SIST EN 16167:2018
(Main)Soil, treated biowaste and sludge - Determination of polychlorinated biphenyls (PCB) by gas chromatography with mass selective detection (GC-MS) and gas chromatography with electron-capture detection (GC-ECD)
Soil, treated biowaste and sludge - Determination of polychlorinated biphenyls (PCB) by gas chromatography with mass selective detection (GC-MS) and gas chromatography with electron-capture detection (GC-ECD)
This draft European Standard specifies a method for quantitative determination of seven selected polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in sludge, treated biowaste and soil using GC-MS and GC-ECD (see Table 2).
(...)
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this European Standard, limit of application of 1 µg/kg (expressed as dry matter) can be achieved.
Sludge and treated biowaste may differ in properties and also in the expected contamination levels of PCBs and presence of interfering substances. These differences make it impossible to describe one general procedure. This European Standard contains decision tables based on the properties of the sample and the extraction and clean-up procedure to be used.
Boden, behandelter Bioabfall und Schlamm - Bestimmung von polychlorierten Biphenylen (PCB) mittels Gaschromatographie mit Massenspektrometrie-Kopplung (GC-MS) und Gaschromatographie mit Elektroneneinfangdetektion (GC-ECD)
Der vorliegende Europäische Norm Entwurf legt ein Verfahren für die quantitative Bestimmung von sieben ausgewählten polychlorierten Biphenylen (PCB 28, PCB 52, PCB 101, PCB 118, PCB 138, PCB 153 und PCB 180) in Schlamm, behandeltem Bioabfall und Boden unter Anwendung der GC MS und GC ECD fest (siehe Tabelle 2).
(...)
Die Nachweisgrenze hängt von den zu bestimmenden Substanzen, den verwendeten Geräten, der Qualität der für die Extraktion der Probe verwendeten Chemikalien und der Reinigung des Extrakts ab.
Unter den in dieser Europäischen Norm festgelegten Bedingungen kann eine Anwendungsgrenze von 1 µg/kg (angegeben als Trockenmasse) erreicht werden.
Schlamm und behandelter Bioabfall können sich in ihren Eigenschaften sowie in den erwarteten Kontaminationsniveaus von PCB und der Anwesenheit von störenden Substanzen unterscheiden. Aufgrund dieser Unterschiede ist es unmöglich, ein allgemeines Verfahren darzulegen. Die vorliegende Europäische Norm enthält Entscheidungstabellen, die auf den Probeneigenschaften und den anzuwendenden Extraktions und Reinigungsverfahren basieren.
Sols, biodéchets traités et boues - Dosage des polychlorobiphényles (PCBs) par chromatographie en phase gazeuse-spectrométrie gazeuse couplée avec un détecteur de masse (CG-SM) ou un détecteur par capture d'électrons (CG-ECD)
La présente Norme européenne spécifie une méthode de dosage, par CG-SM et CG-ECD, de sept polychlorobiphényles choisis (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 et PCB180) dans les boues, les biodéchets traités et les sols (voir Tableau 2).
(...)
La limite de détection dépend des composés à analyser, de l’équipement utilisé, de la qualité des réactifs chimiques utilisés pour l’extraction de l’échantillon et la purification de l’extrait.
Dans les conditions spécifiées dans la présente Norme européenne, une limite d'application de 1 µg/kg (exprimée sur matière sèche) peut être atteinte.
Les boues et les biodéchets traités peuvent présenter des différences en termes de propriétés, de niveaux prévus de contamination par les PCBs et de présence de substances interférentes. En raison de ces différences, il est impossible de décrire un mode opératoire général. La présente Norme européenne contient des tables de décision basées sur les propriétés de l'échantillon ainsi que sur le mode opératoire d'extraction et de purification à utiliser.
