natgillin/translations-raw huggingface dataset:
Negative, i.e., not more than 350 mU of phosphatasic activity per litre of reconstituted milk
Najmanje 34,0 % u pogledu nemasne suhe tvari
Insolubility index
Najviše 1,00 %
Maximum 0,5 ml (24 °C)
Najviše 3,5 %
Scorched particles
Titracijska kiselost u ml decinormalne otopine natrijeva hidroksida
Maximum 15,0 mg, i.e. disc B minimum
Najviše 19,5 ml
Micro-organisms
Laktati
Maximum 40000 CFU per gram
Najviše 150 mg/100 g
Buttermilk
Pokus na fosfatazu
None
Negativan, tj. nema više od 350 mU fosfatazne aktivnosti po litri rekonstituiranog mlijeka
Rennet whey
Indeks netopivosti
Acid whey
Najviše 0,5 ml (24 °C)
None or maximum 150 mg/100 g
Sagorjele čestice
Taste and smell
Najviše 15,0 mg, tj. minimalno disk B
Clean
Mikroorganizmi
Appearance
Najviše 40000 CFU/g
White or slightly yellowish colour, free from impurities and coloured particles
Mlaćenica
“Buttermilk” means the by-product of butter production obtained after churning of the cream and separation of the solid fat.
Nema je
The absence of buttermilk can be established either by an on-the-spot inspection of the production plant carried out without prior notice at least once a week, or by a laboratory analysis of the end product indicating a maximum of 69,31 mg of PEDP phosphatidylethanolamine dipalmitoyl per 100 g.
Sirutka dobivena sirištem
“Whey” means the by-product of cheese or casein production obtained by the action of acids, rennet and/or chemico-physical processes.
Sirutka dobivena kiselinom
Having regard to Regulation (EU) No 1306/2013 of the European Parliament and of the Council of 17 December 2013 on the financing, management and monitoring of the common agricultural policy and repealing Council Regulations (EEC) No 352/78, (EC) No 165/94, (EC) No 2799/98, (EC) No 814/2000, (EC) No 1290/2005 and (EC) No 485/2008, and in particular Article 62(2)(i) thereof,
uzimajući u obzir Uredbu (EU) br.
Whereas:
1306/2013 Europskog parlamenta i Vijeća od 17. prosinca 2013. o financiranju, upravljanju i nadzoru zajedničke poljoprivredne politike i o stavljanju izvan snage uredaba Vijeća (EEZ) br.
Commission Delegated Regulation (EU) 2016/1238 and Commission Implementing Regulation (EU) 2016/1240 lay down the rules on public intervention and aid for private storage.
352/78, (EZ) br.
Commission Regulation (EC) No 273/2008 sets out the methods to be applied in assessing whether milk and milk products comply with the eligibility requirements laid down in those Regulations for public intervention and aid for private storage.
165/94, (EZ) br.
In the light of technical developments in the methodology used in the analysis and quality evaluation of milk and milk products, substantial changes should be made in order to simplify and to provide for updated references to ISO standards.
2799/98, (EZ) br.
In the interests of clarity and efficiency, and having regard to the extent and technical nature of the amendments to the provisions of Regulation (EC) No 273/2008, the relevant provisions of that Regulation should be incorporated into Implementing Regulation (EU) 2016/1240.
814/2000, (EZ) br.
In order to ensure uniform compliance with the new standards and methods across Member States, laboratories should be allowed a sufficient period of time to review procedures and apply the updated methods.
1290/2005 i (EZ) br.
Implementing Regulation (EU) 2016/1240 should therefore be amended accordingly.
485/2008, a posebno njezin članak 62. stavak 2. točku (i),
In the interests of legal certainty Regulation (EC) No 273/2008 should be repealed.
budući da:
The measures provided for in this Regulation are in accordance with the opinion of the Committee for the Common Organisation of the Agricultural Markets,
Delegiranom uredbom Komisije (EU) 2016/1238 i Provedbenom uredbom Komisije (EU) 2016/1240 utvrđuju se pravila o javnoj intervenciji i potporama za privatno skladištenje.
