natgillin/translations-raw huggingface dataset:
HPLC equipment
Grelnik kolone, ki lahko temperaturo ohranja na 30 °C ± 1 °C
Gradient pumping system, capable of operating at 1,0 ml/min at 200 bar
Fluorescenčni detektor z valovno dolžino vzbujanja 330 nm in valovno dolžino emisije 440 nm
Autosampler with derivatisation capability
Integrator ali programska oprema za obdelavo podatkov, ki lahko meri površino vrha
Column heater, capable of maintaining the column at 30 °C ± 1 °C
Kolona LiChrospher ® – 100 (250 × 4,6 mm) ali enakovredna kolona, polnjena z oktadecilsilanom (C 18), velikost delcev 5 μm
Fluorescence detector, capable of operating at 330 nm excitation wavelength and 440 nm emission wavelength
VZORČENJE
Integrator or data processing software capable of peak area measurement
Vzorčenje mora biti izvedeno v skladu s standardom ISO 707.
A LiChrospher® — 100 column (250 × 4,6 mm) or an equivalent column packed with octadecylsilane (C 18), 5 μm particle size.
POSTOPEK
SAMPLING
Priprava raztopine internega standarda
Sampling shall be carried out in accordance with ISO Standard 707.
Natehtajte 30,0 ± 0,1 mg triptamin-monohidroklorida (4.2.3) v 100-mililitrsko merilno bučko (5.6) in dopolnite do oznake z metanolom (4.2.1).
PROCEDURE
Odpipetirajte 1 ml (5.3) te raztopine v 10-mililitrsko merilno bučko (5.6) in dopolnite do oznake z metanolom (4.2.1), da dobite raztopino s koncentracijo 0,15 mM triptamina.
Preparation of the internal standard solution
Priprava raztopine preskusnega vzorca
Weigh 30,0 ± 0,1 mg of tryptamine-monohydrochloride (4.2.3) into a 100 ml volumetric flask (5.6) and make up to the mark with methanol (4.2.1)
Odtehtajte 1,000 ± 0,001 g vzorca posnetega mleka v prahu v 25-mililitrsko čašo (5.2).
Pipette 1 ml (5.3) of this solution into a 10 ml volumetric flask (5.6) and make up to the mark with methanol (4.2.1) in order to obtain a 0,15 mM tryptamine concentration
S pipeto (5.3) dodajte 10 ml destilirane vode pri 40 °C ± 1 °C in mešajte 30 minut z magnetnim mešalnikom (5.4), da raztopite morebitne grudice.
Preparation of the test sample solution
Odpipetirajte 0,2 ml (5.5) rekonstruiranega vzorca mleka v 10-mililitrsko merilno bučko (5.6), dodajte 100 μl 0,15 mM raztopine triptamina (7.1) z brizgalko (5.7) in dopolnite do oznake z metanolom (4.2.1).
Weigh 1,000 ± 0,001 g of the SMP sample into a 25 ml beaker (5.2).
Previdno zmešajte z obračanjem bučke in postavite v ultrazvočno kopel (5.8) za 15 min.
Add 10 ml of distilled water at 40 °C ± 1 °C by a pipette (5.3) and stir with a magnetic stirrer (5.4) for 30 minutes in order to dissolve any lumps
Centrifugirajte (5.9) 10 minut pri 27000 × g in zberite bistro tekočino v stekleno vialo (5.10).
Pipette 0,2 ml (5.5) of the reconstituted milk into a 10 ml volumetric flask (5.6), add 100 μl of the 0,15 mM tryptamine solution (7.1) using a syringe (5.7) and make up to the volume with methanol (4.2.1).
Opomba: Raztopino preskusnega vzorca je treba do HPLC-analize hraniti pri 4 °C.
Mix carefully by inversion and sonicate (5.8) for 15 min
Priprava raztopine eksternega standarda
Centrifuge (5.9) at 27000 g × g for 10 minutes and collect the supernatant in a glass vial (5.10)
Odtehtajte 55,4 mg PEDP (4.1) v 50-mililitrsko merilno bučko (5.6) in dodajte z merilnim valjem (5.11) približno 25 ml kloroforma (4.2.2).