Tla, obdelani biološki odpadki in blato - Določevanje polikloriranih bifenilov (PCB) s plinsko kromatografijo z masno selektivnim detektorjem (GC/MS) in s plinsko kromatografijo z detektorjem z zajetjem elektronov (GC/ECD)
Ta osnutek evropskega standarda določa metodo za kvantitativno določevanje sedmih izbranih polikloriranih bifenilov (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 in PCB180) v blatu, obdelanih bioloških odpadkih ter tleh z metodama GC-MS in GC-ECD (glej preglednico 2).
(...)
Meja detekcije je odvisna od determinant, uporabljene opreme, kakovosti kemikalij, uporabljenih za ekstrakcijo vzorca in očiščenje izvlečka.
Pod pogoji, določenimi v tem evropskem standardu, se lahko doseže meja uporabe 1 μg/kg (izraženo kot suha snov).
Blato in obdelani biološki odpadki se lahko razlikujejo glede lastnosti, predvidenih stopenj onesnaženja s polikloriranimi bifenili ter prisotnosti motečih snovi. Zaradi teh razlik ni možen opis enotnega splošnega postopka. Ta evropski standard vključuje tabele odločanja, ki temeljijo na lastnostih vzorca, ter postopek za ekstrakcijo in očiščenje, ki ga je treba uporabiti.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN 16167:2018
01-december-2018
1DGRPHãþD
SIST EN 16167:2013
7ODREGHODQLELRORãNLRGSDGNLLQEODWR'RORþHYDQMHSROLNORULUDQLKELIHQLORY 3&%
VSOLQVNRNURPDWRJUDILMR]PDVQRVHOHNWLYQLPGHWHNWRUMHP *&06 LQVSOLQVNR
NURPDWRJUDILMR]GHWHNWRUMHP]]DMHWMHPHOHNWURQRY *&(&'
Soil, treated biowaste and sludge - Determination of polychlorinated biphenyls (PCB) by
gas chromatography with mass selective detection (GC-MS) and gas chromatography
with electron-capture detection (GC-ECD)
Boden, behandelter Bioabfall und Schlamm - Bestimmung von polychlorierten
Biphenylen (PCB) mittels Gaschromatographie mit Massenspektrometrie-Kopplung (GC-
MS) und Gaschromatographie mit Elektroneneinfangdetektion (GC-ECD)
Sols, biodéchets traités et boues - Dosage des polychlorobiphényles (PCBs) par
chromatographie en phase gazeuse-spectrométrie gazeuse couplée avec un détecteur
de masse (CG-SM) ou un détecteur par capture d'électrons (CG-ECD)
Ta slovenski standard je istoveten z: EN 16167:2018
ICS:
13.030.20 7HNRþLRGSDGNL%ODWR Liquid wastes. Sludge
13.080.10 .HPLMVNH]QDþLOQRVWLWDO Chemical characteristics of
soils
71.040.50 Fizikalnokemijske analitske Physicochemical methods of
metode analysis
SIST EN 16167:2018 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN 16167:2018
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SIST EN 16167:2018
EN 16167
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2018
EUROPÄISCHE NORM
ICS 13.030.01; 13.080.10 Supersedes EN 16167:2012
English Version
Soil, treated biowaste and sludge - Determination of
polychlorinated biphenyls (PCB) by gas chromatography
with mass selective detection (GC-MS) and gas
chromatography with electron-capture detection (GC-
ECD)
Sols, biodéchets traités et boues - Dosage des Boden, behandelter Bioabfall und Schlamm -
polychlorobiphényles (PCBs) par chromatographie en Bestimmung von polychlorierten Biphenylen (PCB)
phase gazeuse-spectrométrie gazeuse couplée avec un mittels Gaschromatographie mit
détecteur de masse (CG-SM) ou un détecteur par Massenspektrometrie-Kopplung (GC-MS) und
capture d'électrons (CG-ECD) Gaschromatographie mit Elektroneneinfangdetektion
(GC-ECD)
This European Standard was approved by CEN on 20 March 2018.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2018 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 16167:2018 E
worldwide for CEN national Members.
