HPLC Column c18 With Mobile Phase For Succinylsulfathizaole PG 18

High performance HPLC packings
and
Applications
Nucleodur int2/5/0/10.2005 BD
Printed in Germany
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MACHEREY-NAGEL
DIN EN ISO 9001: 2000

CERTIFIED
MACHEREY-NAGEL
MN
NUCLEODUR Reversed Phase Packings

In RP liquid chromatography the efficiency of the packing is strongly affected by the quality of the base
silica itself. Shortcomings in the surface geometry of the particles or metal contaminants are the main
reasons for inadequate coverage with the covalently bonded alkylsilanes in the subsequent derivatization
steps. It is well known, that poor surface coverage and, in consequence, high activity of residual free
silanols often results in peak tailing or adsorption, particularly with basic compounds.
NUCLEODUR Particle Shape and Surface Symmetry

Billiard Balls in HPLC
NUCLEODUR silicas are synthesized in a unique and carefully
controlled manufacturing process which provides silica particles, which are totally spherical. The pictures show the uniform particle size distribution and the outstanding smoothness of the NUCLEODUR surface is emphasized.
Fig. 1: High performance totally spherical NUCLEODUR silica
Fig. 2: Surface of a NUCLEODUR particle (magnified)
NUCLEODUR Purity Intolerant Towards Metals

As already mentioned above, a highly pure silica is required for
achieving symmetric peak shapes and maximum resolution.
Inclusions of e.g. iron or alkaline earth
metal ions on the silica surface are
Aluminium
<5
ppm
largely responsible for the unwanted
Iron
<5
ppm
<5
ppm
interactions with ionizable analytes, e.g. Sodium
Calcium
< 10
ppm
amines or phenolic compounds.
Titanium
<1
ppm
NUCLEODUR is virtually free of metal
Zirconium
<1
ppm
impurities and low acidic surface
Arsenic
< 0.5
ppm
silanols. Elemental analysis data of
Mercury
< 0.05 ppm
NUCLEODUR 5 m measured by AAS

Elementary analysis (metal ions)
are listed beside.
of NUCLEODUR 100-5
NUCLEODUR Reversed Phase Packings

NUCLEODUR Pressure Stability Hard as a Rock
The totally spherical and 100% synthetic silica gel exhibits an outstanding
mechanical stability, even at high pressures and elevated eluent flow rates.
In addition, after several cycles of repeated packing, no significant drop in
pressure can be observed. The latter is of prime importance for preparative
and process-scale applications.
120800
2
3
250 inj.
200 inj.
150 inj.
50 inj.
1
0
Column:
Eluent:
Temperature:
Detection:
start
1
min 4
125 x 4 mm NUCLEODUR 100-5 C18 ec

acetonitrile water (80:20, v/v)
25 C
UV, 254 nm
Peaks:
1. Phenol
2. Naphthalene
3. Anthracene
Fig. 3: Column stability of NUCLEODUR 100-5 C18 ec
back pressure [bar]
100 inj.
120
80
40
Packing #
Fig. 4: Repeated packing
NUCLEODUR C18 ec
the Workhorse for
Daily Routine Analysis
and Up-Scaling for Preparative HPLC
The efficiency of a separation is controlled by particle size and selectivity of the stationary phase.
The exceptional surface coverage of monomeric
bonded alkylsilanes, combined with an exhaustive
endcapping, results in a surface with lowest silanol
activity. This allows the tailing-free elution of polar
compounds such as basic drugs. NUCLEODUR
C18 ec is available in 9 different particle sizes (3, 5,
7, 10, 12, 16, 20, 30 and 50 m) which cover the
whole range from high speed analytical HPLC up to
medium and low pressure prep LC. NUCLEODUR
C18 ec is also an ideal tool for scale-up purposes.
Surface area (BET)

Pore size
Pore volume
Carbon load
340 m2/g
110
0.9 ml/g
17.5 %
Physical data of NUCLEODUR C18 ec
NUCLEODUR C18 Gravity

The column for
method development
HPLC at pH extremes
overall sophisticated analytical separations
NUCLEODUR C18 Gravity is based on the ultrapure NUCLEODUR silica, which is described
above.
An unique derivatization process generates a homogeneous surface with a high density of bonded
silanes (carbon content ~ 18%). The following thorough endcapping suppresses any unwanted polar
interactions between the silica surface and the
sample, which makes Gravity particularly suitable for the separation of basic and other ionizable
analytes. Figure 5 shows a comparison study,
where the strongly basic amitriptyline is eluted on
various highly base deactivated C18 phases under
isocratic conditions.
More examples to emphasize the outstanding suitability of the NUCLEODUR C18 Gravity phase are
shown below (figures 6 and 7) and in the application section of this brochure.
120810
Base deactivation
Phase S (C18, 5 m)
Phase P (C18, 5 m)
Phase L (C18, 5 m)
1 and 2 overlap
3
1
Phase I (C18, 5 m)
2
NUCLEODUR C18 Gravity, 5 m
10
15
20
25 min
Columns:
Eluent:
250 x 4 mm
methanol 20 mM KH2PO4,
pH 7.0 (75 : 25, v/v)
Flow rate:
1.0 ml/min
Temperature: 30 C
Detection:
UV, 254 nm
Peaks:
1. Dibutyl
phthalate
2. Acenaphthene
3. Amitriptyline
Peaks:
1. Pyridine
2. Phenol
120830
120820
Fig. 5: Comparison of different base deactivated phases
6
10
4 5
2
0
10
15
20 min
Column: 250 x 4 mm NUCLEODUR C18 Gravity, 5 m

Eluent: methanol 20 mM KH2PO4, pH 7.0 (75 : 25, v/v)
Flow rate:
1.0 ml/min
Detection:
UV, 254 nm
Temperature: 30 C
Inj. volume:
5 l
0
Column:
Eluent:
Flow rate:
Temperature:
10
250 x 4 mm NUCLEODUR C18 Gravity, 5 m

methanol water (30 : 70, v/v)
1.0 ml/min
Detection:
UV, 254 nm
40 C
Inj. volume:
5 l
Fig. 6: Separation of pyridine and phenol
min
Peaks:
1. Uracil
2. 2,7-Dihydroxynaphthalene
4. Lidocaine
5. Toluene
Fig. 7: Selectivity test
6. Naphthalene
7. Ethylbenzene
8. Dibutyl phthalate
9. Acenaphthene
10. Amitriptyline

Enhanced pH Stability
One major disadvantage of using silica stationary
phases is the limited stability at strongly acidic or
basic pH ranges. Cleavage of the siloxane bonding
by hydrolysis, or dissolution of the silica will rapidly lead to a considerable loss in column performance. Therefore conventional RP phases are usually not recommended to be run with mobile
phases at pH > 8 or pH < 2 for extended periods of
time. The special surface bonding technology and
the low concentration of trace elements of
NUCLEODUR C18 Gravity allow for use at an expanded pH range from pH 1 to 11.
Figure 8 shows the extent of protonation of surface

silanols and of two exemplary analytes at acidic
and alkaline pH. The graph in figure 9 explains the
general correlation between retention and pH.
B
HA
k
When is enhanced pH stability beneficial?

A
BH+
0
12
pH
Fig. 9: Correlation between retention and pH for basic and acidic
compounds
In figure 10 the excellent column stability of

NUCLEODUR C18 Gravity in acidic conditions is
shown. The retention time of all three compounds
in the column performance test remains consistent
and virtually unchanged, even after the column is
run with 5000 ml eluent. Due to the extremely stable
surface modification, no cleavage of the Si-O-Si
bonding occurs, column deterioration is therefore
successfully prevented.
120840
The option to work at an expanded pH range is often required in method development. Many nitrogen containing compounds like basic drugs are
protonated at acidic or neutral pH and exhibit poor
retention on a standard C18 phase. The retention
behavior can be improved by working at a higher
pH, where the analyte is no longer protonated, but
formally neutrally charged, as a rule between pH 9
10. For acidic analytes it is exactly in inverse proportion, maximum retention can be attained at low
pH.
2
3
after 5000 ml eluent
1st injection
10 min
Column: 125 x 4 mm NUCLEODUR C18 Gravity, 5 m

eluent: acetonitrile 1% TFA in water (50:50, v/v), pH 1.5
flow rate: 1.0 ml/min, temperature: 30 C,
detection: UV, 230 nm, inj.volume: 5 l
Peaks: 1. pyridine, 2. toluene, 3. ethylbenzene
Fig. 8: Surface silanols at different pH values
Fig. 10: Stability of NUCLEODUR C18 Gravity at pH 1.5

Fig. 11: Stability of NUCLEODUR C18
Gravity under alkaline conditions compared with different C18 phases
first injection
after 300 injections
Phase X
Phase L
2 min
Columns:
Eluent:
2
3 min
50 x 4.6 mm
methanol water ammonia
(20:80:0.5, v/v/v), pH 11
Flow rate:
1.3 ml/min
Temperature: 30 C
Detection:
UV, 254 nm
Inj. volume:
2.0 l
Peaks:
1. Theophylline
2. Caffeine
NUCLEODUR
C18 Gravity
2 min
Phase I
4 min
Fig. 11 demonstrates the stability of NUCLEODUR

C18 Gravity under alkaline conditions in comparison with 4 commercially available modern RP18
phases. Again, the ultrapure Gravity with its unique
high density surface bonding technology withstands strong alkaline mobile phase conditions.
Even after 300 injections no loss of column efficiency, identified e.g. by
peak broadening or decrease
in retention times,
could be observed.
3 min
120850
Phase K
As it was
previously mentioned, pH stability of
the stationary phase can be
helpful for improving selectivity in
method development. Figure 12 shows
the separation of 4 basic drugs under acidic
and basic conditions.
118010
2
10 min
5
2
A)
3
1
120860
pH 2.5
The pH stability of silica under alkaline conditions

is mainly a kinetic effect and based on the velocity
of the dissolution of the silica support. It is worth
mentioning, that this phenomenon also depends
on type and concentration of buffers, as well as on
the temperature. It is well known, that the use of
phosphate buffers, particularly at elevated temperatures, can reduce column lifetime even at moderate pH. If possible, phosphate buffers should be replaced by less harmful alternatives.
1
4
pH 10.0
3
0
10 min
Column:
Eluent A:
acetonitrile
Eluent B:
20 mM (NH4)2HPO4, pH 2.5 / 10.0
Gradient:
10% A (1 min) 75% A in 10 min
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj. volume:
2 l
Peaks:
1. Lidocaine
2. Papaverine
3. Noscapine
4. Diphenhydramine
Fig. 12: Separation of basic alkaloids
At pH 2.5 the protonated analytes exhibit poor retention (early elution) and in addition an inadequate
resolution for papaverine and noscapine, whilst the
formally non ionized molecules can be baseline
separated due to the better retention pattern at alkaline pH.
A further example how selectivity can be controlled
by the pH value is demonstrated in figure 13. The
sample mixture consists of an acid (ketoprofen), a
base (lidocaine) and benzamide. Under acidic conditions the protonated lidocaine is eluted very fast
due to lack of sufficiently strong hydrophobic interactions between analyte and C18 chains, in contrary to the formally neutral ketoprofen, which is
eluted after about 3 minutes. However at pH 10 a reversal of the elution order, with a visibly longer retention time for the basic lidocaine, can be
achieved.
pH 3.0
4 min
B)
3
1
pH 10.0
4 min