HAS ADOPTED THIS REGULATION:
Uredbom Komisije (EZ) br.
Article 1
273/2008 utvrđuju se metode koje se primjenjuju kako bi se ocijenilo ispunjavaju li mlijeko i mliječni proizvodi uvjete prihvatljivosti utvrđene u navedenim uredbama u vezi s javnom intervencijom i privatnim skladištenjem.
Implementing Regulation (EU) 2016/1240 is amended as follows:
S obzirom na tehnički napredak u području metodologije koja se upotrebljava u analizi i ocjenjivanju kvalitete mlijeka i mliječnih proizvoda trebalo bi uvesti znatne promjene radi pojednostavnjenja i ažuriranja upućivanja na norme ISO.
Article 4 is amended as follows:
Radi osiguranja jasnoće i učinkovitosti te uzimajući u obzir opseg i tehničku prirodu izmjena odredbi Uredbe (EZ) br.
paragraph 1 is amended as follows:
273/2008 relevantne odredbe te uredbe trebalo bi uključiti u Provedbenu uredbu (EU) 2016/1240.
point (d) is replaced by the following:
Kako bi se osiguralo jedinstveno poštovanje novih normi i metoda u svim državama članicama, laboratorijima bi trebalo osigurati dovoljno vremena za preispitivanje postupaka i primjenu ažuriranih metoda.
for butter: in Parts I and Ia of Annex IV to this Regulation’;
Provedbenu uredbu (EU) 2016/1240 trebalo bi stoga na odgovarajući način izmijeniti.
point (e) is replaced by the following:
U interesu pravne sigurnosti Uredbu (EZ) br.
for skimmed milk powder: in Parts I and Ia of Annex V to this Regulation’;
273/2008 trebalo bi staviti izvan snage.
paragraph 2 is replaced by the following:
Mjere predviđene ovom Uredbom u skladu su s mišljenjem Odbora za zajedničku organizaciju poljoprivrednih tržišta,
The methods to be used to determine the quality of cereals, butter and skimmed milk powder eligible for public intervention referred to in Annexes I, IV and V respectively, shall be those established by the latest versions of the relevant European or international standards, as the case may be, in force at least 6 months before the first day of the public intervention period as defined in Article 12 of Regulation (EU) No 1308/2013.’;
DONIJELA JE OVU UREDBU:
the following Article 60a is inserted:
Članak 1.
‘Article 60a
Provedbena uredba (EU) 2016/1240 mijenja se kako slijedi:
Specific provision on checks relating to public intervention and aid for private storage for milk and milk products
1.
The eligibility of butter, skimmed milk powder and cheese to receive aid for private storage shall be established in accordance with the methods laid down in Annexes VI, VII and VIII respectively.
Članak 4. mijenja se kako slijedi:
Those methods shall be established by reference to the latest versions of the relevant European or international standards, as the case may be, in force at least 6 months before the first day of the public intervention period as defined in Article 12 of Regulation (EU) No 1308/2013.
stavak 1. mijenja se kako slijedi:
The results of the checks conducted by applying the methods set out in this Regulation shall be evaluated in accordance with Annex IX.’;
i. točka (d) zamjenjuje se sljedećim:
the Annexes are amended in accordance with the Annex to this Regulation.
za maslac: dijelovima I. i I.a Priloga IV. ovoj Uredbi”;
Article 2
ii. točka (e) zamjenjuje se sljedećim:
Regulation (EC) No 273/2008 is repealed.
za obrano mlijeko u prahu: dijelovima I. i I.a Priloga V. ovoj Uredbi”;
Article 3
stavak 2. zamjenjuje se sljedećim:
This Regulation shall enter into force on the seventh day following that of its publication in the Official Journal of the European Union.