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SIST EN 16167:2018
EN 16167:2018 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 5
3 Terms and definitions . 6
4 Principle . 7
5 Interferences . 7
5.1 Interference with sampling and extraction . 7
5.2 Interference with GC . 7
6 Safety remarks . 8
7 Reagents . 8
8 Apparatus . 14
9 Sample storage and preservation . 15
9.1 Sample storage . 15
9.2 Sample pretreatment . 15
10 Procedure. 16
10.1 Blank test . 16
10.2 Extraction . 16
10.3 Concentration . 18
10.4 Clean-up of the extract . 19
10.5 Addition of the injection standard . 22
10.6 Gas chromatographic analysis (GC) . 22
10.7 Mass spectrometry (MS) . 23
10.8 Electron capture detection (ECD) . 28
11 Performance characteristics . 30
12 Precision . 30
13 Test report . 30
Annex A (informative) Repeatability and reproducibility data . 31
Annex B (informative) Examples for retention times of PCBs . 33
Annex C (informative) Calculation method for the estimation of total PCB content . 34
Bibliography . 39
2
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SIST EN 16167:2018
EN 16167:2018 (E)
European foreword
This document (EN 16167:2018) has been prepared by Technical Committee CEN/TC 444 “Test
methods for environmental characterization of solid matrices”, the secretariat of which is held by NEN.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by February 2019, and conflicting national standards
shall be withdrawn at the latest by February 2019.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document will supersede EN 16167:2012.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
The preparation of this document by CEN is based on a mandate by the European Commission
(Mandate M/330), which assigned the development of standards on sampling and analytical methods
for hygienic and biological parameters as well as inorganic and organic determinants, aiming to make
these standards applicable to sludge, treated biowaste and soil as far as this is technically feasible.
According to the CEN-CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria,
Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta,
Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,
Turkey and the United Kingdom.
3
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SIST EN 16167:2018
EN 16167:2018 (E)
Introduction
Polychlorinated biphenyls (PCB) have been widely used as additives in industrial applications where
chemical stability has been required. This stability on the other hand creates environmental problems
when PCBs are eventually released into the environment. Since some of these PCB compounds are
highly toxic, their presence in the environment (air, water, soil, sediment and waste) is regularly
monitored and controlled. At present determination of PCB is carried out in these matrices in most of
the routine laboratories following the preceding steps for sampling, pretreatment, extraction, clean-up
by measurement of specific PCB by means of gas chromatography in combination with mass
spectrometric detection (GC-MS) or gas chromatography with electron capture detector (GC-ECD).
This European Standard was developed in the European project 'HORIZONTAL'. It is the result of a desk
study “3-12 PCB” and aims at evaluation of the latest developments in assessing PCBs in sludge, soil,
treated biowaste and neighbouring fields. Taken into account the different matrices and possible
interfering compounds, this European Standard does not contain one single possible way of working.
Several choices are possible, in particular relating to clean-up. Detection with both MS-detection and
ECD-detection is possible. Three different extraction procedures are described and 11 clean-up
procedures. The use of internal and injection standards is described in order to have an internal check
on choice of the extraction and clean-up procedure. The method is as far as possible in agreement with
the method described for PAHs (see EN 16181). It has been tested for ruggedness.
This European Standard is applicable and validated for several types of matrices as indicated in Table 1
(see also Annex A for the results of the validation).
Table 1 — Matrices for which this European Standard is applicable and validated
Matrix Materials used for validation
Sludge Municipal sewage sludge
Biowaste Compost
Soil Sandy soil
WARNING — Persons using this European Standard should be familiar with usual laboratory practice.
This European Standard does not purport to address all of the safety problems, if any, associated with
its use. It is the responsibility of the user to establish appropriate safety and health practices and to
ensure compliance with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted according to this European
Standard be carried out by suitably trained staff.
4
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SIST EN 16167:2018
EN 16167:2018 (E)
1 Scope
This draft European Standard specifies a method for quantitative determination of seven selected
polychlorinated biphenyls (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153 and PCB180) in sludge,
treated biowaste and soil using GC-MS and GC-ECD (see Table 2).
