Download Peptides Proteins Amino Acids

ON HPLC COLUMNS
Peptides
Proteins
Amino Acids
No.TI055E
LC and FAB Mass Analyses of D-Amino Acid-containing Peptides
LC
FAB-MS
LC
FAB-MS
[LC]
Cadenza CD-C18, 75 x 4.6 mm
5-45% of methanol/acetonitrile (6:4) in 50 mM phosphate, pH 5.7
1 mL/min
Courtesy of A. Sugihara, Osaka Municipal Technical Research Institute
TI055E-AI27
No.TI056E
Peptides - BSA Tryptic Digest
mAU
100
75
50
25
0
-25
-50
0
5
10
15
20
mAU
25
min
Brand-A ODS
100
75
50
25
0
-25
-50
0
5
10
15
20
25
min
mAU
Brand-B ODS
100
75
50
25
0
-25
-50
0
5
10
15
20
25
min
mAU
Brand-C ODS
100
75
50
25
0
-25
-50
0
5
10
15
20
25
min
150 x 2 mm
A: water / TFA = 100 / 0.08
B: acetonitrile / TFA = 100 / 0.05
5%B(0-2min), 5-40%B(2-20min), 40-90%B(20-25min), 90%B(25-30min), 90-5%B(30-33min)
0.2 mL/min
UV at 214 nm
40 °C
BSA Tryptic Digest, 18.5uL (ca.6.5ug)
TI056E-AJ10
No.TI057E
Peptides - BSA Tryptic Digest
mAU
100
80
60
40
20
0
-20
0
5
10
15
20
mAU
100
80
60
40
20
0
-20
25
min
Brand-A ODS
0
5
10
15
20
mAU
100
80
60
40
20
0
-20
25
min
Brand-B ODS
0
5
10
15
20
mAU
100
80
60
40
20
0
-20
25
min
Brand-C ODS
0
5
10
15
20
25
min
150 x 2 mm
A: acetonitrile / 1-propanol / water / acetic acid / TFA = 1 / 1 / 98 / 0.1 / 0.02
B: acetonitrile / 1-propanol / water / acetic acid / TFA = 70 / 20 / 10 / 0.09 / 0.02
5%B(0-2min), 5-40%B(2-20min), 40-90%B(20-25min), 90%B(25-30min), 90-5%B(30-33min)
0.2 mL/min
UV at 214 nm
40 °C
BSA Tryptic Digest, 18.5uL (ca.6.5ug)
TI057E-AJ10
No.TI076E
LC-MS Application of Myoglobin Tryptic Digest
mAU
UV at 215 nm
200
150
100
50
0
Ints.
BPC 300-1500 + MS
[MS]
Esquire 3000plus
( Ion Trap MS, positive mode )
5
4
3
2
1
0
0
5
10
15
20
25
30
35
40
45
Time [min]
Cadenza CD-C18, 150 x 2 mm
A: 0.01% TFA in water
B: 0.01% TFA in acetonitrile
2 - 50 %B ( 0 - 50 min )
0.2 mL/min, 45 °C
Myoglobin (Horse Heart), Tryptic digest
Courtesy of D.Higo, Bruker Daltonics K.K.
