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HPLC層析管柱所分離出分析物,若不具Chromophore 結構沒有UV/Vis波長吸光性時,將無法於HPLC - UV/Vis檢測器檢測之。Post-column Derivatization設備在於提供反應劑流路,備與層析管柱所分離出分析物合流至一組加溫反應圈,讓反應劑充分與“分析物”反應成為具Chromophore 結構而有UV/Vis吸光性衍生物,進而可以在HPLC - UV/Vis檢測器檢測之。

多數胺基酸,不具Chromophore結構沒有UV/Vis波長吸光性。當HPLC搭載一組Post-column Derivatization的設備,它提供Ninhydrine或 OPA反應劑的流路與層析管柱所分離出胺基酸合流,於加溫反應圈承受加溫反應成為具Chromophore結構UV/Vis檢測性衍生物,用以分析胺基酸 ( Amino Acid Analysis )。

Pickering公司提供 “後置反應設備儀器” 、特用層析管柱、反應試劑等化學品,預先配製妥,隨時可用分析反應試劑與移動相,用以搭配任何廠牌HPLC儀器執行胺基酸分析等相關產品:
(1)“後置反應儀器”PINNACLE PCX: Sigma Series Post-Column Derivatization Instrument
特色:
適用任何市售HPLC作衍生化胺基酸檢測
採用Syringe Pump無脈衝穩流
自動衝程清洗或程式設定清洗衝程
包含Column oven可作程式設定升、降溫
提升分離效應
完全非金屬管路
內含電腦化設定操作控制軟體
內含 Column Oven 具 Temperature Programming溫度升降程式設定功能,高效能胺基酸分析例,如下兩則圖示:

(a) 典型代表例 High – Efficiency Lithium for Physiological samples using temperature gradient.應用於生化原型胺基酸檢測。
1 Phosphoserine 16 Alanine 31 g-Aminobutyric acid
2 Taurine 17 Citrulline 32 Tryptophan
3 Phosphoethanolamine 18 a-Amino-n-butyric acid 33 Ethanolamine
4 Urea 19 Valine 34 Ammonia
5 Aspartic acid 20 Cystine 35 Hydroxylysines
6 Hydroxyproline 21 Methionine 36 Ornitine
7 Threonine 22 Allo-Isoleucine 37 Lysine
8 Serine 23 Cystathionine 38 1-Methylhistidine
9 Asparagine 24 Isoleucine 39 Histidine
10 Glutamic acid 25 Leucine 40 3-Methylhistidine
11 Glutamine 26 Tyrosine 41 Anserine
12 Sarcosine 27 Phenylalanine 42 Carnosine
13 a-Aminoadipic acid 28 b-Alanine 43 Arginine
14 Proline 29 b-Amino-i-butyric acid 44 Glucosaminic Acid*
15 Glycine 30 Homocystine 45 2-Aminoethyl-cysteine (AEC)*
(b) 典型代表例 30 – minutes High Efficiency Sodium Cation-Exchange Column for Protein and Collagen Hydrolysate samples using temperature gradient 膠原蛋白質水解成胺基酸快速檢測。
1 Aspartic Acid 11 Isoleucine
2 Threonine 12 Leucine
3 Serine 13 Norleucine
4 Glutamic Acid 14 Tyrosine
5 Proline 15 Phenylalanine
6 Glycine 16 Lysine
7 Alanine 17 Histidine
8 Cystine 18 Ammonia
9 Valine 19 Tryptophan
10 Methionine 20 Arginine
(2) “後置反應儀器”VECTOR PCX – SIGMA Series
VECTOR PCX與 PINNACLE PCX 之不同點:
採用”往復式衝程” pump, 具些微脈衝流
無 Column Oven, 須借用 HPLC 的 Column Oven 作升降溫操作 ( temperature gradient )
無電腦化設定操作控制軟體
VECTOR PCX 與 PINNACLE PCX 比較:
FEATURES PINNACLE PCX VECTOR PCX
Works with All HPLC Systems Yes Yes
All Components specifi cally designed for Post-Column Derivatization Yes No
Pump Technology Syringe Reciprocating
Piston Wash Automatic Automatic
Programmable System Flush Yes No
Column Oven Yes No
Temperature Gradient Column Oven Yes No
Automatic Column Oven Cool Down (10 min) Yes No
Electronic Valves Yes No
Check Valves No Yes
Flow Restrictors No Yes
Quick Change Reactor Cartridges Yes No
All Fluidics on Front Panel Yes No
Inert Flow Path Standard Custom
LCD Display Graphica Text
Full 32 bit PC Control Windows Software Included No
Program Storage Unlimited No
Network Enabled Yes No
Remote Diagnostics Yes Yes
Reduced Bench Space Yes No
Integrated Reagent Tray Yes No
Integrated Spill Tray Yes No
Integrated Lab Gas Manifold Yes Yes
Reduce Maintenance Costs Yes No
Reduce Reagent Usage/Costs Yes No
其他應用分析:
Pickering公司 Post-column Derivatizaion “後置反應設備儀器” 、除了應用於胺基酸分析,也成功地應用於以下藥物分析:
  Mycotoxins in feeds
  Chromium VI
Biogenic Amine   Polyether Antibiotics
Aminoglycoside Antibiotics   Guanidinos
Bromate   Hexoamines
Paralytic Shellfish Toxins   Transition / Earth Metals
Formaldehyde   Sulfa Drugs
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