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SAMPLE PRERATION TECHNIQUES IN TRADITIONAL CHINESE MEDICINES(TCM) (HERBAL…
SAMPLE PRERATION TECHNIQUES IN TRADITIONAL CHINESE MEDICINES(TCM)
HERBAL MATRIX
ULTRASOUND-ASSISTED EXTRACTION (UAE)
extraction technique with lots of physicochemical effect
ULTRASOUND-ASSISTED ENZYMATIC EXTRACTION (UAEE)
used enzyme to break down the cell walls
high temperature that may affect on enzymatic activity
simple manipulation and less investment costs
IONIC LIQUIDS-BASED ULTRASOUND-ASSISTED EXTRACTION (IL-UAE)
as an environmentally friendly extraction technology
ionic liquid is able to accelerate the dissolution of the cellulose
high viscosity of ILs decreased the mass transfer
but it can be reduce by cavitation of ultrasound
ULTRASOUND -ASSISTED AQUEOUS TWO-PHASE EXTRACTION (UAATPE )
combined the extraction with separation
can extract bioactive compound from TCMs
two-phase system (ATPS) can be formed by 2 polymers ( polymer and salt , IL and salt , low molecular weight alcohol and salt mixed )over a limit concentration thus forming 2 immiscible phases.
MICROWAVE-ASSISTED EXTRACTION (MAE)
increase the heat
MICROWAVE-ASSISTED AQUEOUS TWO-PHASES EXTRACTION (MAATPE)
green,efficient and promising strategy for TCMs extraction
ATPS had a strong ability of absorbing microwave energy
due to much water in it , contribute rapid heating and enhanced penetration pf the extraction solvent into TCM
*applied MAATPE and ATPS consist of 21%(w/v) ethanol and 22.0% (w/v) ammonium sulafate to the extraction of polysaccharide.
IONIC LIQUIDS-BASED MICROWAVE-ASSISTED EXTRACTION (IL-MAE)
combined the unique properties of ILs an microwave heating mechanism
improved the extraction yield
shortened the extraction time
a series of green guanidinium ILs were used in IL-MAE
SOLVENT-FREE MICROWAVE EXTRACTION
combination of MAE and dry-distilation (a propose as a green method for the extraction of essential oil without using solvent)
increased intracellular pressure led to rupture the cell wall and release the compound
water-cooled oil collector as the second cooling was fix to reduce the heat loss
PRESSURIZED LIQUID EXTRACTION (PLE)
improved the solubility of the substances and the mass transfer of the solvent.
degradation of some active ingredient will occur due to high temperature during PLE
used water as an extraction solvent.( pressurized hot water, SWE or superheated water extraction)
decreasing dielectric constant, which varied the range of the polarities of extracted ingredients due to high temperature and pressure of solvent .
SUPERCRITICAL FLUID EXTRACTION (SFE)
extraction of bioactive ingredients
carbon dioxide as an extraction solvent
rapid extraction, reduce the consumption of organic solvent, easy to eliminate CO2 by reducing the pressure.
it can avoid the potential of degradation caused by long exposure to the high temperature and the atmospheric oxygen
low polarity of CO2 will lead to inefficiency of the extraction of polar compound
suitable for the extraction of the low polar and thermolabile substances
can be combined with PLE to improve the extraction efficiency
LIQUID MEMBRANE EXTRACTION (LME)
a membrane separation technology
low consumption of solvent, high enrichment and good selectivity .
enriched compound in aqueous phase can directly transfer into analytical instrument (HPLC)
SYNERGISTIC EXTRACTION TECHNIQUE
combination of 2 and more extraction technologies
had higher extraction efficiency than a single extraction technology
can overcome the shortcomings of single extraction technology and expand the application scope of each extraction technology.