TI076E-BI10
No.TI081E
Amino Acids - 1
H O
H2N
1
HO
CH2
C
C
C
OH
2
O
L-aspartic acid (Asp / D)
CH2
C
C
3
OH
H2N
C
5
N
H
OH
L-serine (Ser / S)
C
CH2
C
C
OH
O
L-asparagine (Asn / N)
O
HO
OH
C
OH
6
H
L-hydroxyproline
50
H2N
C
H O
H2N
O
CH2
CH2
C
C
OH
L-glutamine (Gln / Q)
1
2
40
3
30
4
mAU
4
CH2
glycine (Gly / G)
H O
H2N
H2N
H O
H2N
O
20
5
6
10
0
0
5
10
15
20
25
30
min
Unison UK-C18, 250 x 4.6 mm
50mM H3PO4 + 10mM sodium heptanesulfonate
0.8 mL/min, 37 °C, 12.8 MPa, 210 nm, 2-10uL (4-20ug)
TI081E-BI13
No.TI082E
Amino Acids - 2
H O
H2N
1
CH
H3C
C
C
2
OH
H O
H2N
H3C
C
C
3
OH
H O
H2N
CH2
HS
C
C
OH
OH
L-threonine (Thr / T)
H2N
S
L-cysteine (Cys / C)
H O
CH2 C
C
H2N
H O
CH2 C
C
OH
5 HO
C
H O
H2N
O
CH2
CH2
C
C
OH
H3C
6
CH2
C
C
OH
H3C
OH
L-cystine
H O
H2N
L-glutamic acid (Glu / E)
L-valine (Val / V)
50
1
40
3
2
4
30
mAU
4
S
L-alanine (Ala / A)
20
5
10
6
0
0
5
10
15
min
20
25
30
35
Unison UK-C18, 250 x 4.6 mm
50mM H3PO4 + 10mM sodium heptanesulfonate / ACN = 92.5/7.5
0.8 mL/min, 37 °C, 13.5 MPa, 210 nm, 2-5uL (4-10ug)
TI082E-BI13
No.TI083E
Amino Acids - 3
CH2
C
H O
H2N
H O
H2N
1
C
2
OH
H3C
CH2
CH
C
C
OH
3
H O
H2N
H2N
CH2
CH2
CH2
CH2
C
C
H O
H2N
4
OH
HO
CH2
C
C
OH
CH3
L-phenylalanine (Phe / F)
H O
H2N
5
N
CH2
L-isoleucine (Ile / I)
C
C
O
N
H
CH2
CH2
CH2
C
C
OH
10
HN
CH2
C
C
8
OH
H O
H2N
H3C
S
CH2
C
C
OH
HO
3,4-dihydroxy L-phenylalanine
H O
H2N
C
HO
H
HN
CH2
CH2
CH2
C
C
OH
11
CH
CH2
C
C
H O
H2N
H O
H2N
H3C
L-methionine (Met / M)
OH
CH2
12
C
C
OH
H3C
H2N
N
L-ornithine
L-arginine (Arg / R)
L-leucine (Leu / L)
L-tryptophan (Trp / W)
1
140
4
2
120
5 3
6
100
7
80
mAU
H2N
N
H
7
L-tyrosine (Tyr / Y)
H O
H2N
OH
L-proline (Pro / P)
H O
H2N
C
6
OH
L-histidine (His / H)
9
L-lysine (Lys / K)
8 9
60
10
40
11
12
20
0
0
10
20
30
40
50
60
min
Unison UK-C18, 250 x 4.6 mm
50mM H3PO4 + 10mM sodium heptanesulfonate / ACN = 85 / 15
0.8 mL/min, 37 °C, 14.2 MPa, 210 nm, 0.4-10 uL (4-20 ug)
TI083E-BI17
No.TI097E
Peptides, Tryptic Digest of
-Casein
mAU
400
200
0
0
20
40
60
80
min
Cadenza CD-C18, 250 x 1 mm
A: 0.05% TFA in water
B: 0.05% TFA in acetonitrile
5 - 45 %B ( 0 - 80 min )
0.05 mL/min, 12 MPa, 37 °C, 210 nm
5 uL
TI097E-BL02
No.TI098E
Peptides, Tryptic Digest of
-Casein
200
mAU
150
100
50
0
0
5
10
15
20
min
Cadenza CD-C18, 50 x 2 mm
A: 0.05% TFA in water
B: 0.05% TFA in acetonitrile
5 - 35 %B ( 0 - 20 min )
0.5 mL/min, 10 MPa, 37 °C, 210 nm
5 uL
TI098E-BL02
No.TI100E
Peptides, Tryptic Digest of
-Casein
200
mAU
150
100
50
0
0
5
10
15
20
min
Unison UK-C18, 50 x 2 mm
A: 0.05% TFA in water
B: 0.05% TFA in acetonitrile
5 - 35 %B ( 0 - 20 min )
0.5 mL/min, 9 MPa, 37 °C, 210 nm
5 uL
TI100E-BL02
No.TI101E
