講稿方法轉(zhuǎn)換

UPLC

2

2

3

3

1

1

0

10

Minutes

20

0

2.6

Minutes

?2015WatersCorporation

30

5.2

Absorbance254nm

Absorbance254nm

UPLC

HPLC

….

,

UPLC?

HPLC

?2015WatersCorporation

?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:滯后體積包括進(jìn)樣器、阻尼器、混合器及管路

死體積/譜帶展寬體積

(

)

()

()

?2015WatersCorporation

14

?2015WatersCorporation 15

–

=1.04mL

–

ACQUITYUPLC?Hclass

=0.355mL

?2015WatersCorporation

16

?2015WatersCorporation

17

–

–

–

–

?2015WatersCorporation

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

第二代雜化顆粒技術(shù)允許HPLC方法首先轉(zhuǎn)化為XBridge?

柱的方法(基于選擇性考慮),然后再有效地轉(zhuǎn)換到UPLC?.

(lnamitriptyline/acenaphthene)

?2015WatersCorporation

21

–

–

?

?

–

?

?

?2015WatersCorporation

23

?2015WatersCorporation

24

–

–

?2015WatersCorporation

25

pH

?2015WatersCorporation

26

ACQUITYUPLC?

??O??

?2015WatersCorporation

27

20L進(jìn)樣量/2.49mL=0.8%

20L進(jìn)樣量/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

?2015WatersCorporation

32

從4.6x150mm到2.1x50mm,hS1.5mL/minlm

梯度

時(shí)間

流速

%A

%B

曲線

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

時(shí)間

流速

%A

%B

曲線

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