A) acetonitrile 10 mM ammonium formate,
pH 3.0 (50:50, v/v)
B) acetonitrile 10 mM ammonium bicarbonate,
pH 10.0 (50:50, v/v)
Flow rate:
1.0 ml/min
Temperature: 30 C
Detection:
UV, 230 nm
Inj. volume:
2 l
Peaks:
1. Lidocaine
2. Benzamide
3. Ketoprofen
Column:
Eluents:
CH3
NH
NH2
N(C2H5)2
H3C O
OH
H3C
O
Fig. 13: Influence of the pH value on selectivity
NUCLEODUR C18 Pyramid

RP-HPLC with Highly Aqueous Mobile Phases
stable in 100% aqueous mobile phase systems
interesting polar selectivity features
excellent base deactivation
pH stability 2 8
The efforts to neutralize unwanted activity of unreacted surface silanols often results in well basedeactivated phases with high carbon load, but a
limited scope of selectivity beyond non-polar interactions. In particular polar compounds like carboxylic acids, drug metabolites, etc. show only weak
retention on densely bonded reversed phase columns due to distinct hydrophobic properties but
low polar selectivity. Very polar analytes require
highly aqueous mobile phases for solubility and retention. Conventional reversed phase columns often display stability problems in eluent systems
with high percentage of water (> 95%) as evidenced
by a sudden decrease of retention time and overall
poor reproducibility. This phenomenon is described as phase collapse caused by the mobile
phase expelled from the pores due to the fact,
that hydrophobic RP phases are incompletely wetted with the mobile phase 1).
Different approaches can be used
to increase column stability with
highly aqueous mobile phase
systems. The most promising
concepts are incorporating a
polar group in the hydrophobic alkyl chain, or using hydrophilic endcapping procedures to improve the
wettability of the reversed
phase modification.
NUCLEOSIL NAUTILUS
may be taken as an example
for the embedded polar
group strategy, in which a
C18 silane with a polar function is successfully linked to
the silica surface 2).
1) U. D. Neue et al., Chromatographia 54 (2001) 169 177

2) D. Rieger, H. Riering, Int. Laboratory Aug. 2000,
Vol. 30 (4A), 12
3) D. Rieger, J. Pfeiffer, LaborPraxis 26(2) (2002) 20 21
Stability features
NUCLEODUR C18 Pyramid is a silica phase with
hydrophilic endcapping, designed especially for
use in eluent systems of up to 100% water. In figure
16 we studied the retention behaviour of tartaric,
acetic and maleic acid under purely aqueous conditions on NUCLEODUR C18 Pyramid in comparison with a conventionally bonded RP phase.
It can be shown that the retention times for
NUCLEODUR C18 Pyramid remain nearly unchanged between initial injection and restart after
the flow has been stopped for 12 hours (see figure
16), whilst the performance of the conventional RP
column collapsed totally after the same period.

Retention characteristics
119170
Based on the ultrapure NUCLEODUR silica 3) the

polar surface derivatization exhibits retention characteristics, which differentiate the Pyramid from
conventional C18 stationary phases. Figure 14
shows the improved retention behavior of very polar compounds such as short chain organic acids,
which are insufficiently retained on RP columns
with predominantly hydrophobic surface properties.
Figure 15 shows the separation of various organic

acids at acidic pH under 100% aqueous eluent conditions.
1
3
2
10 min
NUCLEODUR
Column:
C18
Pyramid, 5 m, 250 x 4 mm
Eluent:
0.2% H3PO4
Flow rate:
0.7 ml/min
Temperature: 25 C
Detection:
UV, 210 nm
Peaks:
(injection volume 2 l)
1. Tartaric acid
2. Malic acid
3. Lactic acid
4. Succinic acid
Fig. 15: Separation of organic acids
120870
119180
t0
Column: NUCLEODUR C18

Pyramid, 5 m, 125 x 4 mm
Eluent:
0.2% H3PO4
Flow rate:
1.0 ml/min
Temperature: 22 C
Detection:
UV, 202 nm
Peaks:
(injection volume 2 l)
1. Formic acid
2. Acetic acid
2
4 min
0
Fig. 14: Separation of very polar compounds
2
conventional RP column
3
3
initial
injection
1
2
5 min
5 min
pump
stopped!
Both columns:
Inj. volume:
Eluent:
Flow rate:
Temperature:
Detection:
restart
after 12 h
5 min
125 x 4 mm
1 l
50 mM KH2PO4 pH 2.5
0.7 ml/min
25 C
UV, 210 nm
Peaks:
1. Tartaric acid
2. Acetic acid
3. Maleic acid
0
5 min
Fig. 16: Stability test comparison

Base deactivation
In addition to the exceptional polar selectivity

NUCLEODUR C18 Pyramid also provides adequate hydrophobic retention, which can be illustrated in the selectivity test according to Engelhardt 4) (see figure 17). The capacity factors of the
non-polar, alkyl-substituted benzenes toluene and
ethylbenzene do not go too far in comparison with
standard C18 phases.
Peaks:
1. Uracil
2. Aniline
3. Phenol
4. Dimethylaniline
5. p-Ethylaniline
6. Toluene
7. Ethylbenzene
1
2
4
6
The perceptible increase in polarity has no impact

on the retention behavior of ionizable analytes.
Even with the strongly basic compounds of the tricyclic antidepressant drug test mixture, no unwanted interactions or a so-called lack in base deactivation are observed (see figure 18).
Peaks:
1. Protriptyline
2. Nortriptyline
3. Doxepin
4. Imipramine
5. Amitriptyline
3
4
2
1
5
5
7
10
20
30 min
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
119190
250 x 4 mm NUCLEODUR C18 Pyramid, 5 m

methanol water (55:45, v/v)
1.0 ml/min
40 C
UV, 254 nm
3 l
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
Fig. 17: Selectivity test

3 4
7
Fig. 18: Tricyclic antidepressants

4
tR hydrophobic compounds
5
1
4
tR hydrophobic compounds
12
5
10 min
MeOH 20 mM NH4H2PO4 pH 6.95 (70:30, v/v)
1.0 ml/min
40 C
UV, 254 nm
5 l
119200
3 5
2
1
10
Columns:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
20
119221
tR polar compounds
tR polar compounds
30 min
250 x 4 mm, 5 m particles

methanol 25 mM NH4H2PO4, pH 7 (65:35, v/v)
0.8 ml/min
40 C
UV, 254 nm
5 l
10
20
30 min
Peaks:
1. Chlorpheniramine
2. Dimethyl phthalate
3. Benzamide
4. Ethyl benzoate
5. Benzophenone
6. Lidocaine
Fig. 19: Retention behavior of polar and non-polar compounds on different NUCLEODUR RP columns
10
6
9
119210
119220
10
15 min
7. Naphthalene
8. Biphenyl
9. Acenaphthene
NUCLEODUR C8 Phases
NUCLEODUR C8 ec and NUCLEODUR C8 Gravity
C18 or C8 the best of both worlds

Chromatographers now might wonder about the
differences between C8 and C18 phases and the
preferred range of application.
NUCLEODUR 100-5 C8 ec
1
5
Peaks:
1. Resorcinol
2. Pyrocatechol
3. Phenol
4. 4-Methoxyphenol
8
4
120890
6
1
5
5. 2-Methoxyphenol
6. 2-Ethoxyphenol
7. Veratrol
8. Biphenyl-2-ol
9. Phenetole
2
4
8 9
120891
In addition to the program of NUCLEODUR C18

phases MACHEREY-NAGEL offers the corresponding octyl modified NUCLEODUR C8 ec and
NUCLEODUR C8 Gravity columns to expand the
reversed phase tool box effectively. Based on the
same totally spherical and highly pure silica the C8
phases exhibit the same excellent chemical and
mechanical stability features as the C18 counterparts. Indeed NUCLEODUR C8 Gravity can also be
run at pH extremes (pH 1 11) by choosing appropriate elution parameters. Due to the shorter chain
and less hydrophobic properties of the stationary
phase the retention of non-polar compounds is decreased, and in consequence a reduction in time of
analysis can be achieved. Moreover a stronger polar selectivity, particularly with the separation of
ionizable analytes is frequently observed (as distinct from the C18 phases). NUCLEODUR C8 ec
and NUCLEODUR C8 Gravity are most suitable for
the development of new methods but also for robust routine analysis.

0
Columns:
Flow rate:
Temperature:
Detection:
Inj. volume:
5
250 x 4 mm
1.0 ml/min
25 C
UV, 275 nm
10 l
10
min
15
20
Eluents: A) water, B) methanol

Gradient for C8: 2 min 20% B,
then to 60% B in 12 min
Gradient for C18: 2 min 25% B,
then to 65% B in 12 min
Fig. 21: Phenols

Peaks:
1. Piroxicam
2. Suprofen
3. Ketoprofen
4. Carprofen
5. Fenoprofen
6. Diclofenac
4+5
3
6
120880
C8
1+2
4 5
120881
C18
0
10
15
250 x 4 mm NUCLEODUR 100-5 C
min
20
Column:
8 ec / C18 ec
Eluent:
acetonitrile water, 1% acetic acid (48 : 52, v/v)
Flow rate:
1.0 ml/min
Detection:
UV, 230 nm
Temperature: 25 C
Inj. volume:
10 l
Fig. 20: Anti-inflammatory drugs
Indeed there are no general guidelines which could

make the choice easier but it will always be beneficial to add both phases to the existing pool of reversed phase columns in the laboratory.
The two comparative studies reveal some different
selectivity patterns of NUCLEODUR C8 ec and
NUCLEODUR C18 ec. In figure 21 baseline separation for 2-ethoxyphenol and dimethoxybenzene
(veratrol) and in addition a reversal of the elution
order of phenol and 4-methoxyphenol can be
shown on the octyl phase. The separation of various nonsteroidal anti-inflammatory drugs (figure
20) illustrates the differences in polarity between
C8 and C18 and the resulting impact on efficiency.
NUCLEODUR C8 ec exhibits enhanced selectivity
and excellent resolution for the polar compounds
piroxicam and suprofen which co-elute on the C18
column. However due to the longer alkyl chain
NUCLEODUR C18 ec shows a distinct hydrophobic selectivity that leads to baseline separation of
the more non-polar analytes carprofen and fenoprofen with superior peak shapes.
11
NUCLEODUR Sphinx RP
The Bifunctional RP Phase
distinct selectivity based on bifunctional surface coverage
widens the scope in method development
high density of covalently bonded silanes
guarantees sharp peaks without tailing
pH stabilility can be used from pH 1 10
low bleeding characteristics in LC/MS applications
high reproducibility tight QC procedures provide consistant quality
NUCLEODUR Sphinx RP is characterized by exceptional selectivity features generated by a wellbalanced ratio of covalently bonded octadecyl and
phenyl groups. The combination of classical hydro interactions (aromatic ring system)
phobic with -
expands the scope of selectivity in comparison
with conventional reversed phase packings. NUCLEODUR Sphinx RP is particularly suited for the
separation of molecules containing aromatic and
multiple bonds. For the separation of polar compounds NUCLEODUR Sphinx RP can be especially recommended and can also outperform many
customary C18 phases. In addition, exhaustive endcapping steps minimize unwanted surface silanol
activity and guarantee excellent peak shapes even
for strong basic analytes.
Different from standard phenyl phases,