Table 2 — Target analytes of this European Standard
Target analyte a
CAS-RN
PCB28 2,4,4'-trichlorobiphenyl 7012–37–5
PCB52 2,2',5,5'-tetrachlorobiphenyl 35693–99–3
PCB101 2,2',4,5,5'-pentachlorobiphenyl 37680–73–2
PCB118 2,3',4,4',5-pentachlorobiphenyl 31508–00–6
PCB138 2,2',3,4,4',5'-hexachlorobiphenyl 35065–28–2
PCB153 2,2',4,4',5,5'-hexachlorobiphenyl 35065–27–1
PCB180 2,2',3,4,4',5,5'-heptachlorobiphenyl 35065–29–3
a
CAS-RN Chemical Abstracts Service Registry Number.
The limit of detection depends on the determinants, the equipment used, the quality of chemicals used
for the extraction of the sample and the clean-up of the extract.
Under the conditions specified in this European Standard, limit of application of 1 µg/kg (expressed as
dry matter) can be achieved.
Sludge and treated biowaste may differ in properties and also in the expected contamination levels of
PCBs and presence of interfering substances. These differences make it impossible to describe one
general procedure. This European Standard contains decision tables based on the properties of the
sample and the extraction and clean-up procedure to be used.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
EN 15934, Sludge, treated biowaste, soil and waste — Calculation of dry matter fraction after
determination of dry residue or water content
EN 16179, Sludge, treated biowaste and soil — Guidance for sample pretreatment
EN ISO 5667-15, Water quality — Sampling — Part 15: Guidance on the preservation and handling of
sludge and sediment samples (ISO 5667-15)
EN ISO 16720, Soil quality — Pretreatment of samples by freeze-drying for subsequent analysis
(ISO 16720)
EN ISO 22892, Soil quality — Guidelines for the identification of target compounds by gas
chromatography and mass spectrometry (ISO 22892)
ISO 8466-1, Water quality — Calibration and evaluation of analytical methods and estimation of
performance characteristics — Part 1: Statistical evaluation of the linear calibration function
5
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SIST EN 16167:2018
EN 16167:2018 (E)
ISO 18512, Soil quality — Guidance on long and short term storage of soil samples
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
• IEC Electropedia: available at http://www.electropedia.org/
• ISO Online browsing platform: available at http://www.iso.org/obp
3.1
polychlorinated biphenyl
PCB
biphenyl substituted by one to ten chlorine atoms
[SOURCE: EN 15308:2016, 3.1]
3.2
congener
member of the same kind, class or group of chemicals, e.g. anyone of the two hundred and nine
individual PCB
Note 1 to entry: The IUPAC congener numbers are for easy identification; they do not represent the order of
chromatographic elution.
[SOURCE: EN 15308:2016, 3.2]
3.3
critical pair
pair of congeners that will be separated to a predefined degree (e.g. R = 0,5) to ensure chromatographic
separation meets minimum quality criteria
[SOURCE: EN 15308:2016, 3.6]
3.4
internal standard
13
C -labelled PCB or other PCB that are unlikely to be present in samples added to the sample before
12
extraction and used for quantification of PCB content
[SOURCE: EN 15308:2016, 3.4, modified – "waste samples" is replaced here with "samples".]
3.5
injection standard
13
C -labelled PCB or other PCB that is unlikely to be present in samples added to the sample extract
12
before injection into the gas chromatograph, to monitor variability of instrument response and the
recovery of the internal standards
[SOURCE: EN 15308:2016, 3.5, modified – "waste samples" is replaced here with "samples".]
6
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SIST EN 16167:2018
EN 16167:2018 (E)
4 Principle
Due to the horizontal character of this European Standard, different procedures for different steps
(modules) are allowed. Which modules should be used depends on the sample. A recommendation is
given in this European Standard. Performance criteria are described and it is the responsibility of the
laboratories applying this European Standard to show that these criteria are met. Using of spiking
standards (internal standards) allows an overall check on the efficiency of a specific combination of
modules for a specific sample. But it does not necessarily give the information upon the extensive
extraction efficiency of the native PCB bonded to the matrix.