Note: Test sample solution should be stored at 4 °C until the HPLC analysis is performed.
Zaprto bučko segrejte do 50 °C ± 1 °C in previdno premešajte, da se PEDP raztopi.
Preparation of the external standard solution
Ohladite bučko na 20 °C in dopolnite do oznake z metanolom (4.2.1) ter zmešajte z obračanjem.
Weigh 55,4 mg PEDP (4.1) into a 50 ml volumetric flask (5.6) and add about 25 ml of chloroform (4.2.2) using a graduated cylinder (5.11).
Odpipetirajte 1 ml (5.3) te raztopine v 100-mililitrsko merilno bučko (5.6) in dopolnite do oznake z metanolom (4.2.1).
Heat the stoppered flask to 50 °C ± 1 °C and mix carefully till the PEDP dissolves.
Odpipetirajte 1 ml (5.3) te raztopine v 10-mililitrsko merilno bučko (5.6), dodajte 100 μl (5.7) 0,15 mM raztopine triptamina (7.1) in dopolnite do oznake z metanolom (4.2.1).
Cool the flask to 20 °C, make up to the volume with methanol (4.2.1) and mix by inversion
Zmešajte z obračanjem bučke.
Pipette 1 ml (5.3) of this solution into a 100 ml volumetric flask (5.6) and make up to the volume with methanol (4.2.1).
Opomba: Raztopino referenčnega vzorca je treba do HPLC-analize hraniti pri 4 °C.
Pipette 1 ml (5.3) of this solution into a 10 ml volumetric flask (5.6), add 100 μl (5.7) of 0,15 mM tryptamine solution (7.1) and make up to the volume with methanol (4.2.1).
Priprava reagenta za derivatizacijo
Mix by inversion
Odtehtajte 25,0 ± 0,1 mg OPA (4.3.4) v 10-mililitrsko merilno bučko (5.6), dodajte 0,5 ml (5.5) metanola (4.2.1) in previdno premešajte, da se OPA raztopi.
Note: Reference sample solution should be stored at 4 °C until the HPLC analysis is performed.
Dopolnite do oznake z raztopino borove kisline (4.3.2) in dodajte z brizgalko (5.7) 20 μl 2-merkaptoetanola (4.3.3).
Preparation of the derivatising reagent
Opomba: Reagent za derivatizacijo je treba hraniti v temni viali pri 4 °C in je stabilen en teden.
Weigh 25,0 ± 0,1 mg of OPA (4.3.4) into a 10 ml volumetric flask (5.6), add 0,5 ml (5.5) of methanol (4.2.1) and mix carefully to dissolve the OPA.
HPLC-določitev
Make up to the mark with boric acid solution (4.3.2) and add 20 μl of 2-mercaptoethanol (4.3.3) by syringe (5.7).
Topila (eluenti) (4.4)
Note: The derivatising reagent should be stored at 4 °C in a brown glass vial and is stable for one week.
Topilo A: 0,3 mM raztopina natrijevega dihidrogen fosfata in 3 mM natrijevega acetata (pH uravnan na 6,5 ± 0,1 z ocetno kislino): metanol: tetrahidrofuran = 558:440:2 (v/v/v)
Determination by HPLC
Topilo B: metanol
Elution solvents (4.4)
Predlagani elucijski gradient
Solvent A: Solution of 0,3 mM sodium dihydrogen phosphate and 3 mM sodium acetate solution (adjusted to pH 6,5 ± 0,1 with acetic acid): methanol: tetrahydrofuran = 558:440:2 (v/v/v)
Čas (min)
Solvent B: methanol
Topilo A (%)
Suggested eluting gradient:
Topilo B (%)
Time (min)
Pretok (ml/min)
Solvent A (%)
Začetek
Solvent B (%)
0Opomba: Elucijski gradient je morda treba rahlo spremeniti, da bi dobili ločljivost, kakor je prikazana na sliki 1.
Flow rate (ml/min)
Temperatura kolone: 30 °C.