Metode za određivanje kvalitete žitarica, maslaca i obranog mlijeka u prahu prihvatljivih za javnu intervenciju koje su navedene u prilozima I., IV. odnosno V. jesu metode utvrđene najnovijim verzijama relevantnih europskih ili međunarodnih normi, ovisno o slučaju, koje su na snazi najmanje 6 mjeseci prije prvog dana razdoblja javne intervencije kako je utvrđeno u članku 12.
Done at Brussels, 30 January 2018.
1308/2013.”.
For the Commission
Umeće se sljedeći članak 60.a:
The President
„Članak 60.a
OJ L 347, 20.12.2013, p.
Posebne odredbe o kontrolama koje se odnose na javnu intervenciju i potpore za privatno skladištenje za mlijeko i mliječne proizvode
549.
Prihvatljivost maslaca, obranog mlijeka u prahu i sira za primanje potpore za privatno skladištenje utvrđuje se u skladu s metodama iz priloga VI., VII. odnosno VIII.
Commission Delegated Regulation (EU) 2016/1238 of 18 May 2016 supplementing Regulation (EU) No 1308/2013 of the European Parliament and of the Council with regard to public intervention and aid for private storage (OJ L 206, 30.7.2016, p.
Te se metode utvrđuju uzimajući u obzir najnovije verzije relevantnih europskih ili međunarodnih normi, ovisno o slučaju, koje su na snazi najmanje 6 mjeseci prije prvog dana razdoblja javne intervencije kako je utvrđeno u članku 12.
15).
Uredbe (EU) br.
Commission Implementing Regulation (EU) 2016/1240 of 18 May 2016 laying down rules for the application of Regulation (EU) No 1308/2013 of the European Parliament and of the Council with regard to public intervention and aid for private storage (OJ L 206, 30.7.2016, p.
1308/2013.
71).
Rezultati kontrola provedenih primjenom metoda utvrđenih u ovoj Uredbi ocjenjuju se u skladu s Prilogom IX.”.
Commission Regulation (EC) No 273/2008 of 5 March 2008 laying down detailed rules for the application of Council Regulation (EC) No 1255/1999 as regards methods for the analysis and quality evaluation of milk and milk products (OJ L 88, 29.3.2008, p.
Prilozi se mijenjaju u skladu s Prilogom ovoj Uredbi.
The Annexes to Implementing Regulation (EU) 2016/1240 are amended as follows:
273/2008 stavlja se izvan snage.
Annex IV is amended as follows:
Članak 3.
in Part I, point 2, the second sub-paragraph is replaced by the following:
Ova Uredba stupa na snagu sedmog dana od dana objave u Službenom listu Europske unije.
‘Each sample shall be assessed individually.
Ova je Uredba u cijelosti obvezujuća i izravno se primjenjuje u svim državama članicama.
No resampling or re-evaluation is allowed.’;
Sastavljeno u Bruxellesu 30. siječnja 2018.
the following Part Ia is inserted:
Za Komisiju
‘PART IA
Predsjednik
Methods of analysis of unsalted butter for public intervention
SL L 347, 20.12.2013., str.
Parameter
549.
Method
Delegirana uredba Komisije (EU) 2016/1238 od 18. svibnja 2016. o dopuni Uredbe (EU) br.
ISO 17189 or ISO 3727 part 3
15.)
Water
Provedbena uredba Komisije (EU) 2016/1240 od 18. svibnja 2016. o utvrđivanju pravila za primjenu Uredbe (EU) br.
ISO 3727 part 1
1308/2013 Europskog parlamenta i Vijeća s obzirom na javnu intervenciju i potpore za privatno skladištenje (SL L 206, 30.7.2016., str.
Non-fat solids
71.)
ISO 3727 part 2
Uredba Komisije (EZ) br.