NUCLEODUR Sphinx RP is far more stable towards hydrolysis and is also suggested for LC/MS
applications.
Peaks:
1. Theophyllline
2. Caffeine
1
2
after 300 injections

(column run
with 5 l eluent)
1st injection
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
min
50 x 4,6 mm NUCLEODUR Sphinx RP, 5 m

methanol dil. NH3, pH 10 (20:80, v/v)
1.0 ml/min
30 C
UV, 275 nm
3 l
Fig. 22: Stability at pH 10
120900
Due to the additional intermolecular interactions

NUCLEODUR Sphinx RP is an interesting replenishment to the high density bonded phases
NUCLEODUR C8/C18 Gravity and the polar
endcapped NUCLEODUR C18 Pyramid.
12
NUCLEODUR Sphinx RP
The Bifunctional RP Phase
1
2
2
NUCLEODUR
Columns:
Eluent:
Flow rate:
Temperature:
6 min
Sphinx-RP, 5 m
6 min
Competitor 1 (column XP)
6 min
Competitor 2 (column LP)
6 min
Competitor 3 (column SP)
Detection:
UV, 254 nm
Peaks (inj. volume 2 l):
1. Pyridine
2. Phenol
150 x 4.6 mm
1.0 ml/min
40 C
120910
Fig. 22: Comparison of surface deactivation of different phenyl modified RP phases
Columns:
Eluent:
Flow rate:
Temperature:
Inj. volume:
Detection:
150 x 4.6 mm
water methanol (40:60, v/v)
1 ml/min
30 C
3 l
270 nm
Peaks:
1. Catechin
OH
OH
O
HO
OH
OH
R1
OH
O
HO
R2
R3
2. Rutin
3. Fisetin
4. Quercetin
5. Kaempferol
6. Isorhamnetin
R1 = R3 = OH, R2 = O-rutinose
R1 = R2 = OH, R3 = H
R1 = R2 = R3 = OH
R1 = H, R2 = R3 = OH
R1 = OCH3, R2 = R3 = OH
NUCLEODUR Sphinx RP, 5 m
0.0
3
2.5
5
5.0
6
7.5
Fig. 23: Separation of flavonoids on 3 different NUCLEODUR phases
min
10.0
119830
13
NUCLEODUR CN and CN-RP

Cyano-modified High-purity Silica
Widen your scope in selectivity!
multi-mode column (RP and NP)
different retention characteristics in comparison to C8 / C18
stable against hydrolysis at low pH, pH working range 1 8
high reproducibility from lot to lot
Peaks:
1. Benzamide
2. Dimethyl phthalate
3. Phenetole
4. o-Xylene
5. Biphenyl
5
3
lot D
lot C
lot B
lot A
min
250 x 4 mm NUCLEODUR 100-5 CN-RP

1.0 ml/min
20 C
UV, 254 nm
5 l
Fig. 24: Reproducibility of NUCLEODUR CN
14
120380
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
Peaks:
1. Maleic acid
2. Norephedrine
3. Ephedrine
4. Acetaminophen
5. Chlorpheniramine
6. Brompheniramine
3
2
0
Columns:
8
NUCLEODUR
NUCLEODUR
12 min
250 x 4 mm
100-5 C18 ec
100-5 CN-RP
250 x 4 mm
Eluent:
acetonitrile 100 mM sodium citrate pH 2.5
(15:85, v/v)
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 270 nm
Inj. volume:
10 l
Fig. 25: Comparison of NUCLEODUR C18 ec and CN-RP for

a separation of cold-medicine ingredients
119340
In reversed phase HPLC it is fairly common to start

with C18 or C8 columns, if new methods have to be
developed. However, superior polarity and selectivity properties often required for more sophisticated separations, are not always sufficiently provided by classical RP phases, which are usually
characterized by a hydrophobic layer of monomeric or polymeric bonded alkylsilanes.
One approach to improve the resolution of compounds poorly separated on nonpolar stationary
phases, is to change bonded-phase functionality.
The fully endcapped and highly reproducible (figure 24) NUCLEODUR 100-5 CN-RP phase has cyanopropyl groups on the surface able to generate a
clearly recognizable different retention behavior
compared to purely alkyl-functionalized surface
modifications (figure 25).
NUCLEODUR CN and CN-RP

Cyano-modified High-purity Silica
The polarity of the NUCLEODUR 100-5 CN-RP
phase can be classified as intermediate based on
multiple retention mechanisms such as dipole-di, and also hydrophobic interactions 1).
pole, -
Therefore, this phase shows a distinct selectivity
for polar organic compounds as well as for molecules containing -electron systems (e.g. analytes
with double bonds, tricyclic antidepressants) 2).
Short-chain bonded phases are sometimes suspected of revealing shortcomings in stability towards hydrolysis at low pH 3). The chromatogram
in figure 26 shows that even after 100 sample injections and four weeks storage at pH 1 (curve 2), neither a considerable shift in retention, nor a visible
change in peak symmetry could be noticed (curve
1 = new column).
Due to the exceptional polarity features the cyano
phase can also be run in the normal phase mode.
NUCLEODUR CN columns for normal phase applications are shipped in n-heptane. The drastic
change in selectivity and order of elution for a mixture of various steroids in normal and reversed
phase mode is displayed in figure 27 below. Moreover the high coverage combined with a thorough
endcapping makes NUCLEODUR 100-5 CN-RP
suitable for the separation of ionizable compounds
such as basic drugs.
Peaks:
1. Benzamide
2. Dimethyl
phthalate
3. Phenetole 1
4. o-Xylene
5. Biphenyl
3
5
2
1
2
4 min

acetonitrile water, 2% TFA pH 1 (50:50, v/v)
1.0 ml/min
25 C
UV, 254 nm
5 l
Fig. 26: Stability of NUCLEODUR CN-RP at pH 1
References
1) C. S. Young and R. J. Weigand, LCGC 20 (5), 464 473 (2002)
2) V. R. Meyer, Practical High Performance Liquid Chromatography (John
Wiley & Sons, New York, 3rd. ed., 1999)
3) J. J. Kirkland, LCGC 14 (6), 486 500 (1996).
2
6+7
2
3
4
10
20 min
Reversed phase mode

Column:
Eluent:
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj.volume:
10 l
Peaks :
1. Methyltestosterone
2. Testosterone
3. Norgestrel
4. Medrysone
5. Cortisone
6. Hydrocortisone
7. Prednisolone
119271
119272
1
1
119350
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
20
30 min
Normal phase mode

Column:
250 x 4 mm NUCLEODUR 100-5 CN
Eluent:
n-heptane 2-propanol (90:10, v/v)
other conditions as for reversed phase mode
Fig. 27: Separation of steroids in normal phase and reversed phase mode
15
Applications
Appl. 1: Analgesics
Appl. 3: Analgesics
Peaks:
1. Paracetamol
2. Acetylsalicylic acid
3. Salicylic acid
4. Ketoprofen
5. Diclofenac
6. Ibuprofen
Peaks:
1. Paracetamol
3. 4-Hydroxymethyl
benzoate
4. Ketoprofen
5. Flurbiprofen
6. Ibuprofen
1 2
5
4
5
6
0
0
10
12
min
min

acetonitrile 20 mM KH2PO4, pH 2.5
(50:50, v/v)
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 230 nm
Inj. volume:
5 l
Column:
Eluent:
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:

acetonitrile 0.1% TFA (50:50, v/v)
1.0 ml/min
25 C
UV, 254 nm
5 l
MN Appl. No. 117770
MN Appl. No. 119160
Appl. 2: Rapid separation of analgesics
Appl. 4: Analgesics
Peaks:
(about 0.5 g)
2. Salicylic acid
(about 0.5 g)
1
2
Peaks:
1. Paracetamol
2. Caffeine
3. 2-Acetamidophen
4. Acetanilide
6. Phenactin
A
1
3
4
B
0
0.5
min
1.5
0

(50:50, v/v)
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj. volume:
5 l
Column:
Eluent:
MN Appl. No. 117780
16
min
Column:
Eluent:
methanol 0.1% phosphoric acid (40:60, v/v)
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 240 nm
A: Thomapyrin tablet; B: standard
Thomapyrin is a trademark of Boehringer Ingelheim Pharma KG
MN Appl. No. 118600
Applications
Appl. 5: Analgesic and antiinflammatory drugs
Peaks:
1. Acetylsalicylic
acid
2. Tolmetin
3. Piroxicam
4. Suprofen
5. Naproxen
6. Diflunisal
7. Fenoprofen
8. Flurbiprofen
9. Indomethazin
10. Ibuprofen
Appl. 7: Antiinflammatory drugs
g/ml
1.6
1
26
26
26
0.64
1.6
26
26
52
52
2
6
2
9
5
1
3
7
8
8
3
10
10
0
5
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
Peaks:
1. Acetylsalicylic
acid
2. Sulindac
3. Piroxicam
4. Suprofen
5. Tolmetin
6. Naproxen
7. Diflunisal
8. Flurbiprofen
9. Indomethazin
10. Ibuprofen
10
min
15 min

acetonitrile 1% acetic acid (48:52, v/v)
1.0 ml/min
25 C
UV, 230 nm
10 l

(45:55, v/v)
Flow rate:
1.0 ml/min
Temperature: 22 C
Detection:
UV, 230 nm
Inj. volume:
5 l
Column:
Eluent:
MN Appl. No. 117840
MN Appl. No. 118590
Appl. 6: Antiinflammatory drugs
Peaks:
2. Sulindac
3. Tolmetin
4. Ketoprofen
5. Flurbiprofen
6. Diclofenac
7. Ibuprofen
8. Meclofenamic acid
2
4
Appl. 8: Barbiturates
Peaks:
1. Phenobarbital
2. Pentobarbital
3. Hexobarbital
4. Mephobarbital
5. Thiamylal
3
4
2
5
67
8
10
min
0