Initial
Prostornina vbrizganja: 50 μl reagenta za derivatizacijo in 50 μl raztopine vzorca
0Note: The eluting gradient may require slight modification in order to achieve the resolution shown in figure 1.
Vzpostavitev ravnotežja v koloni
Column temperature: 30 °C.
Če se sistem začne dnevno, se kolona spira 15 minut s 100-odstotnim topilom B, potem nastavi A:B = 40:60 in 15 minut vzpostavlja ravnotežje pri pretoku 1 ml/min.
Injection volume: 50 μl derivatising reagent and 50 μl sample solution
Slepa določitev se izvede z vbrizganjem metanola (4.2.1).
Column equilibration
Opomba: Pred daljšim hranjenjem je treba kolono spirati 30 minut z mešanico metanol: kloroform = 80:20 (v/v).
Starting up the system on a daily basis, flush the column with 100 % solvent B for 15 minutes, then set at A:B = 40:60 and equilibrate at 1 ml/min for 15 minutes.
Določitev vsebnosti PS + PE v preskusnem vzorcu
Perform a blank run by injecting methanol (4.2.1).
Izvedite zaporedje kromatografskih analiz s konstantnim časom med dvema analizama, da s tem zagotovite konstanten retenzijski čas.
Note: Before long-term storage flush the column with methanol: chloroform = 80:20 (v/v) for 30 minutes.
Vsakih 5–10 preskusnih vzorcev vbrizgajte raztopino eksternega standarda (7.3) za ovrednotenje odzivnega faktorja
Determine the PS + PE content in the test sample
Opomba: Na vsakih 20–25 analiz se kolona očisti, tako da se najmanj 30 minut spira s 100-odstotnim topilom B (7.5.1).
Perform the sequence of the chromatographic analyses keeping constant the run-to-run time in order to obtain constant retention times.
Način integracije
Inject the external standard solution (7.3) every 5-10 test sample solutions in order to calculate the response factor
Vrh PEDP
Note: The column shall be cleaned by flushing with 100 % solvent B (7.5.1) for at least 30 minutes every 20-25 runs.
Vrh PEDP se pojavi samostojno.
Integration mode
Njegova površina se določi z integracijo dolina–dolina.
PEDP peak
Vrh triptamina
PEDP is eluted as a single peak.
Vrh triptamina se pojavi samostojno (slika 1).
Determine the peak area by valley-to- valley integration.
Njegova površina se določi z integracijo dolina–dolina.
Tryptamine peak
Skupine vrhov PS in PE
Tryptamine is eluted as a single peak (Figure 1).
Pri opisanih pogojih (slika 1) se PS eluira v obliki dveh delno neločenih vrhov, pred katerima je še en manjši vrh.
Determine the peak area by valley-to-valley integration.
PE se eluira v obliki treh glavnih delno neločenih vrhov.
PS and PE peaks groups
Površina vsake skupine vrhov se določi z nastavitvijo bazne linije, kakor je prikazano na sliki 1.
Under the described conditions (Figure 1), PS elutes as two main partially unresolved peaks preceded by a minor peak.
IZRAČUN IN IRAŽANJE REZULTATOV
PE elutes as three main partially unresolved peaks.
Vsebnost PS in PE v preskusnem vzorcu se izračuna, kakor sledi:
Determine the whole area of each peak cluster setting the baseline as reported in Figure 1.
kjer je:
CALCULATION AND EXPRESSION OF RESULTS
C = vsebnost PS ali PE (v mg/100 g prahu) v preskusnem vzorcu
PS and PE content in the test sample shall be calculated as follows:
A 1 = površina vrha PEDP raztopine standardnega vzorca (7.3)
where:
A 2 = površina vrha PS ali PE raztopine preskusnega vzorca (7.2)
C = PS or PE content (mg/100 g powder) in the test sample
T 1 = površina vrha triptamina v raztopini standardnega vzorca (7.3)
A 1 = PEDP peak area of the standard sample solution (7.3)
T 2 = površina vrha triptamina v raztopini preskusnega vzorca (7.2)
A 2 = PS or PE peak area of the test sample solution (7.2)
TOČNOST METODE
T 1 = Tryptamine peak area of the standard sample solution (7.3)
Opomba: Vrednosti za ponovljivost so bile izračunane v skladu z mednarodnim standardom IDF (*).