Peptides, Tryptic Digest of
-Casein
150
mAU
100
50
0
0
20
40
60
80
min
Unison UK-C18, 250 x 2 mm
A: 0.05% TFA in water
B: 0.05% TFA in acetonitrile
5 - 45 %B ( 0 - 90 min )
0.2 mL/min, 14 MPa, 37 °C, 210 nm
5 uL
TI101E-BL02
No.TI153E
Peptides, Tryptic Digest of
-Casein
3 µm
Conventional ODS 5 µm
250 x 4.6 mm
500 x 4.6 mm
10-45%B ( 0-60min), 10µL
10-45%B ( 0-120min), 20µL
80 peaks
116 peaks
9.8 MPa
0
20
40
60
17.6 MPa
min 80
0
50
100
min
150
A: water / TFA, B:acetonitrile / TFA
0.5 mL/min, room temp., 220 nm
TI153E-DD01
No.TI170E
L-Carnitine
mAU
Unison UK-Silica
CH3
20
H3C
1
N+
CH3
H
CH2
C
OH
O
CH2
C
O-
L-carnitine
10
0
0
2
4
min 6
8
10
Unison UK-Silica, 150 x 3 mm
acetonitrile / 50mM ammonium acetate
= 65 / 35, 0.5 mL/min (7MPa)
37 °C, 210 nm, 2µL(20µg)
mAU
Unison UK-C18
1
20
10
0
0
2
4
min 6
8
10
Unison UK-C18, 75 x 4.6 mm
water / heptafluoro-n-butyric acid
= 100 / 0.1, 1 mL/min (7MPa)
37 °C, 210 nm, 2µL(20µg)
TI170E-DF09
No.TI188E
Theanine in Green Tea
mAU
100
CH3
CH2
NH
C
H O
H2N
O
CH2
CH2
C
C
OH
L-theanine
50
L-theanine, 2ug (2uL)
green tea, 20 µL
(theanine enriched)
0
0
5
10
15
min
Cadenza CL-C18, 250 x 3 mm
water / trifluoroacetic acid = 100 / 0.1
0.5 mL/min (15MPa), 37 deg.C, 210 nm
TI188E-DH20
No.TI191E
Betaine
mAU
CH3
50
betaine
H3C
N+
O
CH2
C
O-
CH3
0
0
5
10
min
Unison UK-Silica, 250 x 3 mm
acetonitrile / 50mM ammonium formate = 70 / 30
0.5 mL/min (10MPa), 37 deg.C, 210 nm, 2uL(20ug)
TI191E-DI01
No.TI282E
NBD-amino acids from rat plasma
A representative chromatogram of rat plasma amino acids derivatized with NBD-F.
1:NBD-Tau, 2:NBD-His, 3:NBD-Asn, 4:NBD-Gln, 5:NBD-Ser, 6:NBD-Arg, 7:NBD-Cit,
8:NBD-Asp, 9:NBD-Gly, 10:NBD-Glu, 11:NBD-Thr, 12:NBD-Ala, 13:NBD-Pro,
14:NBD-ε-amino-n-caproic acid (IS.), 15:NBD-Met, 16:NBD-Val, 17:NBD-Orn,
18:NBD-Lys, 19:NBD-Ile, 20:NBD-Leu, 21:NBD-Phe, 22:NBD-Tyr.
Cadenza CD-C18, 250 x 4.6 mm
A) water /acetonitrile /2-propanol /TFA = 90 /10 /0.8 /0.08
B) water /acetonitrile /2-propanol /TFA = 90 /10 /5 /0.08
C) water /acetonitrile = 90 /10
D) water /acetonitrile /TFA = 10 /90 /0.08
100%A, 0%BCD (0-25min), 100-0%A, 0-100%B (25-32min), 100%B (32-35min),
100-97%B, 0-3%D (35-39min), 97-0%B, 0-90%C, 3-10%D (39-45min),
90-65%C, 10-35%D (45-60min), 0-15%B, 65-55%C, 35-30%D (60-63min),
15%B, 55%C, 30%D (63-68min), 15%B, 55-50%C, 30-35%D (68-70min),
15%B, 50%C, 35%D (70-82min), 15-0%B, 50-40%C, 35-60%D (82-85min),
40%C, 60%D (85-100min), 100%A, 0%BCD (100-115min)
0.7 mL/min, 40 deg.C
Ex.470nm, Em.540nm
Ref.) M.Tomiya, T.Fukushima, J.Kawai, C.Aoyama, S.Mitsuhashi, T.Santa, K.Imai and T.Toyo'oka
Univ. of Shizuoka, Univ. of Tokyo and Musashino Univ.