After pretreatment according to the methods referred to in 9.2, the test sample is extracted with a
suitable solvent.
The extract is concentrated by evaporation: If necessary, interfering compounds are removed by a
clean-up method suitable for the specific matrix. The eluate is concentrated by evaporation.
The extract is analysed by gas chromatography with either mass spectrometric (GC-MS) or electron
capture detection (GC-ECD). Applying GC-MS the various compounds are separated using a capillary
column with a stationary phase of low polarity. In case of GC-ECD, extracts are analysed using two
columns of different polarity (see 8.2.1).
PCBs are identified and quantified by comparison of relative retention times and relative peak heights
(or peak areas) with respect to internal standards added. The efficiency of the procedure depends on
the composition of the matrix that is investigated.
5 Interferences
5.1 Interference with sampling and extraction
Use sampling containers of materials (preferably of steel, aluminium or glass) that do not change the
sample during the contact time. Avoid plastics and other organic materials during sampling, sample
storage or extraction. Keep the samples from direct sunlight and prolonged exposure to light.
During storage of the samples, losses of PCBs may occur due to adsorption on the walls of the
containers. The extent of the losses depends on the storage time.
5.2 Interference with GC
Substances that co-elute with the target PCB may interfere with the determination. These interferences
may lead to incompletely resolved signals and may, depending on their magnitude, affect accuracy and
precision of the analytical results. Peak overlap does not allow an interpretation of the result.
Asymmetric peaks and peaks being broader than the corresponding peaks of the reference substance
suggest interferences.
Chromatographic separation between the following pairs can be critical. The critical pair PCB28 and
PCB31 is used for selection of the capillary column (see 8.2.2). If molecular mass differences are
present, quantification can be made by mass selective detection. If not or using ECD, the specific PCB is
reported as the sum of all PCBs present in the peak. Typically, the concentrations of the co-eluting
congeners compared to those of the target congeners are low. When incomplete resolution is
encountered, peak integration shall be checked and, when necessary, corrected.
— PCB28 – PCB31
— PCB52 – PCB73
— PCB101 – PCB89 / PCB90
7
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SIST EN 16167:2018
EN 16167:2018 (E)
— PCB118 – PCB106
— PCB138 – PCB164 / PCB163
Presence of considerable amounts of mineral oil in the sample may interfere with the quantification of
PCB in GC-MS. In presence of mineral oil, GC-ECD may be preferred or mineral oil can be removed using
clean-up procedure G (see 10.4.8) using DMF/n-hexane.
Presence of tetrachlorobenzyltoluene (TCBT)-mixtures may disturb the determination of the PCB with
GC-ECD.
6 Safety remarks
PCBs are highly toxic and shall be handled with extreme care. Contact between the body and solid
materials, solvent extracts and solutions of standard PCB shall not be allowed to occur. It is strongly
advised that standard solutions are prepared centrally in suitably equipped laboratories or are
purchased from suppliers specialized in their preparation.
Solvent solutions containing PCB shall be disposed of in a manner approved for disposal of toxic wastes.
For the handling of hexane precautions shall be taken because of its neurotoxic properties.
National regulations shall be followed with respect to all hazards associated with this method.
7 Reagents
All reagents shall be of recognized analytical grade. The purity of the reagents used shall be checked by
running a blank test as described in 10.1. The blank shall be less than 50 % of the lowest reporting limit.
7.1 Reagents for extraction
7.1.1 Propanone (Acetone), (CH ) CO.
3 2
7.1.2 n-heptane, C H .
7 16
7.1.3 Petroleum ether, boiling range 40 °C to 60 °C.
Hexane-like solvents with a boiling range between 40 °C and 89 °C are allowed.
