Jagorar Yawo: Jagorar Tsohon Sojan Lab zuwa Matakan Wayar hannu ta HPLC

Gabatarwa

Na kasance a inda kuke - ina kallon mai duba a karfe 2 na safe, ina kallon wani jirgin ruwa da ya ɓace a teku, ina mamakin ko "HPLC doo-hickey" an ƙirƙira shi ne kawai don gwada hankalina. Idan kuna jin daɗin ƙoƙarin kunsa kanku a kusa da babban aikin ruwa chromatography (HPLC), ɗauki numfashi. Ba kawai akwati na sassa masu tsada ba; rawa ce mai laushi tsakanin sunadarai da physics.

A cikin dakin gwaje-gwaje, sau da yawa muna cewa lokacin wayar hannu shine "layin rayuwa" na tsarin. Ba tare da shi ba, samfurin ku yana zaune ne kawai a cikin injector, ba zai tafi ko'ina ba. Amma ta yaya za ku zaɓi "jirgin ƙasa" daidai don ɗaukar kwayoyin ku zuwa mai ganowa? Bari mu karya shi kamar muna hira a kan kofi a cikin dakin hutu.

1. Menene HPLC, Gaskiya?

A ainihin sa, HPLC shine game da rabuwa. Ka yi la'akari da shi kamar tseren inda "waƙa" shine lokaci na tsaye (shafi) kuma "iska" da ke tura masu gudu tare shine lokaci na wayar hannu na hplc.

Sihiri yana faruwa ne saboda kwayoyin halitta daban-daban suna da “alamomi” daban-daban. Wasu kwayoyin halitta suna son lokacin wayar hannu kuma suna zuƙowa cikin ginshiƙi. Wasu kuma suna da "manne" - suna da sha'awar lokaci mai tsayi kuma suna da baya. Wannan bambancin gudun shine abin da ke haifar da waɗannan kololuwa akan chromatogram ɗin ku.

2. Al'ada Phase vs. Juya Mataki

Yawancin lokaci a nan ne kowa ya fara cire gashin kansa. A cikin shekarun aikina, na sami hanya mafi sauƙi don tunawa da wannan ita ce kallon polarity.

A. Chromatography na al'ada

Wannan ita ce hanyar "tsohuwar makaranta". A cikin yanayin chromatography na al'ada, lokaci na tsaye shine iyakacin duniya - yi tunaninsa kamar maganadisu don ƙwayoyin igiya. Tsarin wayar hannu da muke amfani da shi anan ba polar bane, yawanci abubuwan kaushi kamar hexane ko heptane.

• Ma'anar: Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwararren Ƙwaƙwalwa na Ƙaƙwalwa na Ƙaƙwalwa na Ƙunƙara na Ƙunƙara na Ƙunƙara na Ƙiƙa ne na Ƙunƙara na Ƙunƙarar Ƙimar Ƙiƙwalwa na Ƙunƙarar Ƙiƙa ) yana riƙe da shi. Kwayoyin da ba na polar ba suna wankewa da sauri tare da lokacin wayar hannu mara iyaka.

• Tukwici na Pro: Idan kun haɓaka polarity na lokacin wayarku (ƙara ɗan isopropanol), samfuran ku za su yi sauri da sauri.

B. Juya lokaci Chromatography

A yau, wannan shine ma'aunin zinariya. Wataƙila kashi 90% na aikin da nake yi a cikin dakin gwaje-gwaje ya ƙunshi juzu'i na chromatography. Anan, muna jujjuya rubutun. Rukunin ba iyakacin duniya ba ne (hydrophobic, kamar mai), kuma tsarin wayar hannu shine iyakacin duniya (yawanci cakuda ruwa da methanol ko acetonitrile).

• Ma'anar: Ƙaunar ruwa (polar) kwayoyin halitta suna zuƙowa saboda suna ƙin ginshiƙin mai. Kwayoyin soyayya masu kitse (marasa iyaka) suna manne da ginshiƙi kamar manne.

• Dalilin da ya sa muke son shi: Yana da tsinkaya, mai yawa, kuma yana amfani da abubuwan kaushi na tushen ruwa waɗanda ke da sauƙin sarrafawa.

