI), and A tmospheric pressure chemical ionization (APCI). The latter is considered as ''the chromatographer's LC-MS interface'' because of its high solvent flow rate capability, sensitivity, response linearity, and fields of applicability. With these interfaces, various types of analyzers, e.g., quadrupole, ion trap, or TOF, can be used

12. Liquid Chromatography – Nuclear Magnetic Resonance (LC-NMR) As a result of advances in LC-NMR, this method is a powerful tool used in many areas such as natural products, organic molecules, biomolecules, drug impurities, by-products, reaction mixtures, and drug degradation products Direct coupling of liquid chromatography (LC) to NMR (Figure 5) using stop flow method was reported in 1978 (John et al, 2000)

13. Advances in LC-NMR Recent advances in both hardware and software for the direct coupling of LC and NMR have given a new life to this hyphenated technique. Advances…. These advances include: A new coil and flow cell design for high sensitivity, N ew RF system for multiple solvent suppression I mproved dynamic range gradient elution capability, Automatic peak-picking/storing capabilities. Giving structural i mformation no other techniques can!!

14. Liquid Chromatography – Infrared Spectrometry (LC-IR) LC-IR is a double hyphenated technique developed from the combination of LC and the detection method infrared spectrometry (IR) or FTIR is known as LC-IR or HPLC-IR

15. Advances in LC-IR Important For identification of organic compounds, because in the mid-IR region the structures of organic compounds have many absorption bands that are characteristic of particular functionalities, e.g., -OH, -COOH, and so on Advances in FT-IR is in the geometry of the sample during the measurement process, improving the signalto-noise ration by four times (Nagajyothi et al .). This is because the IR detector is total light limited.

16. Capillary Electrophoresis – Mass Spectrometry (CE-MS) The coupling of CZE with electrospray ionization mass spectrometry (ESI-MS), used particular for obtaining molecular mass information and structural details’

17. Advances in LC-IR Most importantly… Capillary electrochromatography (CEC) is another technique which is currently undergoing a rapid phase of advancement and development, revived by Jorgenson and Lukas in 1981 This technique has recently become more diffuse due to a number of advances in both CE instruments and detection techniques. However, on-column UV detection and in-column laser-induced Fluorescence detection remain the most commonly used methods. Despite its high sensitivity, the latter method is subjected to interferences by buffer Fluorescence ( Levsen et al , 2005).

18. Conclusion The remarkable advances in HP Tech. analytical methods over the last two decades have significantly broadened their applications in the analysis of compounds. Advances in HP Tech. such as LC-MS, GC-MS, LC-NMR, CE-MS and ICP-MS have been made to excellently solve various complex analytical problems in different fields. These techniques solve such problems in time efficient manner, h igher degree of automation, higher sample throughput, better reproducibility etc.

19. Thank you all for listening…….& Invitation