Basic Compounds, Sphingolipids, Antibiotics and Surfactants · (888) 456-HPLC LC-MS/MS Application...
Transcript of Basic Compounds, Sphingolipids, Antibiotics and Surfactants · (888) 456-HPLC LC-MS/MS Application...
[email protected](888) 456-HPLC
LC-MS/MS Application Tetracycline Antibiotics
Courtesy of Mr. N. MAEDA, ASSOCIATION OF MEAT SCIENCE & TECHNOLOGY INSTITUTE, JAPAN
Cadenza CW-C18, 100 x 2 mm + Guard column
A: 0.1% HCOOH aq., B: 0.1% HCOOH in ACN
5 - 95%B ( 0 - 7 min ), 95 - 5%B ( 7 - 8 min ), 5%B ( 8 - 17 min )
0.2 mL/min, 40 deg.C, 10 uL ( 20 ppb ), ESI, positive
OOOH OH
N
NH2
OH
OHO
OHHO
OOOH OH
N
NH2
OH
OHO
HO
OOOH OH
N
NH2
OH
OHO
HOCl
tetracycline(TC)
oxytetracycline(OTC)
chlortetracycline(CTC)
Time2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50%
0
100
2.50 5.00 7.50 10.00 12.50 15.00 17.50
%
0
100
20080509_OTC_02 MRM of 6 Channels ES+ TIC
3.04e47.29
7.07 7.98
20080509_OTC_02 MRM of 6 Channels ES+ 479.27 > 444
1.27e47.98
7.68
20080509_OTC_02 MRM of 6 Channels ES+ 479.27 > 154.1
8.82e37.98
20080509_OTC_02 MRM of 6 Channels ES+ 461.31 > 426.15
1.97e47.07
20080509_OTC_02 MRM of 6 Channels ES+ 461.31 > 201.2
2.52e37.07
20080509_OTC_02 MRM of 6 Channels ES+ 445.36 > 410.1
1.74e47.29
6.94
20080509_OTC_02 MRM of 6 Channels ES+ 445.36 > 154.1
1.20e47.33
OTC first ion
OTC second ion
TC first ion
TC second ion
CTC
CTC second ion
first ion
B a s i c C o m p o u n d s , S p h i n g o l i p i d s ,A n t i b i o t i c s a n d S u r f a c t a n t s
Improved peak shape for basic compoundsWide pore for small molecule analysis
1
0 5 10 15 min
water / acetonitrile = 45 / 5537 deg.C, 260 nm
Cadenza CW-C18, 150 x 2 mm0.6 mL/min, 25 MPa, N=16600
Cadenza CW-C18, 150 x 2 mm0.5 mL/min, 21 MPa, N=16900
Cadenza CW-C18, 150 x 2 mm0.4 mL/min, 17 MPa, N=17000
Cadenza CW-C18, 150 x 2 mm0.3 mL/min, 12 MPa, N=17200
12
34
5
Unison US-C18 (5um), 150 x 2 mm0.3 mL/min, 5 MPa, N=11200
4. naphthalene
3. toluene
CH3
2. benzene
5. fluorene
1. uracil
O
HO
A
B
C
D
E
High-Throughput performance with
wide-pore 3um particle ODS column
Cadenza CW-C18 has 1/3 surface area compared to normal pore-size column. The lower ODS coverage allows for half the analysis time under the same LC conditions. Cadenza CW-C18 is made with 3um high resolu-tion particle technology - producingincreased performance compared to 5um column technology (Chromatogram A
In addition, it can provideultra-high speed analysis with pressures that are acceptable for standard HPLC instruments (Chromatogram C to E).
Sub 2um particle columns require extremely high pressures. In contrast, Cadenza CW-C18 (3um) can be operated using normal (standard) HPLC condi-tions-- resulting in a balance of both low pressure and high plate counts. In addition, a 250mm length column (which is difficult to make using sub-2um particles) is available for high resolution and high speed analysis.
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3
4
mA
U
0 5 10 min
0
50
Water Soluble Vitamins
1
pyridoxine
N
CH2OHCH2OH
H3C
HO
2
HOCH2
H3C
H3C
CH
OH
C C
O
OHNHCH2CH2 C
O
(1/2Ca2)
calcium pantothenate
3
folic acid
HNCHC
ONH2N
CH2NHN
N
N C
C
O
CH2CH2OHOH
OH
O
4
riboflavin
N
NH
N
N
O
OH3C
H3C
CH2CH CHCH2OH
OH
CH
OH OH
Cadenza CW-C18, 150 x 3 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.07
3-30 %B (0-10min), 0.4 mL/min (9MPa), 37 deg.C
210 nm, 1 uL (0.1-1.2ug, ethanol, water)
Cadenza CW-C18 can provide many solutions to scientists working with basic compounds, high molecular weight componds, and hydrophilic compounds.
and B)
A wide-pore ODS column,
2
LC-MS/MS Application for Sphingolipids (Sphinganine-1-PO4)
Surfactants
XIC of -EMS: 379.4 to 383.4 amu from Sample 2 (dhS1P) of March_13_09_neg_EMS... Max. 4.6e6 cps.