Fat acidity
273/2008 od 5. ožujka 2008. o utvrđivanju detaljnih pravila za primjenu Uredbe Vijeća (EZ) br.
Peroxide value
1255/1999 o metodama analize i ocjenjivanja kvalitete mlijeka i mliječnih proizvoda (SL L 88, 29.3.2008., str.
Non-milk fat
1.).
Sensory characteristics
PRILOG
ISO 22935 parts 2 and 3 and scoring table hereafter.
Prilozi Provedbenoj uredbi (EU) 2016/1240 mijenjaju se se kako slijedi:
Scoring table
1.
Appearance
Prilog IV. mijenja se kako slijedi:
Consistency
u dijelu I. točki 2. drugi podstavak zamjenjuje se sljedećim:
Odour and Flavour
„Svaki se uzorak ocjenjuje pojedinačno.
Points
Nije dopušteno ponovno uzimanje uzoraka ili ponovno ocjenjivanje.”
Remarks
umeće se sljedeći dio I.a:
Very good
„DIO I.A
Ideal type
Metode analize nesoljenog maslaca za javnu intervenciju
Highest quality
Parametar
(equal dry)
Metoda
(equal spreadable)
Masti
(absolutely pure finest odour)
ISO 17189 ili ISO 3727 dio 3
Good
Voda
(no evident defects)
ISO 3727 dio 1
1, 2 or 3
Bezmasna suha tvar
Any defect
ISO 3727 dio 2
Any defect’
Kiselost masti
in Annex V the following Part Ia is inserted:
Peroksidni broj
Methods of analysis of skimmed milk powder for public intervention
Nemliječna mast
Protein
Senzorna svojstva
ISO 8968 part 1
ISO 22935 dijelovi 2 i 3 i tablica bodovanja u nastavku.
Phosphatase test
Konzistencija
ISO 11816 part 1
Miris i okus
Insolubility index
Bodovi
ISO 8156
Napomene
Scorched particles ADPI
Vrlo dobri
Micro-organisms
idealni tip
ISO 4833-part 1
najviša kvaliteta
Buttermilk
(ravnomjerna suhoća)
Appendix I
(ravnomjerna mazivost)
Rennet whey
(potpuno čist, blagi miris)
Appendix II and III
Dobri
Acid whey
(bez očitih nedostataka)
ISO 8069 or On-the-spot inspections
1, 2 ili 3
Sensory checks
Nedostatci
ISO 22935 part 2 and 3
Nedostatci”
SKIMMED MILK POWDER: QUANTITATIVE DETERMINATION OF PHOSPHATIDYLSERINE AND PHOSPHATIDYLETHANOLAMINE
U Prilogu V. umeće se sljedeći dio I.a:
Method: reversed-phase HPLC
Metode analize obranog mlijeka u prahu za javnu intervenciju
PURPOSE AND FIELD OF APPLICATION
Bjelančevine
The method describes a procedure for the quantitative determination of phosphatidylserine (PS) and phosphatidylethanolamine (PE) in skimmed milk powder (SMP) and is suitable for detecting buttermilk solids in SMP.
ISO 8968 dio 1
DEFINITION
Kiselost
PS + PE content : the mass fraction of substance determined using the procedure here specified.
Laktati
The result is expressed as milligrams of phosphatidylethanolamine dipalmitoyl (PEDP) per 100 g powder.
Pokus na fosfatazu
PRINCIPLE OF THE METHOD
ISO 11816 dio 1
Extraction of aminophospholipids by methanol from reconstituted milk powder.
Indeks netopivosti
Determination of PS and PE as o-phthaldialdehyde (OPA) derivatives by reversed-phase (RP) HPLC and fluorescence detection.
Mikroorganizmi
Quantification of PS and PE content in the test sample by reference to a standard sample containing a known amount of PEDP.
ISO 4833-dio 1
REAGENTS
Mlaćenica
All reagents shall be of recognised analytical grade.
Dodatak I.