Biomedical Chromatography, 20, 628-633 (2006)
Courtesy of Dr. Takeshi Fukushima, Univ. of Shizuoka
TI282E-FK08
No.TI291E
Peptides, Tryptic Digest of Hemoglobin (Human)
mAU
Cadenza CD-C18
5-30%B (0-115min)
70%B(115-120min)
158 peaks
40
Intrada WP-RP
0-30%B (0-115min)
70%B(115-120min)
80 peaks
0
0
50
100 min
250 x 3 mm
A: water /trifluoroacetic acid = 100 /0.1, B: acetonitrile /trifluoroacetic acid =100 /0.05
0.4 mL/min, 37 deg.C, 220 nm
TI291E-GA24
No.TI311E
Branched-chain Amino Acids
1
1
mV
200
H O
H2N
H3C
CH
CH2
C
C
OH
H3C
L-leucine
2
3
H3C
CH2
CH
C
CH3
L-isoleucine
100
3
H O
H2N
2
H3C
OH
H O
H2N
CH2
C
C
C
OH
H3C
L-valine
0
0
5
10 min
Unison UK-Amino, 150 x 2 mm
acetonitrile / 10mM ammonium acetate = 85 / 15
0.2 mL/min (4MPa), 37 deg.C, ELSD
0.5 uL (1.5ug)
TI311E-GD13
No.TI329E
Basic Amino Acids
O
34
1
mV
60
C
N
H
OH
H
L-proline
2
N
40
1
HN
3
C
C
OH
HN
(CH2)3
C
C
OH
unknown
L-arginine
4
15
H O
H2N
H2N
0
10
C
L-histidine
20
5
CH2
N
H
2
0
H O
H2N
20 min
H2N
H O
H2N
(CH2)4
C
C
OH
L-lysine
Unison UK-Amino, 250 x 4.6 mm
acetonitrile / 100mM ammonium formate = 76 / 24
1 mL/min (10MPa), 37 deg.C, ELSD, 3uL (1ug)
TI329E-GF14
No.TI371E
HPLC assay of methylated arginines using NBD-F
as a fluorescent reagent
NH2
HO
O
H
N
H
N CH
3
NH2
HO
N
H
O
CH3
N CH
H
N
NH2
HO
3
O
N
H
1: NG-monomethyl-L-arginine 2: NG,NG-dimethyl-L-arginine
(L-NMMA)
(ADMA)
(A) Standard mixture
H
N
NH2
N
CH3
H
N
HO
O
N
H CH3
3: NG,NG'-dimethyl-L-arginine
(SDMA)
H
N C H
3 7
N
H
4: N -propyl-L-arginine
(N-PLA)
(B) Rat plasma sample (200 uL) 4
10 uM
1
10 uM 10 uM 5 uM
2 3
4
1
2
3
min
min
Unison UK-C18, 150 x 4.6 mm
A: 50mM sodium phosphate buffer (pH 3.2) / acetonitrile = 91 / 9 (v/v)
B: acetonitrile
0%B (0-18 min), 0-30%B (18-28 min), 30%B (28-32 min)
0.75 mL/min, 40 deg.C, Ex.470 nm / Em. 530 nm
Pre-column derivatization: 105 ul of 100mM borate buffer (pH 9.0) and 30 uL of 40mM
NBD-F in acetonitrile were added to the 30 uL of sample. Then, the reaction was incubated
at 40 deg.C for 3 min. To stop the reaction, 435 uL of 0.5% acetic acid (v/v) was added.
A10 uL aliquot of sample was injected onto the HPLC.