(45:55, v/v)
Flow rate:
1.0 ml/min
Temperature: 22 C
Detection:
UV, 230 nm
Inj. volume:
5 l
Column:
Eluent:
MN Appl. No. 117830
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
min

0.7 ml/min
25 C
UV, 254 nm
5 l
MN Appl. No. 117820
17
Applications
Appl. 9: Antidepressants
Peaks:
1. Nordoxepin
2. Protriptyline
3. Maprotiline
4. Nortriptyline
5. Norclomipramine
6. Doxepin
7. Imipramine
8. Amitriptyline
9. Clomipramine
10. Trimipramine
4
6
5
8
2
7
Appl. 11: Tricyclic antidepressants
2
4
10
10
15
Peaks:
1. Protriptyline
2. Nortriptyline
3. Doxepin
4. Imipramine
5. Amitriptyline
6. Trimipramine
min

A) methanol acetonitrile (50:50, v/v)
B) 20 mM KH2PO4, pH 7.0
50 35% B in 6 min, then 20 min at 35% B
Flow rate:
1.2 ml/min
Temperature: 30 C
Detection:
UV, 230 nm
Inj. volume:
5 l
Column:
Eluent:
Column:
Eluent:
Flow rate:
Temperature:
Detection:
10
15
min

methanol 20 mM KH2PO4, pH 7 (65:35, v/v)
1.0 ml/min
25 C
UV, 254 nm
MN Appl. No. 118520
MN Appl. No. 117790
Appl. 10: Tricyclic antidepressants
Appl. 12: Cold medicine
Peaks:
1. Protriptyline
2. Nortriptyline
3. Doxepin
4. Imipramine
5. Amitriptyline
6. Trimipramine
4
3
2
5
1
5
2
Peaks:
1. Maleic acid
2. Acetaminophen
3. Pseudoephedrine
4. Benzoic acid
5. Chlorpheniramine
6. Dextromethorphan
3
1
min

(65:35, v/v)
Flow rate:
1.0 ml/min
Temperature: 40 C
Detection:
UV, 254 nm
Inj. volume:
2 l
Column:
Eluent:
MN Appl. No. 117800
18
min

A) 50 mM KH2PO4 + 5 mM pentanesulfate
(Na salt), pH 2.5; B) methanol
Gradient:
35 55% B in 5 min
Flow rate:
1.0 ml/min
Temperature: 40 C
Detection:
UV, 230 nm
Inj. volume:
5 l
Column:
Eluents:
MN Appl. No. 117810
Applications
Appl. 13: Cold medicine ingredients
Peaks:
1. Ascorbic acid
2. Paracetamol
3. Caffeine
4. Chlorpheniramine
5. Dextromethorphan
6. Diphenhydramine
Appl. 15: Basic drugs

Peaks:
1. Procainamide
2. Clonidin
3. Clenbuterol
1
3
4
1
Column:
Eluent:
Gradient:
Flow rate:
Temperature:
Detection:
Inj. volume:
2
56
12 min
2.5
5 min
NUCLEODUR
250 x 4 mm
C18 Pyramid, 5 m
A) 50 mM NH4H2PO4, pH 2.5; B) acetonitrile
0 60% B in 13 min
1.0 ml/min
25 C
UV, 230 nm (4.5 min)
UV, 261 nm
4 l

(50:50, v/v)
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj. volume:
1.0 l
Column:
Eluent:
MN Appl. No. 119110
Appl. 14: Cold medicine (purchasable OTC drug)

Peaks:
1. Ascorbic acid
2. Paracetamol
3. Caffeine
4. Chlorpheniramine
MN Appl. No. 119320
Appl. 16: Antibacterial drugs

Peaks:
1. Ofloxacin
2. Ciprofloxacin
3. Cinoxacin
4. Penicillin G
5. Penicillin V
6. Cloxacillin
8.0
3
6
1
9.5
5
4
2
12 min

A) 50 mM NH4H2PO4, pH 2.5; B) acetonitrile
0 60% B in 13 min
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 230 nm (4.5 min)
UV, 261 nm
Inj. volume:
2 l
Column:
Eluent:
MN Appl. No. 119120
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
12
min

acetonitrile water (40:60, v/v) 0.05% TFA
1.0 ml/min
25 C
UV, 254 nm
5 l
MN Appl. No. 117870
19
Applications
Appl. 17: Quinolone antibiotics
Appl. 19: Penicillin antibiotics
Peaks:
1. Cinoxacin
2. Oxolinic acid
3. Nalidixic acid
Peaks:
1. Amoxicillin
2. Penicillin G
3. Penicillin V
4. Nafcillin
5. Dicloxacillin
1
4
3
3
2
min
10
150 x 4.6 mm NUCLEODUR Sphinx RP, 5 m

methanol 20 mM KH2PO4, pH 2,5
(50:50, v/v)
Flow rate:
1.0 ml/min
Temperature: ambient
Detection:
UV, 254 nm
Inj. volume:
5 l
Column:
Eluent:
2.5
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
7.5
min

acetonitrile 0.1% TFA (50:50, v/v)
1.0 ml/min
25 C
UV, 254 nm
1 l
MN Appl. No. 119870
MN Appl. No. 119150
Appl. 18: Penicillin antibiotics
Appl. 20: Sulfonamides
Peaks:
1. Penicillin G
2. Penicillin V
3. Cloxacillin
4. Nafcillin
5. Dicloxacillin
Peaks:
1. Sulfanilamide
2. Sulfadiazine
3. Sulfathiazole
4. Sulfamerazine
5. Sulfadimidine
6. Succinylsulfathiazole
1
2
1
4
3
4
5
2
6 min

(40:60, v/v)
Flow rate:
1.0 ml/min
Temperature: 22 C
Detection:
UV, 254 nm
Inj. volume:
5 l
Column:
Eluent:
MN Appl. No. 117860
20
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
min
125 x 4 mm NUCLEODUR Sphinx RP, 5 m

methanol 0.1% TFA (20:80, v/v)
1.0 ml/min
22 C
UV, 230 nm
3 l
MN Appl. No. 119860
Applications
Appl. 21: Sulfonamides
Appl. 23: Sedative drugs
Peaks:
1. Sulfanilamide
2. Sulfadiazine
3. Sulfathiazole
4. Sulfamerazine
5. Sulfadimidine
6. Succinylsulfathiazole
23
4
Peaks:
1. Promethazine
2. Promazine
3. Chlorpromazine
1
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
0
0
10 min
10
Column:
Eluent:

methanol 0.1% TFA (20:80, v/v)
1.0 ml/min
22 C
UV, 230 nm
4 l
15
125 x 4 mm
100-5 CN-RP
A) methanol
B) 50 mM ammonium acetate, pH 5.0
70 50% B in 10 min, then 10 min 50% B
Flow rate:
1.5 ml/min
Temperature: 30 C
Detection:
UV, 254 nm
Inj. volume:
1 l (670 g/ml 1 + 2, 335 g/ml 3)
MN Appl. No. 119300
MN Appl. No. 117880
Appl. 22: Benzodiazepines

Peaks:
1. Bromazepam
2. Oxazepam
3. Lorazepam
4. Temazepam
Appl. 24: Coronary therapeutic drugs

(Ca-antagonists)
3
1
min
NUCLEODUR
Peaks:
1. Nifedipine
2. Nitrendipine
3. Nimodipine
4. Nisoldipine
5. Nicardipine
4
2
3
2
4
5
min

(45:55, v/v)
Flow rate:
1.0 ml/min
Temperature: 22 C
Detection:
UV, 254 nm
Inj. volume:
5 l
Column:
Eluent:
MN Appl. No. 117850
Column:
Eluent:
12 min
NUCLEODUR
125 x 4 mm
100-5 CN-RP
A) acetonitrile, B) 20 mM KH2PO4, pH 6.5
30 50% B in 7.5 min, then 7.5 min 50% B
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj. volume:
2.5 l (25 g/compound)
MN Appl. No. 119310
21
Applications
Appl. 25: Nucleic acid bases
Appl. 27: Alkaloids
Peaks:
1. Cytosine
2. Adenine
3. Uracil
4. Thymine
3
2
1
Peaks:
1. Codeine
2. Quinine
3. Strychnine
4. Atropine
5. Papaverine
6. Noscapine
3
6
5
2
min
12
10
15
min

A) 50 mM NH4H2PO4, pH 2.5, B) acetonitrile
2.5 min 100% A, then to 90% A in 10 min
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
Inj. volume:
3 l

A) acetonitrile, B) 20 mM KH2PO4, pH 2.5
90 70% B in 25 min, then 70 90% B in 1 min
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 210 nm
Inj. volume:
5 l
MN Appl. No. 119140
MN Appl. No. 117950
Column:
Eluent:
Column:
Eluent:
Appl. 28: Catecholamines
Appl. 26: Quinine alkaloids

Peaks:
1. Chloroquine
2. Quinine
3. Mefloquine
1
1
Peaks:
1. Norephedrine
2. Adrenaline
3. Dihydroxyphenylalanine
4. Hydroxytyramine
5. Tyrosine
Column:
Eluent:
min
10
NUCLEODUR
125 x 4 mm
C18 Gravity, 5 m
A) methanol, B) 20 mM KH2PO4, pH 2.5
90 70% B in 4 min, then 70 30% B in 7 min
Flow rate:
1.3 ml/min
Detection:
UV, 240 nm
Temperature: 25 C
Inj. volume:
10 l
MN Appl. No. 117960
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
min

100 mM NaH2PO4, pH 3.0
0.8 ml/min
25 C
UV, 254 nm
5 l
MN Appl. No. 117930
22
Applications
Appl. 29: Steroids
Peaks:
1. Cortisone
2. Hydrocortisone
3. Hydrocortisone
21-acetate
4. 6-Methyl-11hydroxyprogesterone
acetate
Appl. 31: Steroids

6
1 2
Peaks
(each env. 10 50 g/ml):
1.Estriol
2.Prednisolone
3.Cortisone
4.Testosterone
5.6-Methyl-11-hydroxyprogesterone
6.6-Methyl-17-hydroxyprogesterone
7.6-Methyl-17-hydroxyprogesterone acetate
8.Estradiol
9.Estrone
10.Progesterone
4
5
Column:
Eluent:
Flow rate:
Temperature:
Detection:
10
min