7.1.4 Anhydrous sodium sulfate, Na SO :
2 4
The anhydrous sodium sulfate shall be kept carefully sealed.
7.1.5 Distilled water or water of equivalent quality, H O.
2
7.1.6 Sodium chloride, NaCl, anhydrous.
7.1.7 Keeper substance. High boiling compound, i.e. octane, nonane.
7.2 Reagents for clean-up:
7.2.1 Clean-up A using aluminium oxide:
7.2.1.1 Aluminium oxide, Al O .
2 3
2
Basic or neutral, specific surface 200 m /g, activity Super I according to Brockmann.
8
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SIST EN 16167:2018
EN 16167:2018 (E)
7.2.1.2 Deactivated aluminium oxide
Deactivated with approximately 10 % water.
Add approximately 10 g of water (7.1.5) to 90 g of aluminium oxide (7.2.1.1). Shake until all lumps have
disappeared. Allow the aluminium oxide to condition before use for some 16 h, sealed from the air, use
it for maximum two weeks.
NOTE The activity depends on the water content. It can be necessary to adjust the water content.
7.2.2 Clean-up B using silica gel 60 for column chromatography
7.2.2.1 Silica gel 60, particle size 63 µm to 200 µm.
7.2.2.2 Silica gel 60, water content: mass fraction w(H O) = 10 %.
2
Silica gel 60 (7.2.2.1), heated for at least 3 h at 450 °C, cooled down and stored in a desiccator
containing magnesium perchlorate or a suitable drying agent. Before use heat at least for 5 h at 130 °C
in a drying oven. Then allow cooling in a desiccator and add 10 % water (mass fraction) in a flask. Shake
for 5 min intensively by hand until all lumps have disappeared and then for 2 h in a shaking device.
Store the deactivated silica gel in the absence of air, use it for maximum of two weeks.
7.2.3 Clean-up C using gel permeation chromatography (GPC)
® 1)
7.2.3.1 Bio-Beads S-X3 .
7.2.3.2 Ethyl acetate, C H O .
4 8 2
7.2.3.3 Cyclohexane, C H .
6 12
®
Preparation of GPC, for example: Put 50 g Bio-Beads S-X3 (7.2.3.1) into a 500 ml Erlenmeyer flask and
add 300 ml elution mixture made up of cyclohexane (7.2.3.3) and ethyl acetate (7.2.3.2) 1:1 (volume) in
order to allow the beads to swell; after swirling for a short time until no lumps are left, maintain the
flask closed for 24 h. Drain the slurry into the chromatography tube for GPC. After approximately
three days, push in the plungers of the column so that a filling level of approximately 35 cm is obtained.
To further compress the gel, pump approximately 2 l of elution mixture through the column at a flow
−1
rate of 5 ml · min and push in the plungers to obtain a filling level of approximately 33 cm.
®2)
7.2.4 Clean-up D using Florisil .
®
7.2.4.1 Florisil , baked 2 h at 600 °C. Particle size 150 µm to 750 µm.
7.2.4.2 Iso-octane, C H .
8 18
®
1) Bio-Beads is an example of a suitable product available commercially. This information is given for the
convenience of users of this document and does not constitute an endorsement by CEN of this product. Equivalent
products may be used if they can be shown to lead to the same results.
®
2) Florisil is a trade name for a prepared diatomaceous substance, mainly consisting of anhydrous magnesium
silicate. This information is given for the convenience of users of this European Standard and does not constitute
an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same
results.
9
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SIST EN 16167:2018
EN 16167:2018 (E)
7.2.4.3 Toluene, C H .
7 8
7.2.4.4 Iso-octane/Toluene 95/5 (v/v).
7.2.5 Clean-up E using silica H SO /silica NaOH.
2 4
7.2.5.1 Silica, SiO , particle size 70 µm to 230 µm, baked at 180 °C for a minimum of 1 h, and stored
2
in a pre-cleaned glass bottle with screw cap that prevents moisture from entering.