Water shall be distilled or water of at least equivalent purity, unless otherwise specified.
Slatka sirutka
Standard material: PEDP, at least 99 % pure
Dodatci II. i III.
Note: Standard material shall be stored at – 18 °C.
Kisela sirutka
Reagents for standard sample and test sample preparation
ISO 8069 ili kontrole na terenu
HPLC-grade methanol
Senzorne kontrole
HPLC-grade chloroform
ISO 22935 dijelovi 2 i 3
Tryptamine-monohydrochloride
OBRANO MLIJEKO U PRAHU: KVANTITATIVNO ODREĐIVANJE FOSFATIDILSERINA I FOSFATIDILETANOLAMINA
Reagents for o-phthaldialdehyde derivatisation
Metoda: reverzno-fazna HPLC metoda
Sodium hydroxide, 12 M water solution
SVRHA I PODRUČJE PRIMJENE
Boric acid, 0,4 M water solution adjusted to pH 10,0 with sodium hydroxide (4.3.1)
Ova metoda opisuje postupak količinskog određivanja fosfatidilserina (PS) i fosfatidiletanolamina (PE) u obranom mlijeku u prahu (OMP) te je prikladna za detekciju suhe tvari mlaćenice u OMP-u.
2-mercaptoethanol
DEFINICIJA
o-phthaldialdehyde (OPA)
Sadržaj fosfatidilserina (PS) i fosfatidiletanolamina (PE) : masena frakcija određene tvari koja se utvrđuje ovdje opisanim postupkom.
HPLC elution solvents
Rezultat se izražava u fosfatidiletanolamin dipalmitoilu (PEDP) na 100 g praha.
Elution solvents shall be prepared using HPLC-grade reagents.
NAČELO METODE
HPLC-grade water
Ekstrakcija aminofosfolipida iz rekonstituiranog mlijeka u prahu pomoću metanola.
Methanol of tested fluorimetric purity
Određivanje fosfatidilserina (PS) i fosfatidiletanolamina (PE) kao derivata o-ftaldialdehida (OPA) reverzno-faznom (RP) HPLC analizom i detekcijom fluorescencije.
Tetrahydrofuran
Kvantifikacija sadržaja fosfatidilserina (PS) i fosfatidiletanolamina (PE) u ispitnom uzorku na temelju standardnog uzorka koji sadržava poznatu količinu fosfatidiletanolamin dipalmitoila (PEDP).
Sodium dihydrogen phosphate
REAGENSI
Sodium acetate
Svi reagensi moraju biti priznate analitičke čistoće.
Acetic acid.
Mora se koristiti destilirana voda ili voda barem jednake čistoće, osim ako je navedeno drukčije.
APPARATUS
Standardni materijal: fosfatidiletanolamin dipalmitoil (PEDP), čistoće najmanje 99 %
Analytical balance, capable of weighing to the nearest 1 mg, with a readability of 0,1 mg
Napomena: Standardni materijal čuva se na temperaturi od – 18 °C.
Beakers, 25 and 100 ml capacity
Reagensi za pripremu standardnog i ispitnog uzorka
Pipettes, capable of delivering 1 and 10 ml
Metanol, HPLC čistoće
Magnetic stirrer
Kloroform, HPLC čistoće
Graduated pipettes, capable of delivering 0,2, 0,5 and 5 ml
Triptamin-monohidroklorid
Volumetric flasks, 10, 50 and 100 ml capacity
Reagensi za derivatizaciju o-ftaldialdehida
Syringes, 20 and 100 μl capacity
Natrijev hidroksid, vodena otopina 12 M
Ultrasonic bath
Borna kiselina, vodena otopina 0,4 M, pH vrijednost podešena na 10,0 pomoću natrijeva hidroksida (4.3.1.)