1.0 ml/min
25 C
UV, 240 nm
10
3
5 7
10
15
20 min 25
NUCLEODUR
Column:
Eluent:
125 x 4 mm
100-5 C8 ec
A) water, B) methanol
1 min 20% B, 20 35% B in 10 min, 3 min 35%
B, 35 60% B in 6 min, 5 min 60% B
Flow rate:
1.0 ml/min
Temperature: 30 C
Detection:
UV, 230 nm
Inj. volume:
10 l
MN Appl. No. 118550
Appl. 30: Corticosteroids
Appl. 32: Xylometazoline in nasal spray

1
Peaks:
1. Xylometazoline
2. 5. Benzalkonium chlorides
with different chain lengths
(C8, C10, C12, C14)
a) standard
b) nasal spray
9
6
MN Appl. No. 118540
Peaks:
1. Cortisone
2. Hydrocortisone
3. Corticosterone
4. Hydrocortisone
acetate
5. Corticosterone
acetate
4
3
2
b)
a)
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
15
min

1.0 ml/min
25 C
UV, 230 nm
5 l
MN Appl. No. 117920
0
Column:
Eluent:
Flow rate:
Detection:
min

acetonitrile 50 mM Na citrate, pH 3.0 (50:50, v/v)
0.8 ml/min; temperature: 40 C
UV, 254 nm
MN Appl. No. 120390
23
Applications
Appl. 33: Soft drink additives
Peaks:
1. Ascorbic acid
2. Acesulfam K
3. Saccharin
4. Caffeine
5. Aspartame
6. Quinine
7. Vanillin
8. Sorbic acid
9. Benzoic acid
Appl. 35: Aromatic aldehydes

Peaks:
1. p-Carboxybenzaldehyde
2. p-Hydroxybenzaldehyde
3. Vanillin
4. 4-Ethoxyvanillin
5. Benzaldehyde
(acidic)
(basic)
(strongly basic)
3
2
(acidic)
(acidic)
1
3
67
10
min
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
NUCLEODUR
150 x 4.6 mm
100-5 C8 ec
20 mM KH2PO4, pH 3 acetonitrile (5:1, v/v)
1.9 ml/min
25 C
UV, 220 nm
10 l
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
125 x 4 mm
100-5 C18 ec
acetonitrile water, pH 6.0 (22:78, v/v)
1.0 ml/min
22 C
UV, 254 nm
5 l
MN Appl. No. 118560
MN Appl. No. 117990
Appl. 34: Fast separation of sweeteners
Appl. 36: Substituted aromatics
Peaks:
1. Acesulfam K
2. Saccharin
3. Aspartame
min
NUCLEODUR
Peaks:
1. Uracil
2. Benzamide
3. Phenol
4. Benzaldehyde
5. Acetophenone
6. 2-Nitrophenol
7. Nitrobenzene
45
3 7
8
10
9
6
1
11
8. Propyl 4-hydroxybenzoate
9. Toluene
10. Benzophenone
11. Xylene
10
11
A) C18 Gravity
B) Sphinx RP
Column:
50 x 4 mm
NUCLEODUR
100-5 C18 ec
Eluent:
acetonitrile water
(20:80, v/v)
0.1% TFA
Flow rate:
2.5 ml/min
Temperature: 25 C
Detection:
UV, 215 nm
Inj. volume:
10 l
MN Appl. No. 117940
24
min
Columns:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
10
15
20
25
min
150 x 4.6 mm NUCLEODUR

1.0 ml/min
40 C
UV, 254 nm
2 l
MN Appl. No. 119840/119850
Applications
Appl. 37: Separation of a test mixture
Appl. 39: Fat-soluble vitamins
Peaks:
1. Ethyl p-aminobenzoate
2. Propranolol
3. Lidocaine
4. Imipramine
5. Amitriptyline
6. Trimipramine
1
5
Peaks:
1. Vitamin K3
2. Vitamin A
3. Vitamin A acetate
4. Vitamin D2
5. Vitamin D3
6. Vitamin E
7. Vitamin E acetate
8. Vitamin K1
3
6
23
0
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
10
10
6 7
15
min
min

1.0 ml/min
30 C
UV, 254 nm
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:

acetonitrile
1.5 ml/min
30 C
UV, 280 nm
4 l
MN Appl. No. 118620
MN Appl. No. 117890
Appl. 38: Selectivity test

Peaks (inj. volume 6 l):
1. Uracil
4. Ethyl benzoate
5. Lidocaine
6. Biphenyl
7. Acenaphthene
Appl. 40: Tocopherols

Peaks:
1. -Tocopherol
2. -Tocopherol
3. -Tocopherol
4. -Tocopherol acetate
1
3
2
4
3
7
1
0
0
Column:
Eluent:
Flow rate:
Temperature:
Detection:
min
6 min
10
125 x 4 mm NUCLEODUR Sphinx RP, 5 m

methanol 25 mM NH4H2PO4, pH 7 (65:35, v/v)
1.0 ml/min
40 C
UV, 254 nm
MN Appl. No. 119880
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:

methanol (100%)
1.0 ml/min
22 C
UV, 295 nm
8 l
MN Appl. No. 117910
25
Applications
2
Appl. 41: Water-soluble vitamins
Appl. 42: Complexing agents

acc. to DIN 38 413-8
Peaks: (mg/ml)
1. Nicotinic acid (0.12)
2. Nicotinamide (0.12)
3. 4-Aminobenzoic acid (0.03)
4. Folic acid (0.24)
5. Pyridoxin (B6) [hydrochloride] (0.06)
6. Riboflavin (B2) (0.012)
7. Thiamine hydrochloride (B1) (0.06)
8. Rutin (0.012)
red curves: vitamin test mixture (in eluent A)
blue curves: multivitamin juice (undiluted)
Peaks:
1. NTA (nitrilotriacetic acid)
2. EDTA (ethylenediamintetraacetic acid)
3. DTPA (diethylenetrinitrilopentaacetic
acid)
chromatogram courtesy of H. Albrich,
C. Geis, Gewerbliches Institut fr Umweltanalytik GmbH, Teningen, Germany
7 8
254 nm
10
14
18
22
min

0.6 mmol/l HNO3, 7.53 mmol/l N(C4H9)4HSO4,
2.6 mmol/l N(C4H9)4OH, 37 mol/l Fe3+
Flow rate:
0.6 ml/min
Temperature: 20 C
Detection:
UV, 260 nm
Inj. volume:
50 l
MN Appl. No. 119780
Column:
Eluent:
275 nm
Appl. 43: Phenolic compounds

Peaks:
1. Resorcinol
2. Pyrocatechol
3. 4-Methoxyphenol
4. Phenol
5. 2-Methoxyphenol
6. 2-Ethoxyphenol
1
4 5
361 nm
Column:
Eluent:
10
15
20
min
25
NUCLEODUR
125 x 4 mm
C18 Pyramid, 5 m
A) water, 15 mM heptanesulfonic acid (Na salt),
25 mM NaH2PO4, 0.25% CH3COOH,
0.005% triethylamine (pH 3.5),
B: acetonitrile water (40:60, v/v), 15 mM
heptanesulfonic (Na salt), 0.25 % CH3COOH,
0.005% triethylamine (pH ~ 3.5); multistep
gradient: 5 min 0% B, 0 10% B in 2.5 min,
10 25% B in 2.5 min, 25 50% B in 8 min,
50 70% B in 7 min, 70 0%B in 1 min
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254, 275 and 361 nm
Inj. volume:
10 l
MN Appl. No. 119770
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
min
10

methanol water, 0.1% H3PO4 (40:60, v/v)
1.0 ml/min
22 C
UV, 254 nm
5 l
MN Appl. No. 117970
26
Applications
Appl. 44: Organic acids
Peaks:
1. Aspartic acid
2. Fumaric acid
3. Maleic acid
Appl. 46: Pesticides

Peaks:
1. Desisopropylatrazine
2. Metamitron
3. Desethylatrazine
4. Bromoxynil
5. Simazine
6. Cyanazine
7. Metabenzthiazuron
8. Atrazine
9. Monolinuron
1
2
10.Isoproturon
11.Diuron
12.Metobromuron
13.Metazachlor
14.Sebutylazine
15.Terbuthylazine
16.Linuron
17.Chloroxuron
18.Metolachlor
1
2
Column:
Eluent:
Flow rate:
Temperature:
Detection:
Inj. volume:
2.5
7.5
min
7 8

25 mM KH2PO4, pH 4.0
0.5 ml/min
30 C
UV, 210 nm
15 l
15
17 18
5
12
MN Appl. No. 119290
Appl. 45: Test for metal ions in silica adsorbent

1
Peaks:
14
10
20
10
9 11 13
30
16
40
50
60 min

A) water (1% acetic acid), B) acetonitrile
10 25% B in 10 min, 25 30% B in 10 min,
5 min at 30% B, 30 40 % B in 20 min
40 50% B in 10 min, 10 min at 50% B
Flow rate
1.0 ml/min
Temperature: 35 C
Detection
UV 230 nm
Column:
Eluent:
Gradient:
MN Appl. No. 118610
min
The ratio of the asymmetry factors of 2,3-dihydroxynaphthalene

(2) and 2,7-dihydroxynaphthalene (1) is a measure for the metal
ion content of the silica phase, because (2) can form complexes
with metal ions, resulting in broad peaks for this compound.
Column:
Eluent:
Flow rate:
1.0 ml/min
Temperature: 25 C
Detection:
UV, 254 nm
MN Appl. No. 118630
27
Alphabetical Index of Analytes