3. "Dokar Zinariya" na Ingancin Soyayya

Za ku ga "HPLC Grade" akan alamomin don dalili. Kada kayi tunanin amfani da darajar ACS ko wani abu ƙasa da haka. Me yasa? Domin waɗancan ƙazanta “kananan” a cikin ƙananan abubuwan kaushi za su taru akan ginshiƙi na tsawon lokaci, suna kashe ƙudurin ku da ƙirƙirar kololuwar fatalwa.

Koyaushe amfani da Milli-Q (18.2 MΩ.cm) ruwa mai daraja. Idan ruwan ku ya kasance "datti," tushen ku zai yi kama da dutsen dutse, kuma za ku yi kwana uku kuna ƙoƙarin gano dalilin.

4. Degassing da Tace: The Performance Killers

Air shine abokin gaba na lokaci na wayar hannu na hplc. Karkashin matsi na al'ada, iska tana narkar da abubuwan kaushi. Amma da zarar wannan kaushi ya faɗo famfo, waɗannan ƙananan kumfa za su iya shiga tarko a cikin bawul ɗin dubawa ko tantanin ganowa.

• Sakamako: Sauye-sauyen sauye-sauye, sauye-sauyen tushe, da lokutan riƙewa.

• Gyara: Yi amfani da ginanniyar degassers, ko sonication\/ vacuum tacewa idan kuna gudanar da gwaje-gwaje masu girma.

Tace Ba Zabi Ba. Ko da HPLC kaushi sa iya samun microscopic barbashi daga masana'antu tsari ko kwalban hula. Koyaushe tace ta cikin membrane 0.22µm ko 0.45µm kafin ya taɓa tafki. Yana da aiki na mintuna 5 wanda zai iya ceton ginshiƙi daga mutuwa da wuri.

5. Buffers da Gilashin Tarkon

A cikin juzu'i na chromatography, muna amfani da buffers don kiyaye pH barga don samfuran ionizable.

• Pro Tukwici: Idan kun yi amfani da buffer a yau, ku wanke tsarin da ruwa mai tsafta / gaurayawan kwayoyin kafin ku koma gida. Idan ba haka ba, waɗannan salts za su yi crystallize da shred your famfo hatimi.

• Tarkon Gilashi: Shin kun san cewa idan lokacin pH na wayar hannu ta hplc shine> 8.0, zai iya haƙiƙanin ions ƙarfe daga tafkunan gilashi? Canja zuwa bakin karfe ko kwantena filastik na musamman a cikin waɗannan lokuta.

Mu Samun Rabuwarku Cikakkiya

Samun lokacin wayar hannu daidai shine 80% na yaƙin a cikin HPLC. Idan kuna kokawa tare da tushen "shag carpet", matsa lamba, ko sakamakon da bai dace ba, kar ku ɓata wata rana kisa.

Na shafe dubban sa'o'i a benci don magance waɗannan batutuwan daidai. A koyaushe ina farin cikin raba ɗan shawarwarin "tabbatar da lab" don dawo da aikinku kan hanya.

• WhatsApp: 86 18338832256

• Imel:boonemi@aijirenvial.com
  

Nassoshi

• Snyder, L. R., Kirkland, J. J., & Dolan, J.W. (2011). Gabatarwa zuwa Chromatography Liquid Na Zamani. Bugu na 3. John Wiley & Sons. ("Gold Standard" don fahimtar hanyoyin rabuwar HPLC).

• Dolan, J.W. (2002). "Shirye-shiryen Wayar hannu." LCGC Arewacin Amurka, Jerin Shirya matsala. (Mahimman jagora mai amfani don tsaftacewa da tacewa).

• Aparicio, R. (Ed.). (2013). Manual na inganta HPLC. Elsevier. (Cikakken bayanai akan polarity mai ƙarfi da zaɓin buffer a cikin juzu'i na chromatography).

• Abubuwan da aka bayar na Agilent Technologies. "Jagorar Magance Matsalar Tsarin HPLC: Gujewa Kurakurai Na Farko Na Waya gama gari." Laburaren Albarkatun Fasaha.

• Kamfanin Waters Corporation. "Ka'idodin Chromatography: Matsayin Al'ada vs. Juya Mataki." Jerin Ilimi.

• Taron kasa da kasa kan daidaitawa (ICH). Q2(R1) Tabbatar da Hanyoyin Nazari: Rubutu da Hanya. (Misali don ƙididdige yanki mafi girma da kwanciyar hankali lokacin riƙewa).