Journal of Chromatography A, 1066 (2005) 41-45
Courtesy of Dr. TSUNODA, University of Tokyo, JAPAN
TI371E-GJ31
No.TI374E
Angiotensins
1. Angiotensin IV (Human)
5
2. Angiotensin III (Human)
4
3. Angiotensin I (Human)
3
4. [Val5]-Angiotensin I (Bovine)
2
5. Angiotensin II (Human)
mAU
1
normal phase
50
0
0
5
10
15 min
Unison UK-Amino
150 x 3 mm,
A: acetonitrile
B: 50mM ammonium acetate
10-35%B(0-15min)
0.4 mL/min 37 deg.C
220 nm, 2 uL (0.4ug)
mAU
2,5
1
50
4
3
0
0
5
10
15 min
reversed phase
Unison UK-C18
150 x 3 mm
A: water/ TFA = 100/ 0.1
B: acetonitrile/ TFA = 100/ 0.1
20-30%B(0-15min)
0.4 mL/min, 37 deg.C
220 nm, 2 uL (0.4ug)
TI374E-GK02
No.TI377E
Betaine, Carnitine, GABA
1
CH3
H3C
N+
O
CH2
C
2
CH2
H3C
N+
O-
CH3
H
CH2
CH2
O
C
H2N
OH
O-
OH
CH2
betaine
C
3
O
L-carnitine
GABA
1
mV
2
50
3
0
0
2
4
min
Unison UK-Amino, 50 x 3 mm
A: acetonitrile
B: 10mM ammonium acetate
15-30%B(0-5min)
0.5 mL/min, 37 deg.C, ELSD, 1 uL (1ug)
TI377E-GK26
No.TI387E
Calcitonin in Gelatin Aqueous Solution
gelatin
calcitonin
mAU
calcitonin acetate, salmon (MW 3492)
Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-
100
Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-LysLeu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-ThrGly-Ser-Gly-Thr-Pro-NH2
.CH3COOH
50
calcitonin in gelatin aqueous solution
calcitonin in water
gelatin in water
0
0
1
2
3
4 min
Cadenza HS-C18, 30 x 3mm
A: water /trifluoroacetic acid = 100 /0.1
B: acetonitrile /trifluoroacetic acid = 100 /0.07
0-45%B (0-2min), 0.4mL/min (3MPa), 37deg.C, 220nm
2uL (calcitonin: 1ug, gelatin: 10ug)
TI387E-HB01
No.TI388E
Calcitonin, Salmon
calcitonin acetate, salmon (MW 3492)
mV
Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-
50
Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-LysLeu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-ThrGly-Ser-Gly-Thr-Pro-NH2
.CH3COOH
normal phase
0
2
4
6 min
mAU
0
Unison UK-Amino, 100 x 3 mm
A: acetonitrile
B: 50mM ammonium acetate
5-100%B(0-5min)
0.4mL/min (2MPa), 37deg.C, ELSD
1.5uL (1.5ug)
150
Cadenza CW-C18
100
Cadenza CL-C18
reversed phase
50
Cadenza CD-C18
Unison UK-C18
0
0
2
4
6 min
100 x 3 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
30-50%B(0-5min)
0.4mL/min (4MPa), 37deg.C, 220nm
1uL (1ug)
TI388E-HB05
No.TI393E
Glutathiones
1
2 HO
H NH2
H
N
HO
O
O
HS
O
O
H
H NH2
N
H
O
H
N
O
H
N
H
OH
O
S
OH
S
O
O
HO
H NH2
glutathione reduced (GSH)
O
N
H
H
H
N
O
OH
O
glutathione oxidized (GSSG)
1
mAU
2
50
0
0
2
4
6
8 min
UK-Amino, 100 x 4.6 mm
A: acetonitrile, B: 50mM ammonium acetate
80-100 %B (0-8min), 1 mL/min
37 deg.C, 220 nm, 5 uL(5ug)
TI393E-HB14
No.TI419E
Microcystins
O
H
N
OH
O
O
O
N
H HN
N
NH H
NH2
NH
OCH3