Analyte
NUCLEODUR phase
page
A
Acenaphthene
Acetanilide
Acetic acid
Acetophenone
Acetylsalicylic acid
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 C18 ec
100-5 C8 ec
C8 Gravity, 5 m
100-5 CN-RP
C18 Gravity, 5 m
C8 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 C18 ec
Adenine
Adrenaline
4-Aminobenzoic acid
Amitriptyline
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
Acesulfam K
2-Acetamidophen
Acetaminophen
C18 Pyramid, 5 m
C8 Gravity, 5 m
Amoxicillin
Aniline
Anthracene
Ascorbic acid
Aspartame
Aspartic acid
Atrazine
Atropine
C18 Pyramid, 5 m
C18 Pyramid, 5 m
100-5 C18 ec
100-5 C8 ec
C18 Pyramid, 5 m
100-5 C18 ec
100-5 C8 ec
100-5 CN-RP
100-3 C8 ec
C18 Gravity, 5 m
Fig. 5, 7
Fig. 19
Appl. 38
Appl. 34
Appl. 33
Appl. 4
Fig. 25
Appl. 12
Appl. 4
Fig. 14, 16
Appl. 36
Appl. 1, 2
Appl. 6
Appl. 5
Appl. 7
Appl. 3
Appl. 4
Appl. 25
Appl. 28
Appl. 41
Fig. 5, 7
Appl. 9, 10
Fig. 18
Appl. 11
Appl. 37
Appl. 19
Fig. 17
Fig. 3
Appl. 33
Appl. 13, 14
Appl. 34
Appl. 33
Appl. 44
Appl. 46
Appl. 27
4
10
25
24
24
16
14
18
16
9
24
16
17
17
17
16
16
22
22
26
4
18
10
18
25
20
10
3
24
19
24
24
27
27
22
B
100-5 C18 ec
Sphinx RP, 5 m
Benzalkonium chlorides 100-5 CN-RP
Benzamide
100-5 CN-RP
Benzophenone
Biphenyl
Biphenyl-2-ol
Bromazepam
Bromoxynil
Brompheniramine
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 C8 ec
100-5 C18 ec
100-3 C8 ec
100-5 CN-RP
Appl. 35
Appl. 36
Appl. 32
Fig. 24
Fig. 26
Fig. 13
Fig. 19
Appl. 36
Appl. 33
Appl. 12
Fig. 19
Appl. 36
Fig. 24
Fig. 26
Fig. 19
Appl. 38
Fig. 21
Appl. 22
Appl. 46
Fig. 25
24
24
23
14
15
7
10
24
24
18
10
24
14
15
10
25
11
21
27
14
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
Sphinx RP, 5 m
Appl. 33
Fig. 11
Appl. 13, 14
Appl. 4
Fig. 22
24
6
19
16
12
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 CN-RP
28
p-Carboxybenzaldehyde
Carprofen
Catechin
Chloroquine
Chloroxuron
Chlorpheniramine
100-5 C18 ec
100-5 C8 ec
Sphinx RP, 5 m
C18 Gravity, 5 m
100-3 C8 ec
100-5 CN-RP
C18 Gravity, 5 m
C18 Pyramid, 5 m
Chlorpromazine
Cinoxacin
100-5 CN-RP
100-5 C18 ec
Sphinx RP, 5 m
100-5 C18 ec
100-5 CN-RP
C18 Gravity, 5 m
100-5 CN-RP
100-5 C18 ec
Ciprofloxacin
Clenbuterol
Clomipramine
Clonidin
Cloxacillin
Codeine
Corticosterone acetate
Corticosterone
Cortisone
Cyanazine
Cytosine
page
Appl. 35
Fig. 20
Fig. 23
Appl. 26
Appl. 46
Fig. 25
Appl. 12
Fig. 19
Appl. 13, 14
Appl. 23
Appl. 16
Appl. 17
Appl. 16
Appl. 15
Appl. 9
Appl. 15
Appl. 16
Appl. 18
Appl. 27
Appl. 30
Appl. 30
Appl. 30
Appl. 31
Fig. 27
Appl. 29
Appl. 46
Appl. 25
24
11
13
22
27
14
18
10
19
21
19
20
19
19
18
19
19
20
22
23
23
23
23
15
23
27
22
Appl. 46
Appl. 46
Appl. 12
Appl. 13
Fig. 5, 7
Appl. 1
Appl. 6
Fig. 20
Appl. 18
Appl. 19
27
27
18
19
4
16
17
11
20
20
Appl. 42
Appl. 5
Appl. 7
26
17
17
C18 Pyramid, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
C18 Pyramid, 5 m
100-3 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
C18 Pyramid, 5 m
Fig. 7
Appl. 45
Appl. 38
Appl. 28
Fig. 24
Fig. 26
Fig. 19
Fig. 17
Fig. 12
Appl. 13
Appl. 46
Appl. 9, 10
Fig. 18
Appl. 11
Appl. 42
4
27
25
22
14
15
10
10
7
19
27
18
10
18
26
C18 Pyramid, 5 m
100-5 CN-RP
100-5 C8 ec
100-5 C8 ec
Appl. 42
Fig. 25
Appl. 31
Appl. 31
26
14
23
23
C18 Gravity, 5 m
100-5 C18 ec
100-5 C18 ec
100-5 C18 ec
100-5 C8 ec
100-5 CN
C8 Gravity, 5 m
100-3 C8 ec
C18 Pyramid, 5 m
D
Desethylatrazine
Desisopropylatrazine
Dextromethorphan
Dibutyl phthalate
Diclofenac
100-3 C8 ec
100-3 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
100-5 C18 ec
100-5 C8 ec
100-5 C18 ec
C18 Pyramid, 5 m
Diethylenetrinitrilopentaacetic acid
Diflunisal
C18 Pyramid, 5 m
100-5 C8 ec
C18 Gravity, 5 m
Dihydroxynaphthalene isomers
C18 Gravity, 5 m
C8 Gravity, 5 m
Sphinx RP, 5 m
Dihydroxyphenylalanine C18 Gravity, 5 m
Dimethyl phthalate
100-5 CN-RP
Dimethylaniline
Diphenhydramine
Diuron
Doxepin
DTPA
C
Caffeine
NUCLEODUR phase
Dicloxacillin
Benzaldehyde
Benzoic acid
Analyte
E
EDTA
Ephedrine
Estradiol
Estriol

Analyte
NUCLEODUR phase
Estrone
2-Ethoxyphenol
100-5 C8 ec
100-5 C18 ec
100-5 C8 ec
100-5 C18 ec
C18 Pyramid, 5 m
C8 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
page
C18 Pyramid, 5 m
Sphinx RP, 5 m
Appl. 31
Appl. 43
Fig. 21
Appl. 35
Fig. 19
Appl. 37
Fig. 17
Fig. 7
Fig. 10
Fig. 17
Appl. 38
23
26
11
24
10
25
10
4
5
10
25
C18 Pyramid, 5 m
Appl. 42
26
Fenoprofen
100-5 C8 ec
Fisetin
Flurbiprofen
Sphinx RP, 5 m
100-5 C18 ec
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Pyramid, 5 m
C18 Pyramid, 5 m
100-5 CN-RP
Fig. 20
Appl. 5
Fig. 23
Appl. 6
Appl. 5
Appl. 7
Appl. 3
Appl. 41
Fig. 14
Appl. 44
11
17
13
17
17
17
16
26
9
27
Appl. 8
Appl. 30
Fig. 27
Appl. 29
Appl. 30
Appl. 29
Appl. 35
17
23
15
23
23
23
24
Appl. 3
Appl. 28
16
22
Appl. 1
Appl. 6
Appl. 5
Appl. 7
Appl. 3
Appl. 9, 10
Fig. 18
Appl. 11
Appl. 37
Appl. 5
Appl. 7
Appl. 46
Fig. 23
16
17
17
17
16
18
10
18
25
17
17
27
13
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
Fig. 23
Appl. 1
Appl. 6
Fig. 20
Fig. 13
Appl. 3
13
16
17
11
7
16
C18 Pyramid, 5 m
Fig. 15
C18 Gravity, 5 m
Fig. 7 4, Fig. 127,
4-Ethoxyvanillin
Ethyl benzoate
Ethyl p-aminobenzoate
p-Ethylaniline
Ethylbenzene
Ethyl benzoate
Ethylenediaminetetraacetic acid
Folic acid
Formic acid
Fumaric acid
H
Hexobarbital
Hydrocortisone
100-5 C18 ec
100-5 C18 ec
100-5 CN
C8 Gravity, 5 m
Hydrocortisone acetate 100-5 C18 ec
C8 Gravity, 5 m
4-Hydroxybenzaldehyde 100-5 C18 ec
4-Hydroxymethyl
benzoate
C18 Pyramid, 5 m
Hydroxytyramine
C18 Gravity, 5 m
I
Ibuprofen
Imipramine
Indomethazin
Isoproturon
Isorhamnetin
100-5 C18 ec
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
100-5 C8 ec
C18 Gravity, 5 m
100-3 C8 ec
Sphinx RP, 5 m
K
Kaempferol
Ketoprofen
Sphinx RP, 5 m
100-5 C18 ec
L
Lactic acid
Lidocaine
Analyte
Fig. 13
Linuron
Lorazepam
NUCLEODUR phase
7
C18 Pyramid, 5 m
C8 Gravity, 5 m
Sphinx RP, 5 m
100-3 C8 ec
100-5 C18 ec
page
Fig. 19
Appl. 37
Appl. 38
Appl. 46
Appl. 22
10
25
25
27
21
M
Maleic acid
44
Malic acid
Maprotiline
Meclofenamic acid
Medrysone
Mefloquine
Mephobarbital
Metabenzthiazuron
Metamitron
Metazachlor
Methoxyphenol isomers
100-5 CN-RP
27
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
100-5 C18 ec
100-5 CN
C18 Gravity, 5 m
100-5 C18 ec
100-3 C8 ec
100-3 C8 ec
100-3 C8 ec
100-5 C18 ec
100-5 C8 ec
6-Methyl-11-hydroxyprogesterone
100-5 C8 ec
C8 Gravity, 5 m
6-Methyl-17-hydroxyprogesterone
100-5 C8 ec
C8 Gravity, 5 m
6-Methyl-17-hydroxyprogesterone acetate
100-5 C8 ec
C8 Gravity, 5 m
Methyltestosterone
100-5 CN
Metobromuron
100-3 C8 ec
Metolachlor
100-3 C8 ec
Monolinuron
100-3 C8 ec
Fig. 25 14, Appl.