Centrifuge, capable of operating at 27000 × g
2-merkaptoetanol
Glass vials, about 5 ml capacity
O-ftaldialdehid (OPA)
Graduated cylinder, 25 ml capacity
Otapala za HPLC eluaciju
pH-meter, accurate to 0,1 pH units
Za pripremu otapala za eluaciju koriste se reagensi HPLC čistoće
HPLC equipment
Voda HPLC čistoće
Gradient pumping system, capable of operating at 1,0 ml/min at 200 bar
Metanol provjerene fluorimetrijske čistoće
Autosampler with derivatisation capability
Tetrahidrofuran
Column heater, capable of maintaining the column at 30 °C ± 1 °C
Natrijev dihidrogen fosfat
Fluorescence detector, capable of operating at 330 nm excitation wavelength and 440 nm emission wavelength
Natrijev acetat
Integrator or data processing software capable of peak area measurement
Octena kiselina.
A LiChrospher® — 100 column (250 × 4,6 mm) or an equivalent column packed with octadecylsilane (C 18), 5 μm particle size.
OPREMA
SAMPLING
Analitička vaga koja može vagati s točnošću od 1 mg, uz mogućnost očitanja 0,1 mg.
Sampling shall be carried out in accordance with ISO Standard 707.
Laboratorijske čaše zapremine 25 i 100 ml
PROCEDURE
Pipete za prijenos količina od 1 i 10 ml
Preparation of the internal standard solution
Magnetska miješalica
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)
Graduirane pipete za prijenos količina od 0,2, 0,5 i 5 ml
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
Odmjerne tikvice zapremine 10, 50 i 100 ml
Preparation of the test sample solution
Šprice zapremine 20 i 100 μl
Weigh 1,000 ± 0,001 g of the SMP sample into a 25 ml beaker (5.2).
Ultrazvučna kupelj
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
Centrifuga koja može raditi na 27000 × g
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).
Staklene bočice zapremine oko 5 ml
Mix carefully by inversion and sonicate (5.8) for 15 min
Menzura zapremine 25 ml
Centrifuge (5.9) at 27000 g × g for 10 minutes and collect the supernatant in a glass vial (5.10)
pH-metar s točnošću od 0,1 pH jedinica
Note: Test sample solution should be stored at 4 °C until the HPLC analysis is performed.
Oprema za HPLC
Preparation of the external standard solution
Gradijentni pumpni sustav s mogućnošću rada pri brzini protoka od 1,0 ml/min i tlaku od 200 bara
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).
Automatski uređaj za uzorkovanje s mogućnošću derivatizacije
Heat the stoppered flask to 50 °C ± 1 °C and mix carefully till the PEDP dissolves.
Grijač kolona koji kolonu može održavati na temperaturi 30 °C ± 1 °C
Cool the flask to 20 °C, make up to the volume with methanol (4.2.1) and mix by inversion
Detektor fluorescencije s mogućnošću rada na 330 nm valne duljine ekscitacije i 440 nm valne duljine emisije
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).
Integrator ili softver za obradu podataka koji može izmjeriti površinu pika
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).
Kolona LiChrospher® – 100 (250 × 4,6 mm) ili ekvivalentna kolona ispunjena oktadecilsilanom (C 18), veličine čestica 5 μm
Mix by inversion
UZORKOVANJE
Note: Reference sample solution should be stored at 4 °C until the HPLC analysis is performed.
Uzorkovanje se provodi u skladu s normom ISO 707.
Preparation of the derivatising reagent
POSTUPAK
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.
Priprema interne standardne otopine
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).
Odvagnite 30,0 ± 0,1 mg triptamin-monohidroklorida (4.2.3.) u odmjernu tikvicu od 100 ml (5.6.) i dopunite do oznake metanolom (4.2.1.).
Note: The derivatising reagent should be stored at 4 °C in a brown glass vial and is stable for one week.
Otpipetirajte 1 ml (5.3.) ove otopine u odmjernu tikvicu od 10 ml (5.6.) i dopunite do oznake metanolom (4.2.1.) da biste dobili koncentraciju triptamina od 0,15 mM.