T 2 = Tryptamine peak area of the test sample solution (7.2).
Ponovljivost
ACCURACY OF THE METHOD
Relativni standardni odmik ponovljivosti, izražajoč variabilnost neodvisnih analitskih rezultatov, ki jih dobi isti analitik z isto opremo pri enakih pogojih na istem preskusnem vzorcu v kratkem časovnem intervalu, ne sme presegati vrednosti 2 % relativno.
Note: The values for repeatability were calculated according to the IDF International Standard (*).
Če sta dve določitvi pridobljeni pri teh pogojih, relativna razlika med dvema rezultatoma ne sme biti večja od 6 % aritmetične sredine teh rezultatov.
Repeatability
Obnovljivost
If two determinations are obtained under these conditions, the relative difference between the two results should not be greater than 6 % of the arithmetic mean of the results.
VIRI
Reproducibility
Slika 1
If two determinations are obtained by operators in different laboratories using different apparatus under different conditions for the analysis on the same test sample, the relative difference between the two results should not be greater than 11 % of the arithmetic mean of the results.
HPLC vzorec derivatov o-ftaldialdehida (OPA), fosfatidilserina (PS) in fosfatidiletanolamina (PE) v ekstraktu metanola rekonstruiranega posnetega mleka v prahu.
REFERENCES
Prikazan je način integracije vrhov PS, PE in triptamina (interni standard)
Resmini P., Pellegrino L., Hogenboom J.A., Sadini V., Rampilli M., ‘Detection of buttermilk solids in skimmilk powder by HPLC quantification of aminophospholipids’.
Fluorescenca (poljubne enote)
Figure 1
Dodatek II
HPLC pattern of OPA-derivatives of phosphatidylserine (PS) and phosphatidylethanolamine (PE) in methanol extract of reconstituted skim-milk powder.
ZAZNAVANJE SLADKE SIROTKE V POSNETEM MLEKU V PRAHU ZA JAVNO SKLADIŠČENJE Z DOLOČITVIJO MAKROPEPTIDOV KAZEINA S TEKOČINSKO KROMATOGRAFIJO VISOKE ZMOGLJIVOSTI (HPLC)
Integration mode for the peaks of PS, PE and tryptamine (internal standard) is reported
NAMEN IN PODROČJE UPORABE
Fluorescence (arbitrary units)
Ta metoda omogoča zaznavanje sladke sirotke v posnetem mleku v prahu, ki je namenjeno javnemu skladiščenju, z ugotavljanjem makropeptidov kazeina.
Appendix II
SKLIC
DETECTION OF RENNET WHEY IN SKIMMED MILK POWDER FOR PUBLIC STORAGE BY DETERMINATION OF CASEINOMACROPEPTIDES HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
Mednarodni standard ISO 707 – Mleko in mlečni proizvodi – Smernice o vzorčenju
SCOPE AND FIELD OF APPLICATION
Vsebnost trdnih snovi sladke sirotke je opredeljena kot masni odstotek, ki se določi z vsebnostjo makropeptidov kazeina v skladu z opisanim postopkom.
This method allows detection of rennet whey in skimmed milk powder intended for public storage by determination of the caseinomacropeptides.
PRINCIP
REFERENCE
Rekonstitucija posnetega mleka v prahu, odstranitev maščob in beljakovin s trikloroocetno kislino, ki ji sledi centrifugacija ali filtracija.
International Standard ISO 707 - Milk and Milk Products - Guidance on sampling.
Določitev količine makropeptidov kazeina (CMP) v supernatantu se izvede s tekočinsko kromatografijo visoke ločljivosti (HPLC).
The content of rennet whey solids is defined as the percentage by mass as determined by the caseinomacropeptide content by the procedure described.
Ocenjevanje rezultatov, dobljenih za vzorce, glede na standardne vzorce posnetega mleka v prahu z dodatkom znanega odstotka sirotke v prahu ali brez njega.