1
O
HN
= H N
2
O
CH2 O
NH
NH
N
O
O
N
H
(CH2)3
microcystin RR
OH
3
2
CH2
=
2
HO
1
microcystin YR
3
mAU
2
3
1
H 3C
(CH2)2
=
40
2
3
H 3C
microcystin LR
1
3
2
1
3
20
2
1
3
2
1
0
0
5
10
min
150 x 3 mm,
A: water / TFA = 100 / 0.05
B: acetonitrile / TFA = 100 / 0.05
35-60 %B (0-10min)
0.4 mL/min, 37 deg.C
220 nm, 5 uL (5 ppm)
TI419E-HG01
No.TI420E
Microcystins
O
H
N
OH
O
O
O
N
H HN
N
NH H
1
NH2
NH
NH
= H N
2
OCH3
N
H
(CH2)3
microcystin RR
O
HN
NH
N
O
2
O
CH2 O
O
H 3C
(CH2)2
=
microcystin LR
H 3C
OH
3
CH2
=
mAU
2
microcystin YR
HO
3
50
1
0
0
5
10
min
Unison UK-Amino, 150 x 3 mm
A: acetonitrile, B: 50mM ammonium acetate, 20-30 %B (0-10min)
0.4 mL/min (4MPa), 37 deg.C, 220 nm, 5 uL (50 ppm)
TI420E-HG01
No.TI423E
NDA-amino acids
CHO
CHO
NDA (naphthalene-2,3-dicarboxaldehyde)
Relative Fluorescence(mV)
700
600
Asp
Glu
Gly
Asn His Ser
Arg
500
400
Ala
GABA
Tau
Tyr
Gln
300
Thr
200
100
0
-100
0
5
10
15
20
25
30
Time (min)
Cadenza CD-C18, 150 x 2 mm
A: 50mM phosphate buffer (pH 5.8) /acetonitrile /methanol = 15 /2 /3
B: 50mM phosphate buffer (pH 5.8) /acetonitrile /THF = 73 /18 /9
0-100%B (0-30min), 0.3 mL/min, 37 deg.C, FLS (Ex.420nm, Em.480nm)
standard amino acids (10 pmol) derivatized with NDA
TI423E-HG08
No.TI425E
Microcystins
O
H
N
OH
O
O
N
NH H
N
H HN
O
NH2
1
NH
NH
= H N
2
OCH3
O
HN
(CH2)3
microcystin RR
O
CH2 O
N
H
NH
N
O
mAU
O
2
OH
CH2
=
HO
microcystin YR
10
3
H 3C
(CH2)2
=
H 3C
microcystin LR
3
20 ppb x 100 uL
2
1
3
5
2
1
40 ppb x 50 uL
3
100 ppb x 20 uL
2
1
3
2
1 ppm x 2 uL
1
0
0
1
2
3
4
min
Unison US-C18, 50 x 2 mm
A: water / TFA = 100 / 0.05
B: ACN / TFA = 100 / 0.05
30-40 %B (0-13min)
0.4 mL/min
37 deg.C, 240 nm
TI425E-HH06
No.TI469E
Aliphatic Amino Acids
mAU
1
6
CH2
CH
H 3C
5
H O
H 2N
H 3C
C
C
L-leucine (Leu / L)
40
H O
H 2N
2
OH
H 3C
CH
CH2
C
C
OH
CH3
L-isoleucine (Ile / I)
O
2
C
3
N
H
OH
H
L-proline (Pro / P)
3
4
4
H O
H 2N
H 3C
CH2
C
C
H 3C
L-valine (Val / V)
20
1
H O
H 2N
5
H 3C
OH
C
C
OH
L-alanine (Ala / A)
O
6
H 2N
CH2
C
OH
glycine (Gly / G)
0
0
10
20
min
Unison UK-Amino, 250 x 2 mm
A: acetonitrile
B: 10mM ammonium acetate
10-30 %B (0-20min)
0.16 mL/min (5MPa)
37 deg.C, ELSD, 1 uL (0.5 ug)
TI469E-HL26
No.TI470E
Hydrophilic Amino Acids
mAU
4
H O
H 2N
1 H 3C
3
CH
C
C
OH
OH
L-threonine (Thr / T)
20
O
2
H 2N
CH2
C
OH
glycine (Gly / G)
C
3 H 2N
2
CH2
CH2
4 H 2N
C
CH2
O
C
OH
H O
5
CH2
OH
0
20
min
C
OH
H O
H 2N
1
C
L-asparagine (Asn / N)
10
10
C
L-glutamine (Gln / Q)
5
H 2N
0
H O
H 2N
O
C
C
OH
L-serine (Ser / S)
Unison UK-Amino, 250 x 2 mm
A: acetonitrile
B: water /acetic acid = 100 /1
20-25 %B (0-20min)
0.2 mL/min (8MPa)
37 deg.C, ELSD, 0.4 uL (0.4 ug)
TI470E-HL26
No.TI471E
Acidic Amino Acids
mAU
1
