讲稿方法转换

UPLC

2

2

3

3

1

1

0

10

Minutes

20

0

2.6

Minutes

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30

5.2

Absorbance254nm

Absorbance254nm

UPLC

HPLC

….

,

UPLC™

HPLC

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©2015WatersCorporation

–

–

–

–

–

–

–

–

–

©2015WatersCorporation

–

–

–

–

©2015WatersCorporation

(min)

ml/min

%A

%B

0

1.5

95

5

-

2

15

1.5

5

95

6

3

20

1.5

5

95

1

4

30

1.5

95

5

1

–

–

2

3

–

–

–

–

–

1

0

10

20

30

Minutes

©2015WatersCorporation

Absorbance254nm

©2015WatersCorporation

10

©2015WatersCorporation

11

–

–

©2015WatersCorporation

12

–

–

–

©2015WatersCorporation

13

A

()

(

)

注意:滞后体积包括进样器、阻尼器、混合器及管路

死体积/谱带展宽体积

(

)

()

()

©2015WatersCorporation

14

©2015WatersCorporation 15

–

=1.04mL

–

ACQUITYUPLC®Hclass

=0.355mL

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16

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17

–

–

–

–

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18

©2015WatersCorporation

19

–

(k)

pka=9.4)

©2015WatersCorporation

20

3.6

3.3

WatersSpherisorb®S5P

2.7

2.4

2.1

1.8

1.5

1.2

0.9

0.6

0.3

WatersSpherisorb®S5CN

Nova-Pak®CNHP

Inertsil®Ph-3Hypersil®Phenyl

WatersSpherisorb®ODS1Resolve®C18

WatersSpherisorb®ODS2

Hypersil®CPSCyano

Inertsil®CN-3

YMC-Pack™Phenyl

µBondapak™C18

YMCJ'sphere™ODS–L80

Nova-Pak®Phenyl

Nucleosil®C18

Hypersil®BDSPhenyl

YMCbasic™

YMCJ'sphere™ODS–M80

XTerra®

ChromolithTMNova-Pak®

RP-18 C18

YMCJ'sphere™ODS–H80

YMC-Pack™CN

YMC-Pack™ProC4™

Luna™YMC-Pack™ODS–AQ™

YMC-Pack™ProC8™

C8

PhenylHexyl

Atlantis®dC18

ACTAce®C18Symmetry®C8

XTerraMSC8C8(2)

YMC-Pack™ProC18™

YMC-Pack™ODS-A™

Zorbax®XDBC18

Inertsil®ODS-3

SymmetryShield™RP8

XTerra®RP18

Luna®

C18Symmetry®C18

(2)

Zorbax®SBC18SymmetryShield™RP18

-0.3

-0.6

XTerra®RP8

ACQUITYUPLC™YMC-Pack™PolymerC18™BEH

-1.5

©2015WatersCorporation

-0.5

0.5

1.5

2.5

3.5

22

第二代杂化颗粒技术允许HPLC方法首先转化为XBridge™

柱的方法(基于选择性考虑),然后再有效地转换到UPLC™.

(lnamitriptyline/acenaphthene)

©2015WatersCorporation

21

–

–

•

•

–

•

•

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23

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24

–

–

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25

pH

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26

ACQUITYUPLC®

??O??

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27

20L进样量/2.49mL=0.8%

20L进样量/0.17mL=12%

©2015WatersCorporation

28

10µLx3.14x1.12x50

3.14x2.32x150

=

10µLx0.17

2.49

=10µLx0.068=0.7µL

©2015WatersCorporation

29

–

–

–

©2015WatersCorporation

30

©2015WatersCorporation 31

2.12

4.62

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32

从4.6x150mm到2.1x50mm,hS1.5mL/minlm

梯度

时间

流速

%A

%B

曲线

梯度段

(min)

梯度段

(cv)

Initial

0

1.5

95

5

*

0

0

2

15

1.5

5

95

6

15

3

20

1.5

5

95

1

5

4

30

1.5

95

5

1

10

=

x

=1.5mL/minx15min=

=xr2xL=3.14x2.32x150=2.49mL

©2015WatersCorporation

34

2.49mL =9.03cv

梯度

时间

流速

%A

%B

曲线

梯度段

(min)

梯度段

(cv)

Initial

0

1.5

95

5

*

0

0

2

15

1.5

5

95

6

15

9.03

3

20

1.5

5

95

1

5

3.01

4

30

1.5

95

5

1

10

6.02

©2015WatersCorporation 36

©2015WatersCorporation

37

©2015WatersCorporation

38

67

,-

%A

%B

;'

<(>)

<

(cv)

Initial

0

0.6

95

5

*

0

0

2

2.61

0.6

5

95

6

2.61

9.03

3

3.48

0.6

5

95

1

0.87

3.01

4

5.22

0.6

95

5

1

1.74

6.02

©2015WatersCorporation 40

ooo

o

©2015WatersCorporation

41

©2015WatersCorporation

42

2

3

HPLC4.6x150mm,5m

1/2=12

2/3=28

1

UPLC 2.1x50mm,1.7m

2

1

3

0

10

30

0

HPLC

10Minutes20

HClass

30

©2015WatersCorporation

44

Absorbance254nm

Absorbance254nm

–

–

–

–

–

–

©2015WatersCorporation

43

2

1

3

0

10

Minutes

20

30

©2015WatersCorporation

45

UPLC

2

3

1

0

2.6

Minutes

5.2

©2015WatersCorporation

46

Absorbance254nm

Absorbance254nm

©2015WatersCorporation

47

–

–

–

©2015W