7.2.5.2 Silica, treated with sulfuric acid.
Mix 56 g silica (7.2.5.1) and 44 g sulfuric acid (7.2.8.1).
7.2.5.3 Sodium hydroxide solution, c(NaOH) = 1 mol/l.
7.2.5.4 Silica, treated with sodium hydroxide.
Mix 33 g silica (7.2.5.1) and 17 g sodium hydroxide (7.2.5.3).
7.2.5.5 n-hexane, C H .
6 14
7.2.6 Clean-up F using benzenesulfonic acid/sulfuric acid
7.2.6.1 3 ml silica gel column, of adsorbent mass 500 mg, particle size 40 µm.
7.2.6.2 3 ml benzenesulfonic acid/sulfuric acid column, of adsorbent mass 500 mg, particle size
40 µm.
7.2.7 Clean-up G using DMF/hexane partitioning
7.2.7.1 Dimethylformamide(DMF), C H NO.
3 7
7.2.8 Clean-up H using concentrated sulfuric acid
7.2.8.1 Sulfuric acid, H SO of purity 96 % to 98 % (mass fraction).
2 4
7.2.9 Clean-up I using TBA sulfite reagent
7.2.9.1 Tetrabutylammonium reagent (TBA sulfite reagent)
Saturate a solution of tetrabutylammonium hydrogen sulfate in a mixture of equal volume of water and
2-propanol, c((C H ) NHSO ) = 0,1 mol/l, with sodium sulfite.
4 9 4 4
25 g of sodium sulfite should be sufficient for 100 ml of solution.
7.2.9.2 2-Propanol, C H O.
3 8
7.2.9.3 Sodium sulfite, Na SO .
2 3
7.2.10 Clean-up J using pyrogenic copper
WARNING — Pyrogenic copper is spontaneously inflammable. Suitable precautions shall be taken.
7.2.10.1 Copper(II)-sulfate pentahydrate, CuSO · 5 H O.
4 2
10
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SIST EN 16167:2018
EN 16167:2018 (E)
7.2.10.2 Hydrochloric acid, c(HCl) = 2 mol/l.
7.2.10.3 Zinc granules, Zn, particle size 0,3 mm to 1,4 mm.
7.2.10.4 Anionic detergent aqueous solution (e.g. 35 g/100 ml, n-dodecane-1-sulfonic acid sodium
salt (CH (CH ) SO Na).
3 2 11 3
Other commercially available detergents may also be suitable.
7.2.10.5 Deoxygenated water
Water from which dissolved O has been removed, e.g. by stripping with an inert gas, or by adequate
2
membrane filtration.
7.2.10.6 Pyrogenic copper
Dissolve 45 g copper(II)-sulphate pentahydrate (7.2.10.1) in 480 ml water containing 20 ml
hydrochloric acid (7.2.10.2) in a 1 000 ml beaker.
Take 15 g of zinc granules size (7.2.10.3), add 25 ml water and one drop of anionic detergent solution
(7.2.10.4) in another 1 000 ml beaker.
Stir with a magnetic stirrer at a high speed to form a slurry. Then whilst stirring at this high speed,
carefully add the copper(II)-sulphate solution drop by drop using a glass rod.
Hydrogen is liberated and elemental pyrogenic copper is precipitated (red coloured precipitate).
Stirring is continued until the hydrogen generation almost ceases. Then the precipitated copper is
allowed to settle. The supernatant water is carefully removed and the product washed with
deoxygenated water (7.2.10.5) three times, to eliminate residual salts.
Then the water is carefully replaced with 250 ml acetone (7.1.1) (whilst continuously stirring the
mixture). This operation is repeated twice more to ensure elimination of water.
Then the above procedure is repeated three times with 250 ml hexane (7.2.5.5), to ensure elimination
of the acetone.
Carefully transfer the copper with hexane into an Erlenmeyer flask and store under hexane. The flask
shall be sealed to prevent ingress of air and stored in an explosion-proof refrigerator 2 °C to 8 °C.
The shelf life of the pyrogenic co
...
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