PRINCIPLE
Uporabljena voda je destilirana voda ali voda vsaj enakovredne čistosti.
Reconstitution of the skimmed milk powder, removal of fat and proteins with trichloroacetic acid, followed by centrifugation or filtration;
Raztopina trikloroocetne kisline
Determination of the quantity of caseinomacropeptides (CMP) in the supernatant by high-performance liquid chromatography (HPLC);
Raztopite 240 g trikloroocetne kisline (CCl3COOH) v vodi in dopolnite do prostornine 1000 ml.
Evaluation of the result obtained for the samples by reference to standard samples consisting of skimmed milk powder with or without the addition of a known percentage of whey powder.
Raztopina mora biti bistra in brezbarvna.
The water used shall be distilled water or water of at least equivalent purity.
Raztopina eluenta, pH 6,0
Trichloroacetic acid solution
Raztopite 1,74 g dikalijevega hidrogen fosfata (K2HPO4), 12,37 g kalijevega dihidrogen fosfata (KH2PO4) in 21,41 g natrijevega sulfata (Na2SO4) v približno 700 ml vode.
Dissolve 240 g of trichloroacetic acid (CCl3COOH) in water and make up to 1000 ml.
Če je potrebno, uravnajte pH na vrednost 6,0 z uporabo raztopin fosforjeve kisline ali kalijevega hidroksida.
The solution should be clear and colourless.
Dopolnite z vodo do 1000 ml in homogenizirajte.
Eluent solution, pH 6,0
Opomba: Sestava eluenta se lahko posodobi tako, da ustreza spričevalu v skladu s standardi ali priporočili proizvajalca materiala za polnitev kolone.
Dissolve 1,74 g of dipotassium hydrogen phosphate (K2HPO4), 12,37 g of potassium dihydrogen phosphate (KH2PO4) and 21,41 g of sodium sulphate (Na2SO4) in about 700 ml of water.
Raztopino eluenta pred uporabo filtrirajte skozi membranski filter s premerom por 0,45 μm.
Adjust, if necessary, to pH 6,0, using a solution of phosphoric acid or potassium hydroxide.
Raztopina za izpiranje
Make up to 1000 ml with water and homogenise.
Zmešajte eno volumsko enoto acetonitrila (CH3CN) z devetimi volumskimi enotami vode.
Note: The composition of the eluent can be updated to comply with the certificate of the standards or the recommendations of the manufacturer of the column packing material.
Pred uporabo filtrirajte zmes z membranskimi filtrom s premerom por 0,45 μm.
Filter the eluent solution, prior to use, through a membrane filter with a 0,45 μm pore diameter.
Opomba: Uporabi se lahko katera koli druga raztopina za izpiranje z baktericidnim učinkom, ki ne poslabša ločljivosti kolone.
Flushing solvent
Standardni vzorci
Mix one volume acetonitrile (CH3CN) with nine volumes water.
Posneto mleko v prahu, ki ustreza zahtevam te uredbe (npr. [0])
Filter the mixture prior to use through a membrane filter with a 0,45 μm pore diameter.
Enako posneto mleko v prahu z dodatkom 5 % (m/m) sladke sirotke v prahu s standardno sestavo (tj. [5]).
Note: Any other flushing solvent with a bactericidal effect which does not impair the columns' resolution efficiency may be used.
Analitska tehtnica
Standard samples
Neobvezna centrifuga, ki lahko razvije centrifugalno silo 2200 g, z epruvetami za centrifugiranje z zamaški ali pokrovčki s prostornino približno 50 ml
Skimmed milk powder meeting the requirements of this Regulation (i.e. [0])
Mehanični stresalnik
The same skimmed milk powder adulterated with 5 % (m/m) rennet-type whey powder of standard composition (i.e. [5])
Stekleni lijaki s premerom približno 7 cm
Analytical balance
Filtrirni papir, srednje porozen, s premerom približno 12,5 cm
Optional centrifuge capable of attaining a centrifugal force of 2200 g, fitted with stoppered or capped centrifuge tubes of about 50 ml capacity
Oprema za filtriranje iz stekla s premerom por membranskega filtra 0,45 μm
Mechanical shaker
Graduirana pipeta, ki prepušča 10 ml (ISO 648, razred A, ali ISO/R 835) ali razdelilni sistem, ki lahko prepušča 10,0 ml v dveh minutah
Glass funnels, diameter about 7 cm
Razdelilni sistem, ki lahko prepušča 20,0 ml vode pri pribl.