Appl. 12
Fig. 16
Fig. 15
Appl. 9
Appl. 6
Fig. 27
Appl. 26
Appl. 8
Appl. 46
Appl. 46
Appl. 46
18
9
9
18
17
15
22
17
27
27
27
Appl. 43
Fig. 21
26
11
Appl. 31
Appl. 29
23
23
Appl. 31
Appl. 29
23
23
Appl. 31
Appl. 29
Fig. 27
Appl. 46
Appl. 46
Appl. 46
23
23
15
27
27
27
Appl. 18
Appl. 19
Appl. 17
Fig. 3
Fig. 7
Fig. 19
Appl. 5
Appl. 7
Appl. 24
Appl. 41
Appl. 41
Appl. 24
Appl. 24
Appl. 24
Appl. 24
Appl. 42
Appl. 36
Appl. 36
Appl. 9
Appl. 9
Fig. 25
Appl. 28
Fig. 27
20
20
20
3
4
10
17
17
21
26
26
21
21
21
21
26
24
24
18
18
14
22
15
N
Nafcillin
Nalidixic acid
Naphthalene
Naproxen
Nicardipine
Nicotinamide
Nicotinic acid
Nifedipine
Nimodipine
Nisoldipine
Nitrendipine
Nitrilotriacetic acid
Nitrobenzene
2-Nitrophenol
Norclomipramine
Nordoxepin
Norephedrine
Norgestrel
100-5 C18 ec
C18 Pyramid, 5 m
Sphinx RP, 5 m
100-5 C18 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
100-5 C8 ec
C18 Gravity, 5 m
100-5 CN-RP
C18 Pyramid, 5 m
C18 Pyramid, 5 m
100-5 CN-RP
100-5 CN-RP
100-5 CN-RP
100-5 CN-RP
C18 Pyramid, 5 m
Sphinx RP, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
C18 Gravity, 5 m
100-5 CN-RP
C18 Gravity, 5 m
100-5 CN
29
Alphabetical index of analytes

Analyte
NUCLEODUR phase
Nortriptyline
Noscapine
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
C18 Gravity, 5 m
NTA
page
C18 Pyramid, 5 m
Appl. 9, 10
Fig. 18
Appl. 11
Fig. 12
Appl. 27
Appl. 42
18
10
18
7
22
26
100-5 C18 ec
100-5 C18 ec
Sphinx RP, 5 m
Appl. 16
Appl. 22
Appl. 17
19
21
20
Papaverine
C18 Gravity, 5 m
Paracetamol
100-5 C18 ec
C18 Pyramid, 5 m
Fig. 12
Appl. 27
Appl. 1
Appl. 3
Appl. 13, 14
Appl. 4
Appl. 16
Appl. 18
Appl. 19
Appl. 16
Appl. 18
Appl. 19
Appl. 8
Appl. 4
Fig. 21
Fig. 24
Fig. 26
Appl. 8
Fig. 3
Appl. 43
Fig. 21
Fig. 6
Fig. 17
Fig. 22
Appl. 36
Fig. 20
Appl. 5
Appl. 7
Appl. 31
Fig. 27
Appl. 15
Appl. 31
Appl. 23
Appl. 23
Appl. 37
7
22
16
16
19
16
19
20
20
19
20
20
17
16
11
14
15
17
3
26
11
4
10
13
24
11
17
17
23
15
19
23
21
21
25
Sphinx RP, 5 m
C18 Pyramid, 5 m
100-5 C18 ec
100-5 C8 ec
Appl. 36
Appl. 9, 10
Fig. 18
Appl. 11
Appl. 12
Fig. 6
Fig. 10
Fig. 22
Appl. 41
Appl. 43
Fig. 21
24
18
10
18
18
4
5
13
26
26
11
Sphinx RP, 5 m
100-5 C8 ec
C18 Gravity, 5 m
Fig. 23
Appl. 33
Appl. 26, 27
13
24
22
O
Ofloxacin
Oxazepam
Oxolinic acid
Penicillin G
C8 Gravity, 5 m
100-5 C18 ec
Penicillin V
C18 Pyramid, 5 m
100-5 C18 ec
Pentobarbital
Phenactin
Phenetole
C18 Pyramid, 5 m
100-5 C18 ec
C8 Gravity, 5 m
100-5 C8 ec
100-5 CN-RP
Phenobarbital
Phenol
100-5 C18 ec
100-5 C18 ec
Phenol
Piroxicam
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
Sphinx RP, 5 m
100-5 C8 ec
Procainamide
Progesterone
Promazine
Promethazine
Propranolol
Propyl 4-hydroxy
benzoate
Protriptyline
Pseudoephedrine
Pyridine
Pyridoxin
Pyrocatechol
C18 Gravity, 5 m
100-5 C8 ec
100-5 CN
100-5 CN-RP
100-5 C8 ec
100-5 CN-RP
100-5 CN-RP
C8 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
C18 Pyramid, 5 m
C8 Gravity, 5 m
C18 Gravity, 5 m
C18 Gravity, 5 m
Q
Quercetin
Quinine
30
NUCLEODUR phase
page
R
Resorcinol
Riboflavin
Rutin
100-5 C18 ec
100-5 C8 ec
C18 Pyramid, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
Appl. 43
Fig. 21
Appl. 41
Appl. 41
Fig. 23
26
11
26
26
13
100-5 C18 ec
100-5 C8 ec
100-5 C18 ec
100-3 C8 ec
100-3 C8 ec
100-5 C8 ec
C18 Gravity, 5 m
C18 Pyramid, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
Sphinx RP, 5 m
100-5 C18 ec
C18 Gravity, 5 m
100-5 C8 ec
Appl. 34
Appl. 33
Appl. 1, 2
Appl. 46
Appl. 46
Appl. 33
Appl. 27
Fig. 15
Appl. 21
Appl. 20
Appl. 21
Appl. 20
Appl. 21
Appl. 20
Appl. 21
Appl. 20
Appl. 21
Appl. 20
Appl. 21
Appl. 20
Appl. 6
Appl. 7
Fig. 20
Appl. 5
Appl. 7
24
24
16
27
27
24
22
9
21
20
21
20
21
20
21
20
21
20
21
20
17
17
11
17
17
Fig. 15, 16
Appl. 22
Appl. 46
Appl. 31
Fig. 27
Fig. 11
Fig. 22
Appl. 41
Appl. 8
Appl. 25
Appl. 40
Appl. 6
Appl. 5
Appl. 7
Fig. 7
Fig. 10
Fig. 17
Appl. 36
Appl. 9, 10
Appl. 11
Appl. 37
Appl. 28
9
21
27
23
15
6
12
26
17
22
25
17
17
17
4
5
10
24
18
18
25
22
Fig. 7
Fig. 17
Appl. 25
Appl. 36
Appl. 38
4
10
22
24
25
Prednisolone
Analyte
Saccharin
Salicylic acid
Sebutylazine
Simazine
Sorbic acid
Strychnine
Succinic acid
Succinylsulfathiazole
Sulfadiazine
Sulfadimidine
Sulfamerazine
Sulfanilamide
Sulfathiazole
Sulindac
Suprofen
C18 Gravity, 5 m
T
Tartaric acid
Temazepam
Terbuthylazine
Testosterone
Theophylline
Thiamine
Thiamylal
Thymine
Tocopherols
Tolmetin
Toluene
Trimipramine
Tyrosine
C18 Pyramid, 5 m
100-5 C18 ec
100-3 C8 ec
100-5 C8 ec
100-5 CN
C18 Gravity, 5 m
Sphinx RP, 5 m
C18 Pyramid, 5 m
100-5 C18 ec
C18 Pyramid, 5 m
100-5 C18 ec
100-5 C18 ec
100-5 C8 ec
C18 Gravity, 5 m
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m
C18 Gravity, 5 m
C8 Gravity, 5 m
C18 Gravity, 5 m
U
Uracil
C18 Gravity, 5 m
C18 Pyramid, 5 m
Sphinx RP, 5 m

NUCLEODUR phase
Analyte
page
NUCLEODUR phase
Analyte
page
Vanillin
Veratrol
Vitamins
fat-soluble
water-soluble
100-5 C18 ec
100-5 C8 ec
100-5 C8 ec
Appl. 35
Appl. 33
Fig. 21
24
24
11
100-5 C18 ec
C18 Pyramid, 5 m
Appl. 39
Appl. 41
25
26
o-Xylene
100-5 CN-RP
Xylene
Xylometazoline
Sphinx RP, 5 m
100-5 CN-RP
Fig. 24
Fig. 26
Appl. 36
Appl. 32
14
15
24
23
Ordering Information
Analytical columns with NUCLEODUR C18 Gravity
30 mm 1)
Length
50 mm
70 mm
100 mm
125 mm
150 mm
250 mm
Guard
columns

Particle size 3 m, pore size 110 ; octadecyl phase, endcapped, 18% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID 761450.20
3 mm ID 761450.30
4 mm ID 761450.40
4.6 mm ID 761450.46
Microbore columns
1 mm ID
761451.20
761451.30
761451.40
761451.46
761452.20
761452.30
761452.40
761452.46
761454.46
761453.20
761453.30
761453.40
761453.46
717715.10
717716.10
717717.10
760083.46
760082.20
760082.30
760082.40
760082.46
761124.30
761124.30
761124.40
761124.40
2)
EC columns 3)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
NUCLEODUR C18
717714.10
760080.20
760080.30
760080.40
760080.46
760081.20
760081.30
760081.40
760081.46
761124.30
761124.30
761124.40
761124.40
Gravity, 5 m
Particle size 5 m, pore size 110 ; octadecyl phase, endcapped, 18 % C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID 761502.20
3 mm ID 761502.30
4 mm ID 761502.40
4.6 mm ID 761502.46
761503.20
761503.30
761503.40
761503.46
Microbore columns 2)
1 mm ID
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
717706.10
761500.20
761500.30
761500.40
761500.46
761504.46
761501.20
761501.30
761501.40
761501.46
717707.10
717708.10
717705.10
760103.46
760101.20
760101.30
760101.40
760101.46
761125.30
761125.30
761125.40
761125.40
3)
760102.20
760102.30
760102.40
760102.46
760100.20
760100.30
760100.40
760100.46
761125.30
761125.30
761125.40
761125.40
30 mm ChromCart columns and CC guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.
1) 30 mm ChromCart columns require CC column holder 30 mm (Cat. No. 721823).
2) On request, Microbore columns are also available in lengths of 40, 60, 200 and 300 mm and with 0.15, 0.3, 0.4, 0.5, 0.75 and 1.5 mm ID.
Guard columns for Microbore columns on request.
3) As guard columns for EC columns use ChromCart guard column cartridges with guard column adaptor EC (Cat. No. 721359).
Preparative columns with NUCLEODUR phases are available on request!

