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Processing less than 100 samples Processing Large (>10ml) volume samples 25mg to 2g weight beds 1ml to 10ml volumes and Luer compatibility Ideal for a wide variety of sample types ↓ 請選用HyperSEPTM Column ↓ |
Processing more than 100 samples Processing small (<10ml) volume samples 10mg to 100mg weight beds 1ml or 2ml volume removable wells Customizable for use in a high throughput ...or method development environment ↓ 請選用MultiSEPTM 96 WELL PLATE ↓ |
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步驟2. 針對分析物選定合適之SPE column(選擇原理同HPLC Column 之選擇) |
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Reversed Phase Normal Phase Ionic Exchange |
Retain Varify (Mixed-mode C8 & SCX) Hypercarb |
步驟3. 選擇適當容量之載床(Sorbent Bed) |
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樣品的量愈大及汙染程度愈高選擇載床愈大 載床之吸附力為其重量之1% ~ 5% (i.e. 載床100mg可吸附1 ~ 5mg) 載床之大小決定樣品總萃取量，一般為2~10倍載床量，建議100mg載床處理250μl量 HyperSEP Column 可提供載床25mg ~ 2g, MultiSEP PLATE 可提供載床 10mg ~ 100mg |
步驟4. 選擇我們的Kit以方便建立你的方法SPE固相萃取方法可依以下5步驟進行: |
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1. 樣品前處理 2. Column調整 (Conditioning) 3. 注入樣品 |
4. 清洗 5. 樣品溶出 (Elution) |
為什麼要選擇Thermo Hypersil的SPE裝置?
Advantages:
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● Complete Flexibility ● Longer Column Life Times ● Powerful Contaminant Removal |
● Greater Recovery ● Better Reproducibility ● More Sensitivity |
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Low Throughput ~ HyperSEPTM COLUMNS ● Large Sample Volumes ● Lower Solvent Usage ● High Bed Capacity |
High Throughput ~ MultiSEPTM 96 WELL PLATES ● Samll Sample Volumes ● Automation Friendly ● Rapid Processing |
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INTRODUCTION TO SOLID PHASE EXTRACTION |
Solid phase extraction (SPE) improves sample purity and quantitation and can prolong the life of the analytical column. An SPE column consists of a bed of large particle sized sorbent, held between two fitted disks in a disposable tube. A variety of body shapes allow either manual extractions (using a syringe to push the sample through the adsorbent), or automated extraction using a vacuum. By choosing the correct adsorbent, the analytes of interest can be retained on the column whilst the matrix is washed out, or they can be held on the column for later elution.
SPE allows sample pre-concentration to be performed with the minimum risk of contamination or sample loss. As well as soluble contaminants, many samples also benefit from filtration prior to injection. The choice of adsorbent phase to use is governed by exactly the same principle that applies to choosing the analytical column, as SPE products are available in a similar variety of phases.
IMPORTANT CONSIDERATIONS IN METHOD DESIGN |
FLOW RATE :
Appropriate flow rates must be used with HyperSep SPE columns and wells to ensure proper bed conditioning, sample loading and elution. In general, columns can be conditioned at a flow rate up to 25mL/min. Sample loading and elution is best performed at flow rates below 10mL/min. Recovery may still be adequate at flow rates up to 20mL/min, but exceeding this may give unsatisfactory results. For ion exchange mechanisms, slower flow rates are recommended, typically 1 to 2mL/min.
SAMPLE CAPACITY :
Sorbent bed weights should suit the sample size and the separation strategy being used. Sorbents typically have a total capacity of up to 100mg for strongly retained compounds (this includes interference compounds). Less retained compounds can elute too early if the sorbent bed is too small (relative to the sample matrix volume).
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Type
of column |
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Ion
Exchange |
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Application |
Isolate non-polar compounds from a polar matrix. |
Isolate polar compounds from a non-polar matrix. |
Isolate charged (or potentially charged) compounds. |
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Absorbent Phase |
C18, C8, Retain, Hypercarb, Verify, Phenyl |
Silica, NH2, Florisil, Hypercarb |
SAC, SCX, Verify |
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Column Conditioning |
Rinse column with 3-5mi methanol followed by 3-5mi de-ionised water or buffer. |
Rinse column with 3-5mi non-polar solvents. |
Rinse column with 3-5mL de-ionised water or low ionic strength buffer. |
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Sample Loading |
Apply sample to the top of the column and draw through the bed at a suitable flow rate. |
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Column Wash |
Wash with 5 mL polar solvent. |
Wash with 5 mL non-polar solvent. |
Wash with 5mL de-ionised water or low ionic strength buffer. |
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Sample Elution |
Elute with 1-5mL non-polar solvent. |
Elute with 1-5mL polar solvent. |
Elute with 1-5mi buffer at high ionic strength (0.5-1M) or modified pH, such that the analytes are unchanged. |
ORDERING INFORMATION |
Phase：C18、C8、Silica、Phenyl、NH2、Retain (Highly cross-linked styrene-divinylbenzene co-polymer)、Florsil (powered magnesium silica gel)、SAX、SCX、Verify (C8/SO3-)、Hypercarb
Type：3 Types ~ HYPERSEP、HYPERSEP S、Multi-Sep
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