CS242242B1 - Supply matrix for sample spraying into glass column packing in liquid chromatography - Google Patents
Supply matrix for sample spraying into glass column packing in liquid chromatography Download PDFInfo
- Publication number
- CS242242B1 CS242242B1 CS8539A CS3985A CS242242B1 CS 242242 B1 CS242242 B1 CS 242242B1 CS 8539 A CS8539 A CS 8539A CS 3985 A CS3985 A CS 3985A CS 242242 B1 CS242242 B1 CS 242242B1
- Authority
- CS
- Czechoslovakia
- Prior art keywords
- capillary
- axial hole
- liquid chromatography
- nut
- sample
- Prior art date
Links
- 238000004811 liquid chromatography Methods 0.000 title claims abstract description 5
- 239000011521 glass Substances 0.000 title claims description 9
- 238000012856 packing Methods 0.000 title claims description 8
- 239000011159 matrix material Substances 0.000 title description 3
- 238000005507 spraying Methods 0.000 title 1
- 238000010828 elution Methods 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 abstract description 2
- 230000006835 compression Effects 0.000 abstract 1
- 238000007906 compression Methods 0.000 abstract 1
- 230000001105 regulatory effect Effects 0.000 abstract 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 239000002594 sorbent Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N adenyl group Chemical class N1=CN=C2N=CNC2=C1N GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
Účelom riešenia je odstránenie nežiadúcich efektov sposobených postupným stlačováním a znečisťováním náplně a dosiahnutie optimálneho sposobu nanesenia vzorky na chromatograficky štipec, najma vo vysokoúčinnej kvapalinovej chromatografií. Podstata vynálezu spočívá v tom, že v prívodnej matici je vytvořený osový otvor, do ktorého je zasunutá a upevněná kapilára na přívod vzorky z dávkovacieho zariadenia, pričom vonkajší priemer kapiláry je menší ako priemer osového otvoru. Kapilára je dlhšia ako čelo matice a je vpichnutá až do náplně kolóny. Do osového otvoru je zaústěná kapilára, ktorou preteká len čistý elučný roztok cez osový otvor na čelo náplně. Poměr tokov v oboch kapilárách sa reguluje dvoma samostatnými pumpami alebo jednou pumpou s deliacim ventilom.The purpose of the solution is to eliminate unwanted ones effects caused by gradual compression and pollute the filling and reach optimum sample application method on a chromatographic pin, especially high efficiency liquid chromatography. The essence of the invention is that in the feed the nut is formed by an axial hole, into of which the capillary is inserted and fixed to feed the sample from the dispensing device, wherein the outer diameter of the capillary is smaller as the diameter of the axial hole. The capillary is longer than the face of the nut and is inserted into column charges. The axial hole is open capillary through which only pure elution flows solution through the axial hole on the filling front. ratio flow in both capillaries is regulated two separate pumps or once pump with split valve.
Description
Vynález sa týká konštrukcie prívodnej matice pre nástrek vzorky do náplně sklenenej kolony v kvapalinovej chromatografii.The invention relates to the construction of a feed matrix for injecting a sample into a glass column packing in liquid chromatography.
Jedným z rozhodujíicich faktorov určujúcich použitelnost vysokoúčinnej kvapalinovej chromatografie pre mnohonásobné sériové analýzy vzoriek je životnost chromatografie kej kolony. Je známe, že objem náplně kolony sa vplyvom vysokých tlakov, tlakových pulzov a používaných chemikálií po čase zmenšuje, pri čom sa na vstupe kolony vytvoří tzv. mrtvy objem, ktorý výrazné zhoršuje účinnost delenia. Súčasne na vstupe kolony dochádza k zachytávaniu fyzikálnych a chemických nečistot z elučného roztoku a z analyzovaných vzoriek, čím sa postupné vytvoří vrstva stlačenej a silné znečistenej náplně, ktorá znemožní ďalšie delenie. Skleněná kolona je konštrukčne riešená ako kompaktný systém, kde případné čiastočné vymenenie alebo doplnenie chýbajúcej náplně nie je možné bez poškodenia utesňovacej hlavice. Dosial známe zariadenie pre kolony tohoto typu, ktoré rieši problém znečistovania náplně, pozostáva z krátkej predkolóny predradenej před chromatografickú kolonu, pričom vzorka sa nanáša na čelo náplně. Toto zariadenie však neodstraňuje nízku účinnost delenia v dósledku vytvořeného mrtvého objemu a nedokonalý sposob nanesenia vzorky.One of the decisive factors determining the applicability of high performance liquid chromatography for multiple series analysis of samples is the lifetime of column chromatography. It is known that the volume of the packed column decreases over time due to high pressures, pressure pulses and the chemicals used. dead volume, which significantly deteriorates separation efficiency. At the same time, at the inlet of the column, physical and chemical impurities are collected from the elution solution and from the analyzed samples, gradually forming a layer of compressed and heavily contaminated packing, which prevents further separation. The glass column is designed as a compact system where any partial replacement or refilling of the missing filling is not possible without damaging the sealing head. The hitherto known device for columns of this type, which solves the problem of contamination of the packing, consists of a short precolumn upstream of the chromatography column, the sample being applied to the front of the packing. However, this device does not eliminate the low separation efficiency due to the dead volume formed and the imperfect method of sample loading.