Filter papers, medium filtration, diameter about 12,5 cm
50 °C
Glass filtration equipment with 0,45 μm pore diameter membrane filter
Termostatska vodna kopel, nastavljena na 25 ± 0,5 °C
Graduated pipettes allowing delivery of 10 ml (ISO 648, Class A, or ISO/R 835) or a dispensing system capable of delivering 10,0 ml in two minutes
HPLC-oprema, ki jo sestavljajo:
Dispensing system capable of delivering 20,0 ml water at ca.
Črpalka
50 °C
Ročni ali avtomatski injektor s prostornino 15 do 30 μl
Thermostatic water bath, set at 25 ± 0,5 °C
Dve serijski koloni TSK 2000-SW (dolžina 30 cm, notranji premer 0,75 cm) ali istovrstni koloni (tj. enojna TSK 2000-SWxl, enojna Agilent Technologies Zorbax GF 250) in predkolona (3 cm × 0,3 cm), polnjena z I 125 ali enako učinkovitim materialom
HPLC equipment, consisting of:
Termostatiran grelnik kolone, nastavljen na 35 ± 1 °C
Pump
UV-detektor z nastavljivo valovno dolžino z možnostjo merjenja pri 205 nm z občutljivostjo 0,008 Å.
Injector, hand or automatic, with a 15 to 30 μl capacity
Integrator z integracijo dolina–dolina
Two TSK 2000-SW columns in series (length 30 cm, internal diameter 0,75 cm) or equivalent columns (e.g. single TSK 2000-SWxl, single Agilent Technologies Zorbax GF 250) and a precolumn (3 cm × 0,3 cm) packed with I 125 or material of equivalent effectiveness
Opomba: Mogoče je delo s kolonami pri sobni temperaturi, vendar je njihova moč ločljivosti nekoliko manjša.
Thermostatic column oven, set at 35 ± 1 °C
V tem primeru mora biti obseg temperature manjši od ± 5 °C v kateri koli stopnji analize.
Variable wavelength UV detector, permitting measurements at 205 nm with a sensitivity of 0,008 Å
Vzorci se vzamejo v skladu s postopkom, določenim v mednarodnem standardu ISO 707.
Integrator capable of valley-to-valley integration
Vendar lahko države članice uporabijo drugo metodo vzorčenja, če je skladna z navedenim standardom.
Note: Working with columns kept at room temperature is possible, but their power of resolution is slightly lower.
Vzorec hranite pri pogojih, ki izključujejo kakršno koli kvarjenje ali spremembo sestave.
In that case, the temperature should vary by less than ± 5 °C in any one range of analyses.
Priprava preskusnega vzorca
Samples shall be taken in accordance with the procedure laid down in International Standard ISO 707.
Prenesite mlečni prah v posodo z nepredušnim pokrovom s približno dvakrat večjo prostornino, kakor je prostornina prahu.
However, Member States may use another method of sampling provided that it complies with the principles of the abovementioned standard
Posodo takoj zaprite.
Store the sample in conditions which preclude any deterioration or change in composition
Zmešajte mlečni prah s ponavljajočim obračanjem posode.
Preparation of the test sample
Preskusni delež
Transfer the milk powder into a container with a capacity of about twice the volume of the powder, fitted with an airtight lid.
Odtehtajte 2,000 ± 0,001 g preskusnega vzorca v centrifugalno epruveto (6.2) ali ustrezno zaprto bučko (50 ml).
Close the container immediately.
Odstranitev maščob in beljakovin
Mix the milk powder well by means of repeated inversion of the container.
Preskusnemu deležu dodajte 20,0 ml tople vode (50 °C).