31
Analytical columns with NUCLEODUR C18 Pyramid
Length
50 mm
70 mm
100 mm
125 mm
150 mm
250 mm
NUCLEODUR C18 Pyramid, 3 m
Guard
columns
NEW!
Particle size 3 m, pore size 110 ; octadecyl phase with hydrophilic endcapping, 14% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
Microbore columns
1 mm ID
761853.20
761853.30
761853.40
761853.46
761850.20
761850.30
761850.40
761850.46
761851.46
761852.20
761852.30
761852.40
761852.46
717742.10
717743.10
717744.10
760260.20
760260.30
760260.40
760260.46
760261.20
760261.30
760261.40
760261.46
760262.20
760262.30
760262.40
760262.46
761854.30
761854.30
761854.40
761854.40
1)
717740.10
EC columns 2)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
717741.10
760263.20
760263.30
760263.40
760263.46
761854.30
761854.30
761854.40
761854.40
NUCLEODUR C18 Pyramid, 5 m

Particle size 5 m, pore size 110 ; octadecyl phase with hydrophilic endcapping, 14% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
761801.20
761801.30
761801.40
761801.46
1 mm ID
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
717722.10
761802.20
761802.30
761802.40
761802.46
761803.46
761803.20
761803.30
761803.40
761803.46
717723.10
717724.10
717725.10
760203.46
760202.20
760202.30
760202.40
760202.46
761800.30
761800.30
761800.40
761800.40
2)
760200.20
760200.30
760200.40
760200.46
760201.20
760201.30
760201.40
760201.46
Analytical columns with NUCLEODUR Sphinx RP

NUCLEODUR
761800.30
761800.30
761800.40
761800.40
NEW!
Sphinx RP, 3 m
Particle size 3 m, pore size 110 ; special bifunctional RP phase, 14% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
EC columns 2)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
760806.20
760806.30
760806.40
760806.46
761556.20
761556.30
761856.40
761856.46
761558.46
760807.20
760807.30
760807.40
760807.46
760805.20
760805.30
760805.40
760805.46
761557.30
761557.30
761557.40
761557.40
760808.20
760808.30
760808.40
760808.46
761557.30
761557.30
761557.40
761557.40
ChromCart guard column cartridges (8 mm) in packs of 3, all other columns in packs of 1.
32
Analytical columns with NUCLEODUR Sphinx RP (cont.)
30 mm 1)
Length
50 mm
70 mm
100 mm
125 mm
NEW!
150 mm
250 mm
Guard
columns
NUCLEODUR Sphinx RP, 5 m

Particle size 5 m, pore size 110 ; special bifunctional RP phase, 14% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
Microbore columns
1 mm ID
761551.20
761551.30
761551.40
761551.46
761552.20
761552.30
761855.40
761855.46
761553.46
761554.20
761554.30
761554.40
761554.46
717682.10
717683.10
717684.10
760801.20
760801.30
760801.40
760801.46
760802.20
760802.30
760802.40
760802.46
760803.20
760803.30
760803.40
760803.46
761550.30
761550.30
761550.40
761550.40
2)
717680.10
EC columns 3)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
717681.10
760800.20
760800.30
760800.40
760800.46
761550.30
761550.30
761550.40
761550.40
Analytical columns with NUCLEODUR C18 ec

Particle size 3 m, pore size 110 ; octadecyl phase, endcapped, 17.5% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID 761001.20
3 mm ID 761001.30
4 mm ID 761001.40
4.6 mm ID 761001.46
Microbore columns
1 mm ID
761003.20
761003.30
761003.40
761003.46
761006.46
761004.20
761004.30
761004.40
761004.46
717711.10
717712.10
717713.10
760053.46
760052.20
760052.30
760052.40
760052.46
761005.30
761005.30
761005.40
761005.40
2)
717710.10
EC columns 3)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
NUCLEODUR
761002.20
761002.30
761002.40
761002.46
760050.20
760050.30
760050.40
760050.46
760051.20
760051.30
760051.40
760051.46
761005.30
761005.30
761005.40
761005.40
100-5 C18 ec
Particle size 5 m, pore size 110 ; octadecyl phase, endcapped, 17.5% C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID 761200.20
3 mm ID 761200.30
4 mm ID 761200.40
4.6 mm ID 761200.46
761150.20
761150.30
761150.40
761150.46
1 mm ID
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
717701.10
761350.20
761350.30
761350.40
761350.46
761380.46
761400.20
761400.30
761400.40
761400.46
717700.10
717702.10
717703.10
760008.46
760002.20
760002.30
760002.40
760002.46
761100.30
761100.30
761100.40
761100.40
3)
760004.20
760004.30
760004.40
760004.46
760001.20
760001.30
760001.40
760001.46
761100.30
761100.30
761100.40
761100.40

33
Analytical columns with NUCLEODUR C8 Gravity
30 mm 1)
Length
50 mm
70 mm
100 mm
125 mm
150 mm
250 mm
Guard
columns
Particle size 5 m, pore size 110 ; octyl phase, endcapped, 11 % C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
761755.20
761755.30
761755.40
761755.46
761751.20
761751.30
761751.40
761751.46
761752.46
761753.20
761753.30
761753.40
761753.46
761754.30
761754.30
761754.40
761754.40
760752.46
760753.20
760753.30
760753.40
760753.46
761754.30
761754.30
761754.40
761754.40
2)
760750.20
760750.30
760750.40
760750.46
760751.20
760751.30
760751.40
760751.46
Analytical columns with NUCLEODUR C8 ec

Particle size 3 m, pore size 110 ; octyl phase, endcapped, 10.5 % C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID 761018.20
3 mm ID 761018.30
4 mm ID 761018.40
4.6 mm ID 761018.46
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
761019.20
761019.30
761019.40
761019.46
761015.20
761015.30
761015.40
761015.46
761016.46
761017.20
761017.30
761017.40
761017.46
761012.30
761012.30
761012.40
761012.40
760061.46
760062.20
760062.30
760062.40
760062.46
761012.30
761012.30
761012.40
761012.40
2)
760063.20
760063.30
760063.40
760063.46
NUCLEODUR
760060.20
760060.30
760060.40
760060.46
100-5 C8 ec
Particle size 5 m, pore size 110 ; octyl phase, endcapped, 10.5 % C; eluent in column acetonitrile / water
ChromCart columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
EC columns
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
761705.20
761705.30
761705.40
761705.46
761701.20
761701.30
761701.40
761701.46
761702.46
761703.20
761703.30
761703.40
761703.46
761704.30
761704.30
761704.40
761704.40
760702.46
760703.20
760703.30
760703.40
760703.46
761704.30
761704.30
761704.40
761704.40
2)
760700.20
760700.30
760700.40
760700.46
760701.20
760701.30
760701.40
760701.46
Microbore columns and preparative columns with NUCLEODUR C8 phases are available on request!
34
Analytical columns with NUCLEODUR CN und CN-RP
Length
30 mm 1)
50 mm
125 mm
150 mm
250 mm
NUCLEODUR 100-3 CN-RP
Guard
columns
NEW!
Particle size 3 m, pore size 110 ; cyano phase (nitrile), 7% C; eluent in column acetonitrile / water
ChromCart columns
4.6 mm ID 761431.46
EC columns 2)
2 mm ID
3 mm ID
4 mm ID
4.6 mm ID
761430.40
760159.20
761430.30
761430.30
761430.40
761430.40
760157.30
760156.40
760156.46
NUCLEODUR 100-5 CN-RP

Particle size 5 m, pore size 110 ; cyano phase (nitrile), 7% C; eluent in column acetonitrile / water
ChromCart columns
4 mm ID
4.6 mm ID
761424.40
761424.46
EC columns 2)
4 mm ID
4.6 mm ID
760153.40
760153.46
760154.46
761423.40
761423.46
761420.40
761420.40
760152.40
760152.46
761420.40
761420.40
NUCLEODUR 100-5 CN
Particle size 5 m, pore size 110 ; cyano phase (nitrile), 7% C; eluent in column n-heptane
ChromCart columns
4 mm ID
4.6 mm ID
761422.40
761422.46
761421.40
761421.46
761419.40
761419.40
EC columns 2)
4 mm ID
4.6 mm ID
760151.40
760151.46
760150.40
760150.46
761419.40
761419.40
Microbore columns and preparative columns with these NUCLEODUR phases are available on request!
35
High performance HPLC packings

and
Applications
Nucleodur int2/5/0/10.2005 BD
Printed in Germany
www.mn-net.com
e-mail: Germany and international: sales-de@mn-net.com
USA:
sales-us@mn-net.com
Switzerland:
sales-ch@mn-net.com
France:
sales-fr@mn-net.com
MACHEREY-NAGEL
Germany: MACHEREY-NAGEL GmbH & Co. KG Neumann-Neander-Str. 6-8 D-52355 Dren Tel. +49 (0) 24 21 969-0 . Fax +49 (0) 24 21 969 199
USA:
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MACHEREY-NAGEL
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MN

HPLC Column c18 With Mobile Phase For Succinylsulfathizaole PG 18

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HPLC Column c18 With Mobile Phase For Succinylsulfathizaole PG 18

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Verfügbare Formate

High performance HPLC packings

DIN EN ISO 9001: 2000

NUCLEODUR Reversed Phase Packings

NUCLEODUR Particle Shape and Surface Symmetry

Fig. 1: High performance totally spherical NUCLEODUR silica

Fig. 2: Surface of a NUCLEODUR particle (magnified)

NUCLEODUR Purity Intolerant Towards Metals

NUCLEODUR 5 m measured by AAS

NUCLEODUR Reversed Phase Packings

125 x 4 mm NUCLEODUR 100-5 C18 ec

Fig. 3: Column stability of NUCLEODUR 100-5 C18 ec

back pressure [bar]

Surface area (BET)

Physical data of NUCLEODUR C18 ec

NUCLEODUR C18 Gravity

Fig. 5: Comparison of different base deactivated phases

Column: 250 x 4 mm NUCLEODUR C18 Gravity, 5 m

250 x 4 mm NUCLEODUR C18 Gravity, 5 m

Fig. 6: Separation of pyridine and phenol

NUCLEODUR C18 Gravity

Figure 8 shows the extent of protonation of surface

When is enhanced pH stability beneficial?

In figure 10 the excellent column stability of

after 5000 ml eluent

Column: 125 x 4 mm NUCLEODUR C18 Gravity, 5 m

Fig. 10: Stability of NUCLEODUR C18 Gravity at pH 1.5

NUCLEODUR C18 Gravity

Fig. 11 demonstrates the stability of NUCLEODUR

NUCLEODUR C18 Gravity

The pH stability of silica under alkaline conditions

125 x 4 mm NUCLEODUR C18 Gravity, 5 m

Fig. 13: Influence of the pH value on selectivity

NUCLEODUR C18 Pyramid

1) U. D. Neue et al., Chromatographia 54 (2001) 169 177

NUCLEODUR C18 Pyramid

Based on the ultrapure NUCLEODUR silica 3) the

Figure 15 shows the separation of various organic

Fig. 15: Separation of organic acids

NUCLEODUR C18 Pyramid

Column: NUCLEODUR C18

Fig. 16: Stability test comparison

NUCLEODUR C18 Pyramid

In addition to the exceptional polar selectivity

The perceptible increase in polarity has no impact

250 x 4 mm NUCLEODUR C18 Pyramid, 5 m

Fig. 17: Selectivity test

NUCLEODUR C18 Pyramid

Fig. 18: Tricyclic antidepressants

NUCLEODUR C18 Gravity

250 x 4 mm, 5 m particles

C18 or C8 the best of both worlds

In addition to the program of NUCLEODUR C18

NUCLEODUR 100-5 C18 ec

Eluents: A) water, B) methanol

Fig. 21: Phenols

250 x 4 mm NUCLEODUR 100-5 C

Indeed there are no general guidelines which could

Different from standard phenyl phases,

after 300 injections

50 x 4,6 mm NUCLEODUR Sphinx RP, 5 m

Fig. 22: Stability at pH 10

Due to the additional intermolecular interactions

Competitor 1 (column XP)