Uvedené nedostatky odstraňuje privodná matica pre nástrek vzorky do náplně sklenenej kolony v kvapalinovej chromatografii. Podstata vynálezu spočívá v tom, že v prívodnej matici utesňujúcej kolonu je vytvořený osový otvor, do ktorého je zasunutá a upevněná kapilára na přívod vzorky z dávkovacieho zariadenia. Vonkajší priemer kapiláry je menší ako priemer osového otvoru, pričom kapilára je dlhšia ako čelo matice a je vpichnutá až do náplně kolony. Do osového otvoru je zaústěná kapilára, ktorou preteká len čistý elučný roztok cez osový otvor na čelo náplně. Vzájomný poměr tokov v oboch kapilárách sa reguluje buď dvoma samostatnými vysokotlakovými pumpami, alebo jednou pumpou s deliacim ventilom.These drawbacks are eliminated by the feed matrix for sample injection into the glass column packing in liquid chromatography. SUMMARY OF THE INVENTION An axial bore is provided in the inlet nut of the sealing column, into which a capillary for receiving a sample from the metering device is inserted and fixed. The outer diameter of the capillary is smaller than the diameter of the axial bore, the capillary being longer than the nut face and injected into the column packing. Into the axial bore is a capillary tube, through which only pure elution solution flows through the axial bore at the front of the cartridge. The flow ratio in both capillaries is controlled by either two separate high-pressure pumps or one split-valve pump.
Výhodou riešenia podl'a vynálezu je, že odstraňuje nežiadúce efekty sposobené vytvořením mrtvého objemu a svojou podstatou představuje najoptimálnejší a najúčinnejší sposob nanesenia vzorky na chromatografický stípec. S uvedeným zariadením v spojem s predkolcnou je možné chromatografickú kolonu použit pre 1 000—2 000 analýz zvlášť znečistěných vzoriek, například z biologického materiálu, namiesto bežne dosahovaných 150 — 200 analýz.The advantage of the solution according to the invention is that it eliminates the undesired effects caused by the formation of dead volume and, by its nature, represents the most optimal and efficient way of applying the sample to the chromatographic column. With this device in conjunction with the pre-lap, the chromatographic column can be used for 1,000-2,000 analyzes of particularly contaminated samples, for example of biological material, instead of the commonly achieved 150-200 analyzes.
Na pripojenom obr. 1 je znázorněný příklad zhotovenia prívodnej matice v spojení so skleněnou kolónou. Pozostáva z matice 1, ktorá sa svojim čelom 5 pritláča na teflonové tesnenie 10 sklenenej kolony 11 a je vskrutkovaná do matice 12. Do prívodnej matice 1 sú upevněné kapiláry 3, 4 pomocou skrutiek 6, 7 a těsnění 8, 9. Kapilára vzorky 3 je vpichnutá až do náplně 13 kolony 11. Elučný roztok z kapiláry 4 vytéká osovým otvorom 2 na čelo náplně 13 kolony. Skleněná kolona 11 je ukončená hlavicou 15 s vlisovanou ocelovou sieťkou 16 pomocou teflonového tesnenia 10 a je uchytená vo vonkajšom plášti 14 maticou 12 a utěsněná je prívodnou maticou 1.FIG. 1 shows an embodiment of a supply nut in conjunction with a glass column. It consists of a nut 1 which is pressed against the Teflon seal 10 of the glass column 11 with its face 5 and screwed into the nut 12. Capillaries 3, 4 are fastened to the feed nut 1 by means of screws 6, 7 and seals 8, 9. injected into the pack 13 of the column 11. The elution solution from the capillary 4 flows out through the axial opening 2 at the front of the pack 13. The glass column 11 is terminated by a head 15 with a pressed steel mesh 16 by means of a Teflon seal 10 and is held in the outer shell 14 by a nut 12 and sealed by a supply nut 1.