HO
C
1
H O
H 2N
O
CH2
CH2
C
C
OH
L-glutamic acid (Glu / E)
100
2
HO
0
0
5
C
O
2
min
H O
H 2N
CH2
C
C
OH
L-aspartic acid (Asp / D)
Unison UK-Amino, 50 x 2 mm
A: acetonitrile
B: water /acetic acid = 100 /1
70-90 %B (0-5min)
0.2 mL/min (3MPa)
37 deg.C, ELSD, 1 uL (1ug)
mAU
1
50
2
0
0
5
10
15
20
min
Unison UK-Amino, 250 x 2 mm
A: acetonitrile
B: water /acetic acid = 100 /1
70-90 %B (0-20min)
0.2 mL/min (16MPa)
37 deg.C, ELSD, 2 uL (2ug)
TI471E-HL26
No.TI472E
Sulfur-containing Amino Acids
1
H O
H 2N
1
H 3C
S
CH2
C
C
OH
L-methionine (Met / M)
2
3
H O
H 2N
HS
CH2
C
C
OH
mAU
L-cysteine (Cys / C)
50
3
Cl-
H 2N
S
S
H O
CH2 C
C
H 2N
H O
CH2 C
C
OH
OH
L-cystine
2
0
0
10
20
min
Unison UK-Amino, 250 x 2 mm
A: acetonitrile
B: 50mM ammonium acetate
50-90 %B (0-20min)
0.2 mL/min (7MPa)
37 deg.C, ELSD, 5 uL (2-5ug)
TI472E-HL26
No.TI473E
Aromatic Amino Acids
2
1
H O
H 2N
CH2
C
C
OH
L-phenylalanine (Phe / F)
3
2
H O
H 2N
mAU
CH2
N
C
C
OH
L-tryptophan (Trp / W)
20
3
H O
H 2N
HO
CH2
C
C
OH
L-tyrosine (Tyr / Y)
1
0
0
10
20
min
Unison UK-Amino, 250 x 2 mm
A: acetonitrile
B: 10mM ammonium acetate
20-50 %B (0-20min)
0.2 mL/min (6MPa)
37 deg.C, ELSD, 2 uL (1 ug)
TI473E-HL26
No.TI474E
Glu
Asp
His
Thr
Tyr
Ala
Met
Val
Pro
20
Cys
Gly
Phe
Leu
Ile
Trp
40
Gln
mV
Arg, Ser, Lys
Asn
Normal Phase Separation for Amino Acids
0
20
min
Unison UK-Amino, 100 x 3 mm
A: ACN*1 /THF*2 /50mM TFAA*3
= 46 /46 /8
B: MeOH*4 /50mM HCOONH4
/50mM TFAA = 20 /20 /60
0-5 %B(0-20min)
5 %B(20-25min)
30-60 %B(25-40min)
0.5 mL/min, 37 deg.C, ELSD
2 uL(1-2ug)
Cys
Glu
Asp
Lys
Thr
Ala
Arg
Val
Asn
Pro
Tyr
Phe
Ile Leu
mV
50
30
0
0
10
*1 : acetonitrile
*2 : tetrahydrofuran
*3 : ammonium trifluoroacetate
*4 : methanol
Gly, Ser, His, Gln
10
Trp
Met
0
Unison UK-Amino, 100 x 3 mm
A: ACN*1 /THF*2 /50mM TFAA*3
= 46 /46 /8
B: MeOH*4 /50mM HCOONH4
= 20 /80
0-5 %B(0-15min)
8 %B(15-25min)
8-60 %B(25-35min)
0.4 mL/min, 37 deg.C, ELSD
2 uL(1-4ug)
20
30
40 min
*1 : acetonitrile
*2 : tetrahydrofuran
*3 : ammonium trifluoroacetate
*4 : methanol
TI474E-IA06
No.TI521E
Glutathiones
1
H NH2
2
mV
HO
100
O
O
H
N
O
HS
H
N
H
OH
O
glutathione reduced (GSH)
1
2
H NH2
HO
O
O
H
N
O
H
N
H
OH
O
S
S
O
50
HO
H NH2
O
N
H
H
H
N
O
OH
O
glutathione oxidized (GSSG)
0
0
2
4
Imtakt USA ([email protected])
North America
6 min
50 x 3 mm
A: 5mM ammonium acetate
B: 100mM ammonium acetate / ACN
= 50 / 50
0-100 %B (0-5min), 0 %B (5-10min)
0.4 mL/min (3MPa), 37 deg.C, ELSD
2 uL (2ug)
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI521E-IF17
No.TI524E
Anserine and related compounds
1
mV
1,3
H 2N
O
OH
O
1
3
K+ 2
200
H
N
N
N
H
L- carnosine
(beta-alanyl-L-histidine)
H
N
H 2N
O
OH
O
2
N
.HNO3
N
100
CH3