Determination by HPLC
Priprema otopine ispitnog uzorka
Elution solvents (4.4)
Odvagnite 1,000 ± 0,001 g uzorka OMP-a u laboratorijsku čašu od 25 ml (5.2.).
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)
Pipetom (5.3.) dodajte 10 ml destilirane vode temperature 40 ± 1 °C i miješajte 30 minuta magnetskom miješalicom (5.4.) da bi se rastopile eventualne grudice.
Solvent B: methanol
Otpipetirajte 0,2 ml (5.5.) rekonstituiranog mlijeka u odmjernu tikvicu od 10 ml (5.6.), špricom (5.7.) dodajte 100 μl otopine triptamina koncentracije 0,15 mM (7.1.) i dopunite do oznake metanolom (4.2.1.).
Suggested eluting gradient:
Pažljivo izmiješajte preokretanjem bočice i stavite na 15 minuta u ultrazvučnu kupelj (5.8.).
Time (min)
Centrifugirajte (5.9.) na 27000 × g 10 minuta i skupite supernatant u staklenu bočicu (5.10.).
Solvent A (%)
Napomena: Otopinu ispitnog uzorka čuvajte na temperaturi od 4 °C do izvođenja HPLC analize.
Solvent B (%)
Priprema vanjske standardne otopine
Flow rate (ml/min)
Odvagnite 55,4 mg fosfatidiletanolamin dipalmitoila (PEDP) (4.1.) u odmjernu tikvicu od 50 ml (5.6.) i dodajte oko 25 ml kloroforma (4.2.2.) pomoću menzure (5.11.).
Initial
Zagrijte začepljenu tikvicu na 50 °C ± 1 °C i pažljivo miješajte da se PEDP rastopi.
0Note: The eluting gradient may require slight modification in order to achieve the resolution shown in figure 1.
Ohladite tikvicu na 20 °C, dopunite do oznake metanolom (4.2.1.) i izmiješajte preokretanjem tikvice.
Column temperature: 30 °C.
Otpipetirajte 1 ml (5.3.) ove otopine u odmjernu tikvicu od 100 ml (5.6.) i dopunite do oznake metanolom (4.2.1.).
Injection volume: 50 μl derivatising reagent and 50 μl sample solution
Otpipetirajte 1 ml (5.3.) tako dobivene otopine u odmjernu tikvicu od 10 ml (5.6.), dodajte 100 μl (5.7.) otopine triptamina koncentracije 0,15 mM (7.1.) i dopunite do oznake metanolom (4.2.1.).
Column equilibration
Promiješajte preokretanjem tikvice.
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.
Napomena: Otopinu referentnog uzorka do HPLC analize treba čuvati na temperaturi od 4 °C.
Perform a blank run by injecting methanol (4.2.1).
Priprema reagensa za derivatizaciju
Note: Before long-term storage flush the column with methanol: chloroform = 80:20 (v/v) for 30 minutes.
Odvagnite 25,0 ± 0,1 mg o-ftaldialdehida (OPA) (4.3.4.) u odmjernu tikvicu zapremine 10 ml (5.6.), dodajte 0,5 ml (5.5.) metanola (4.2.1.) i pažljivo promiješajte da se OPA rastopi.
Determine the PS + PE content in the test sample
Dopunite do oznake otopinom borne kiseline (4.3.2.) i špricom (5.7.) dodajte 20 μl 2-merkaptoetanola (4.3.3.).
Perform the sequence of the chromatographic analyses keeping constant the run-to-run time in order to obtain constant retention times.
Napomena: Reagens za derivatizaciju treba čuvati na 4 °C u tamnoj staklenoj bočici da bi ostao stabilan tjedan dana.
Inject the external standard solution (7.3) every 5-10 test sample solutions in order to calculate the response factor
Određivanje putem HPLC-a