Vynález je možné využit najma vo vysokoúčinnej kvapalinovej chromatografii a to hlavně v prípadoch kde sa používajú elučné roztoky s vysokým obsahom solí, v oblasti pH = 6,5—7, kde dochádza k čiastočnému rozpúšťaniu silikagélu, ktorý sa najčastejšie v roznych úpravách využívá ako sorpčný materiál. Použitie je výhodné taktiež v prípadoch kde je potřebné pracovat pri vysokých tlakoch, v prípadoch keď čerpadlo vykazuje silné tlakové rázy, ale najma v prípadoch, ked sú analyzované vzorky silné znečistěné látkami, ktoré sa nevratné viažu na sorpčný materiál. Typický příklad vzoriek tohoto typu představuje biologický materiál a jeho rožne extrakty.The invention can be used especially in high performance liquid chromatography, especially in cases where high salt content elution solutions are used, in the pH range of 6.5-7, where a partial dissolution of silica gel, which is most often used as sorption sorption in various treatments material. The use is also advantageous in cases where it is necessary to operate at high pressures, in cases where the pump exhibits severe pressure surges, but in particular when the analyzed samples are heavily contaminated with substances which irreversibly bind to the sorption material. A typical example of samples of this type is biological material and its various extracts.
Nasledujúci příklad ilustruje přednosti riešenia podlá vynálezu.The following example illustrates the advantages of the present invention.
PříkladExample
Na analýzu adenínových nukleotidov z extraktu mozgového tkaniva potkana sa použila skleněná kolona 3 X 100 mm naplněná sorbentom Separon Six Cis, s priemerúm častíc 5 μώ priamo spojená s ochrannou predkolónou 3 X 20 mm naplněnou tým istým sorbentom s priemerom častíc 10 μτη. Elučný roztok obsahoval 25 mmol. I1 dihydrogenfosforečnan draselný, 4 % trietylamínu, na pH = 6,5 sa upravil s koncentrovanou kyselinou fosforečnou, nastavený prietok bol 1 mililitr . min-1, objem nastreknutej vzorky bol 20 μΐ.A 3 X 100 mm glass column packed with Separon Six Cis sorbent, with a particle diameter of 5 μώ directly coupled to a 3 X 20 mm protective pre-column packed with the same sorbent with a particle diameter of 10 μτη, was used to analyze adenine nucleotides from rat brain tissue extract. The elution solution contained 25 mmol. 1 L of potassium dihydrogen phosphate, 4% triethylamine, was adjusted to pH = 6.5 with concentrated phosphoric acid, the flow rate being 1 ml. min -1 , the injection volume was 20 μΐ.
Na obr. 2 je znázorněné rozdelenie extraktu tkaniva po niekofkých desiatkach analýz bez použitia zariadenia podfa vynálezu. Z obrázku je vidieť typické rozdvojovanie sa koncentračných zón — píkov — jednotlivých komponentov sposobené mrtvým objemom a znehodnotenou náplňou predkolóny.In FIG. 2 shows the distribution of tissue extract after several dozen analyzes without using the device according to the invention. The figure shows the typical bifurcation of concentration zones - peaks - of individual components caused by dead volume and deteriorated precolumn charge.
Na obr. 3 je znázorněný chromatogram tej istej vzorky po připojení zariadenia podfa vynálezu, pričom vzájomný poměr toku vo vstupných kapilárách bol 1: 1.· Elučný profil poslednej zóny, ktorý je podobný ideálnej gaussovskej krivke je dókazom úplného odstránenia predcházajúcich negativných javov.In FIG. 3 shows a chromatogram of the same sample after connecting the device according to the invention, the flow ratio in the inlet capillaries being 1: 1. The elution profile of the last zone, which is similar to an ideal Gaussian curve, is evidence of complete elimination of the previous negative phenomena.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS8539A CS242242B1 (en) | 1985-01-02 | 1985-01-02 | Supply matrix for sample spraying into glass column packing in liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS8539A CS242242B1 (en) | 1985-01-02 | 1985-01-02 | Supply matrix for sample spraying into glass column packing in liquid chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS3985A1 CS3985A1 (en) | 1985-08-15 |
| CS242242B1 true CS242242B1 (en) | 1986-04-17 |
Family
ID=5331888
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS8539A CS242242B1 (en) | 1985-01-02 | 1985-01-02 | Supply matrix for sample spraying into glass column packing in liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS242242B1 (en) |
-
1985
- 1985-01-02 CS CS8539A patent/CS242242B1/en unknown
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| Publication number | Publication date |
|---|---|
| CS3985A1 (en) | 1985-08-15 |
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