1 2 3 4 5 6 7 8 9 10Time, min
0.0
5.0e5
1.0e6
1.5e6
2.0e6
2.5e6
3.0e6
3.5e6
4.0e6
4.5e64.69
-EMS: 4.535 to 4.860 min from Sample 2 (dhS1P) of March_13_09_neg_EMS_S1P....
376 378 380 382 384m/z, amu
2.0e4
4.0e4
6.0e4
8.0e4
1.0e5
1.2e5
1.4e5380.40
NH3
OH
+O P
O
O
OH
Sphinganine-1-Phosphate(Dihydrosphingosine-1-phosphate)
Cadenza CW-C18, 150 x 2 mmA: [water / HCOOH (100:0.2 v/v)] containing 7mM ammonium formateB: [methanol / HCOOH (100:0.2 v/v)] containing 5mM ammonium formate70-90%B (0-1min), 90%B (1-1.5min), 90-100%B (1.5-2.5min),100%B (2.5-6min), 100-40%B (6-6.5min), 40%B (6.5-10min)0.2mL/min - 0.28mL/min (0-6min), 0.2mL/min (6-10min)48deg.C, 5uL (50ug / mL)
Applied Biosystems QTrap, ESI Negative
Courtesy of Jennifer Ro and Joe WilliamsDepartment of Evolution, Ecology, and Organismal Biology,
Ohio State University - USA
1
2
mA
U
0 5 10 min
0
100
1 2
C8H17 OCH2CH2 nOH
n = ~10
Triton X-100
C9H19 OCH2CH2 nOH
n = ~12
Igepal CO-720
Cadenza CW-C18, 50 x 3 mmA: waterB: Methanol70-80 %B (0-6min)0.35 mL/min (4MPa), 37 deg.C, 260 nm2 uL (2% acetonitrile solution)
3
Internal DiameterPressure Limits of up to: 20MPa, 250 bar, 3000 psi
Column Length 1.0 mm 1.5 mm 2.0 mm 3.0 mm 4.6 mm 6.0 mm 10.0 mm10 CW020 CW030 CW00020 CW029 CW039 CW00930 CW011 CW071 CW021 CW031 CW001 CW061 CW0P150 CW012 CW072 CW022 CW032 CW002 CW062 CW0P275 CW013 CW073 CW023 CW033 CW003 CW063 CW0P3
100 CW014 CW074 CW024 CW034 CW004 CW064 CW0P4150 CW015 CW075 CW025 CW035 CW005 CW065 CW0P5250 CW016 CW076 CW026 CW036 CW006 CW066 CW0P6500 CW007
Guard Column System for Cadenza CW-C18Internal Diameter1.0 mm 1.5 mm 2.0 mm 3.0 mm 4.6 mm 6.0 mm 10.0 mm
Guard Holder GCH01S GCH01S GCH01S GCH01S GCH01S GCH01S GCH02MGuard Cartridge (Set of 3)
GCCW0C GCCW0C GCCW0S GCCW0S GCCW0S GCCW0S GCCW0M
All of our stationary phases can also be made in the following internal diameters: Nano: 0.05mm, 0.075mm Capillary: 0.1mm, 0.3mm, 0.5mm Semi-Prep: 20mm, 28mm
Ordering Information for Cadenza CW-C18 Columns 3µm
Polyethyline glycol (PEG)
polyethylene glycol #200025-30 %B (0-46 min)
2 uL (100 ug)
0 20 40
0
100
mV
min10 30 50 60
150
50
polyethylene glycol #400027-32 %B (0-46 min)
10 uL (500ug)
polyethylene glycol #600033-38 %B (0-46 min)
2 uL (100 ug)
Cadenza CW-C18 , 250 x 4.6 mmA: water B: acetonitrile1 mL/min (17-18 MPa), 37 deg.CELSD(spray chamber 50 deg.C, drift tube 100 deg.C)
Four Easy Ways To Order:1. Call us at (215) 665-8902
2. Order by fax (501) 646-34973. Through VWR (vendor code 8070779) or Fisher (vendor code VN101253)
4. Via www.imtaktusa.com with any major credit card