basic compounds, sphingolipids, antibiotics and surfactants · (888) 456-hplc lc-ms/ms application...

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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


1.97e47.07


2.52e37.07


1.74e47.29

6.94


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




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.

12

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)

[email protected]

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)

[email protected]

3

[email protected]

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


10 uL (500ug)


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

basic compounds, sphingolipids, antibiotics and surfactants · (888) 456-hplc lc-ms/ms application...

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