BIOLOGICAL MATRIXES
MICRODIALYSIS
applied to analysis of pharmaceutical in live organism
during microdialysis, small molecular analytes could be transferred from body fluid to the dialysate through a semi-permeable membrane
advantage : continuous & real-time sampling, small damage to animals, automatic removal of proteins and lipids.
extract peurarin and the peurarin dendrimer complex from alive rabbit aqueous humor
applied to the analysis of oleanolic acid and ursolic acid in rat blood
MICROFLUIDIC TECHNIQUES
characteristic : laminar flow and liquid droplets
device : lab-on-a-chip ; integrated the sampling, dilution, reaction, separation and analysis in one step.
suitable for the SP of complex bio-fluid matrixes instead of extract active ingredient of TCMs
-a dual-channel microfluidic chip with two biomarkers(thrombin and angiotensin cenverting enzyme) ; identify bioactive phytochemical bound to enzyme from botanical drug
composed of 2 parallel channel linked by fluidic bridge
firstly, 1 channel allowed for the fomation of enzymatic complex
then, subsequent enzymatic reaction taken place at other channel(binding & separation enzymatic complex and target compound)
-last, enzymatic activities used to indicate drug quality
the bound phytochemicals dissociated from enzyme was identified using LC-MS
HERBAL AND BIOLOGICAL MATRIXES
Solid-Phase Extraction
widely accepted for cleaning up and preconcentration of TCM sample
based on selectively solid-phase adsorption
significantly reduce matrix effect during analysis
commonly used sorbents are low specificity and limited detection capability
SPE with Molecularly Imprinted Polymers
applied to extraction of active compunds form TCM with high selectivity and efficiency
show great ability of concentration in analysis of TCM in biological matrixes
efficient approach for preconcentration of analytes from 2 complex matrixes
SPE with Nanomaterials
ideal sorbents for SPE
attracted attention in pretreatment of TCM
showed good sensitivity
Matrix Solid-Phase Dispersion
widely applied to extraction of active compounds from TCM
simple and flexible technique
integrated the rupture of cells, extraction and purification contributing to less loss of samples
MSPD with Molecularly Imprinted Polymers
high selectivity and affinity
solved problems of interferences of non-target substances
applied in extraction of TCM in biological matrixes
obtain the lowest matrix effect compared to others
MSPD with Nanomaterials
enhanced the ability of absorbing target analytes in TCM of erbal matrixes
enriched low concentrations of analytes in biological matrixes
reduce the strong matrix interferences
Solid Phase Microextraction
had an ability to isolate and preconcentrate analytes of interest from TCM
facilitating the methode of sampling
low extraction capacity of those coatings limited the application of SPME in extraction for TCM
SPME with Nanomaterials
reduced matrix effect resulting from good enrichment and clean-up
enhanced the interaction between the sorbent and analytes
Headspace SPME
could effectively avoid pollution of non-volatile contents in complicated matrixes
appropriate for extraction of volatile components
Solid Phase Dynamic Extraction
commercially available inside-needle device
has high sensitivity
short sample preparation and extraction time
high sample throughput
reflecting full automation of the method
Liquid Phase Microextraction
Three Type of LPME
:
single drop microextraction (SDME)
dispersive liquid-liquid microextraction (DLLME)
hollow fibre LPME
Advantages
little consumption of organic solvents
high enrichment factors
easy contribution to the subsequently automatic analysis
a rapid and efficient extraction technique applied to the pretreatment of TCMs
ONLINE COUPLING OF SP TECHNIQUES WITH CHROMATOGRAPHY AND SPECTROMETRY
advantages : automation, high throughput analysis and time saving.
avoided the artificial error, reduce the oxidative decomposition of active compound from TCMs.
for herbal, most common chromatography coupled is counter-current chromatography(CCC)
UADE-CCC-CPC is continuous process of extraction and separation.
which has 3 procedure ;
herbal material and extraction solvent in extractor UADE
extract from first step was pumped into CCC and target compound separated
separated compund pumped into CPC for further separation
SPE-LC used to develop the chromatographic fingerprint method.
SFE-SFC method for extraction and analysis of 4 major aromatic constituent in vanilla.
A Microfluid Chip-electrophoresis-assisted biosensor strategy for simultaneous label-free multi-drug screening of G-quadruplex active ligand from 7 monomer of TCMs, 5 G-quadraplex active ligand identified
SPE-chiral LC-MS/MS for simultaneously enantioselective determination of main active pyranocoumarins
Cloud Point Extraction
based on solubilization and cloud point phenomenon of surfactants
separated hhydrophobic and hydrophilic substances through phase disengagement
applied widely to extraction of active substances in herbal matrixes
utilized for SP of TCM in biological samples
difficult to remove surfactants used which would interfere the analysis