Phytochemical Finger printing

Phytochemical Finger printing

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  Phytochemical finger printing
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  INTRODUCTION  Fingerprinting methodis used to highlight the profiles of the sample matrix, which often is sufficient to provide indications of the source and method of preparation.  In herbal medicines, the profile depends not only on the preparation processes, but also on the quality of the crude herb source material.  The uniformity and stability of the chemical profiles thus represent the quality of the raw herbs. In both good agricultural practice (GAP) and good manufacturing practice (GMP).  Fingerprinting analysis is used to appraise the quality of the herbal material.
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  OBJECTIVES  The fundamentalobjective generally is to develop links between marker compound-based chromatographic or spectroscopic profiles with the efficacy of herbal products.  Thin layer chromatography (TLC) and HPTLC has been the most widely used classical method for fingerprinting analysis in herbal medicines.  In the chemical fingerprinting method, wherever possible, the bioactive compounds or important chemical marker compounds are identified to allow consistent batch-to-batch fingerprinting analysis.  GC–MS or LC–MS can be used to detect and confirm the identity of these trace marker compounds.
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  TLC and HPTLC Thin Layer Chromatography (TLC) is frequently used for the analysis of herbal medicines since various pharmacopoeias.  TLC is used as an easier method of initial screening with a semi quantitative evaluation together with other chromatographic techniques.  High performance thin layer chromatography as the advantages of many-fold possibilities of detection in analyzing herbal medicines.  The advantages of using HTLC to construct the fingerprints of herbal medicines are its simplicity, versatility, high velocity, specific sensitivity and simple sample preparation.  It summarized the progress in forced-flow-planer chromatography (FFPC) and demonstrated the importance of the different techniques like rotation planar chromatography (RPC), overpressured-layer chromatography (OPLC)
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  HPTLC Fingerprint ofGinko biloba (http://mitchj.info/suggest/camag-tlc-hptlc.html accessed 2/7/19)
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  GAS CHROMATOGRAPHY  Gaschromatography (GC) is usually used for fingerprint analysis when pharmacologically active components in herbal medicines are volatile chemical compounds.  The advantage of GC clearly lies in its high sensitivity of detection for almost all the volatile chemical compounds.  This is true for the usual FID detection and GC-MS. Furthermore, the high selectivity of capillary columns enables separation of many volatile compounds simultaneously within comparatively short times.
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  GC Fingerprint ofCinnamon Zare, R., Nadjarzadeh, A., Zarshenas, M. M., Shams, M., & Heydari, M. (2019). Efficacy of cinnamon in patients with type II diabetes mellitus: A randomized controlled clinical trial. Clinical Nutrition, 38(2), 549-556.
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  HIGH PERFORMANCE LIQUIDCHROMATOGRAPHY  HPLC can be used to analyze almost all the compounds in the herbal medicines.  Reversed- phase (RP) columns may be the most popular columns used in the analytical separation of herbal medicines.  The photodiode array detector (PDA) scans each of the ingredients eluted from the column from 200-800 nm, covering the entire range of UV visible electro magnetic radiation. This over comes the disadvantage of single wavelength analysis.  New methods like High-Speed Counter-current Chromatography (HSCCC), low- pressure size-exclusion chromatography (SEC), reversed-phase ion- pairing HPLC (RP-IPC-HPLC) provide new opportunities for good separation for some specific extracts of some herbal medicines
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  HPLC Fingerprint of Ashwagandha KThakur, A., Dey, A., S Chatterjee, S., & Kumar, V. (2015). Reverse Ayurvedic pharmacology of Ashwagandha as an adaptogenic anti-diabetic plant: a pilot study. Current Traditional Medicine, 1(1), 51-61.
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  HYPHENATED TECHNIQUES  Formost (trace-level) analytical problems in research field of traditional herbal medicines, the combination of column liquid chromatography or capillary gas chromatography with a UV-VIS or mass spectrophotometer become the preferred approach for the analysis of herbal medicines.  Various hyphenated procedures used for the analysis of herbal drugs are HPLC-DAD, CE-DAD, GC-MS, LC-MS, HPLC-MS, HPLC-DAD-MS, HPTLC-MS UHPLC−DAD−MS/MS and LC-DAD-MS.  The data obtained from such hyphenated instruments are generated two types of data and spectrum, which could provide much more information about wanted data.  Technique are more reliable authentic as compared to classic one way chromatography and spectroscopy.
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  UHPLC−DAD−MS/MS fingerprints of flowers Salminen,J. P. (2018). Two-dimensional tannin fingerprints by liquid chromatography tandem mass spectrometry offer a new dimension to plant tannin analyses and help to visualize the tannin diversity in plants. Journal of agricultural and food chemistry, 66(35), 9162-9171.
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  Fingerprinting protocol for structuralelucidation 1. A total of 25 different species of Ephedra are used. 2. Place ground plant material (~500 mg) in a 15 mL polypropylene conical tube with 6.0 mL of acetone, and sonicate for 15 min. 3. After sonication, centrifuge the sample for 10 min, and transfer the supernatant to a sample vial. 4. Repeat the extraction twice, combine the respective supernatants, and remove the solvent (acetone) by rotary evaporation. 5. Add absolute ethanol (5.0 mL) to the dried extract and let the extract dissolve.
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  6.Once the extractis dissolved, filter 2.0 mL (the first 0.5 mL should be discarded) through a 45 mm Nylon filter into an HPLC vial for analysis. 7. Carry out the HPLC–PDA analysis (the sample injection volume = 10 mL) on a Waters Alliance 2695 Separation module with a Waters 996 PDA detector, using a Waters XTerra RP18 5 mm column (4.6 mm × 150 mm) and the mobile-phase, isocratic water:ACN = 75:25 for 10min, gradient water:ACN = 75:25 to 100% ACN over 45 min, and isocratic 100% ACN for10 min, flow rate = 1 mL/min. 8. Monitor the chromatograms at three different wavelengths, 210, 254, and 320 nm, and analyze by the Waters Millenium32 software. 9. Match the retention time and the UV spectrum obtained for individual peak against known ephedrine alkaloid standards, perform quantification based on peak area of individual peaks of known alkaloids, mainly (-)-ephedrine. 10. Validate this fingerprint method by testing a number of populations within a single species of Ephedra.