Test portion
Raztopite prah s petminutnim stresanjem na mehaničnem stresalniku (6.3).
Weight 2,000 ± 0,001 g of test sample into a centrifuge tube (6.2) or a suitable stoppered flask (50 ml).
Postavite epruveto v vodno kopel (6.10) in pustite, da se temperatura uravna na 25 °C.
Removal of fat and proteins
V dveh minutah dodajte 10,0 ml raztopine trikloroocetne kisline (5.1) s temperaturo približno 25 °C in pri tem intenzivno mešajte z magnetnim mešalnikom (6.4).
Add 20,0 ml of warm water (50 °C) to the test portion.
Postavite epruveto v vodno kopel (6.10) in pustite 60 minut.
25 °C in two minutes, while stirring vigorously with the aid of the magnetic stirrer (6.4).
Pred kakršno koli prekinitvijo, ki traja več kot 24 ur, spirajte kolone z vodo in nato z raztopino (5.3) pri pretoku 0,2 ml na minuto najmanj tri ure.
Place the tube in a water bath (6.10) and leave for 60 minutes
Rezultati kromatografske analize preskusnega vzorca [E] so dobljeni v obliki kromatogramov, pri katerih je vsak vrh opredeljen z retenzijskim časom RT, kakor sledi:
Centrifuge (6.2) for 10 minutes at 2200 g, or filter through paper (6.6), discarding the first 5 ml of filtrate
Vrh II:
Chromatographic determination
Drugi vrh kromatograma z retenzijskim časom približno 12,5 minute.
Inject 15 to 30 μl of accurately measured supernatant or filtrate (8.3.3) into the HPLC apparatus (6.11) operating at a flow rate of 1,0 ml of eluent solution (5.2) per minute
Vrh III:
Note 1.
Tretji vrh kromatograma, ki ustreza CMP z retenzijskim časom približno 15,5 minute.
Another flow rate may be used, dependent of the internal diameter of the columns used or the instructions of the manufacturer of the column.
Izbira kolon(-e) lahko bistveno vpliva na retenzijski čas posameznih vrhov.
Note 2.
Integrator (6.11.6) samodejno izračuna površino A vsakega vrha:
Rinse the columns with water during each interruption.
površina vrha II
Never leave the eluent solution in them (5.2).
površina vrha III
Prior to any interruption of more than 24 hours, rinse the columns with water then wash them with solution (5.3) for at least three hours at a flow rate of 0,2 ml per minute.
Pred kvantitativno interpretacijo je treba pregledati videz vsakega kromatograma, da bi tako lahko izsledili kakršne koli nepravilnosti kot posledice slabega delovanja bodisi kromatografa ali kolon ali pa kot posledico izvora in narave analiziranega vzorca.
The results of chromatographic analysis of the test sample [E] are obtained in the form of chromatogram in which each peak is identified by its retention time RT as follows:
V primeru dvoma analizo ponovite.
Peak II:
Kalibracija
The second peak of the chromatogram having an RT of about 12,5 minutes.
Za standardne vzorce (5.4) uporabite postopek, opisan od točke 8.2 do točke 8.4.2.
Peak III:
Uporabite sveže pripravljene raztopine, ker se CMP razgradi v 8-odstotnem trikloroocetnem okolju.
The third peak of the chromatogram, corresponding to the CMP, having an RT of 15,5 minutes.
Izguba je ocenjena na 0,2 % na uro pri 30 °C.
The choice of the column(s) can affect the retention times of the individual peaks considerably.
Pred kromatografskim določanjem vzorcev pripravite kolone s ponavljajočim vbrizgavanjem standardnega vzorca (5.4.2) v raztopino (8.5.1), dokler nista površina in retenzijski čas vrha, ki ustreza CMP, stalna.
The integrator (6.11.6) automatically calculates the area A of each peak:
Z vbrizganjem enake prostornine filtrata (8.5.1), kakor je uporabljen pri vzorcih, določite odzivne faktorje R.
area of peak II,
IZRAŽANJE REZULTATOV
area of peak III,
Metoda za izračun in enačbe