L- anserine nitrate
(beta-alanyl-1-methyl-L
-histidine nitrate)
K+
2
O
O
3
KO
N
HN
P OCH2
N
O
N
OK
0
HO
0
5
10 min
OH
Inosine 5'-monophosphate
dipotassium
150 x 3 mm
A: 10mM ammonium acetate
B: 50mM ammonium acetate / acetonitrile = 80 / 20
0-100%B (0-10min), 0.4 mL/min (9MPa), 37deg.C
ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
5 uL (2-6ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI524E-IF22
No.TI526E
Lys
Arg
His
basic amino acids
0-10 %B (0-15min)
Met
Trp
hydrophilic amino acids
0-10 %B (0-15min)
Phe
Tyr
400
Gln
mV
Gly, Ala, Pro, Thr, Ser, Asn, Asp, Glu, Cys
Amino acids: neutral pH condition
200
Ile
Leu
Val
aliphatic amino acids
0-20 %B (0-15min)
branched-chain amino acids (BCAA)
0-10 %B (0-15min)
0-10 %B (0-15min)
0
0
5
10
15
min
Scherzo SM-C18, 150 x 3 mm
A: 5mM ammonium acetate
B: 125mM ammonium acetate / acetonitrile = 20 / 80
0.4 mL/min (8MPa), 37 deg.C
ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
2-5 uL (2.5-4ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI526E-IG22
No.TI527E
Asp
Glu
Gln
Trp
Phe
Tyr
hydrophilic amino acids
0-10 %B (0-10min)
aliphatic amino acids
0-30 %B (0-10min)
Ile
Leu
200
acidic amino acids
0-30 %B (0-10min)
Met
400
Val
mV
Gly, Ala, Pro, Thr, Ser, Asn, His, Lys, Arg, Cys
Amino acids: acidic condition
branched-chain amino acids (BCAA)
0-10 %B (0-10min)
0-10 %B (0-10min)
0
0
5
10
min
Scherzo SM-C18, 150 x 3 mm
A: water / acetic acid = 100 / 0.1
B: acetonitrile / acetic acid = 100 / 1
0.4 mL/min (8MPa), 37 deg.C,
ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
2-5 uL (1-5ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI527E-IG22
No.TI528E
Amino acids: normal phase
Gly
Lys
Leu
Ile
Val
mV
Tyr
Phe
500
Met
Asn
His
Thr
Glu
Asp
Arg
Cys
Pro
Gln
Trp
Ala
Ser
0
0
5
10
Imtakt USA ([email protected])
North America
15
min
Scherzo SM-C18
150 x 3 mm
A: acetonitrile / acetic acid
= 100 / 0.5
B: water / acetic acid
= 100 / 0.5
2-50 %B(0-15min)
0.4 mL/min (3MPa)
37 deg.C,
ELSD
(spray chamber 50 deg.C,
drift tube 100 deg.C)
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI528E-JB15
No.TI597E
Trp
Ile
Leu
Met
Car
Pro
Arg
Glu
Tau
Gly
Ser
Ala
Lys
Asp
Val
each 100 µg/mL
Phe
RP analysis of intact amino acids using Corona CAD
each 1 µg/mL
0
5
10
15
20 min
Cadenza CD-C18, 250 x 4.6 mm
A: 0.15% TFA
B: 0.15% TFA in acetonitrile
0%B (0-5min), 0-40%B (5-20min), 40%B (20-23min)
40-0%B (23-23.1min), 0%B (23.1-35min)
0.6 mL/min, 25 °C
Corona CAD ( GAS=35psi, Filter: None )
10 µL
Courtesy of Kyoko Fukushima, LMS CO., LTD. / JAPAN
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI597E-JC30
No.TI605E
Peptide mapping
Tryptic digest of alpha-casein
mAU
320 peaks
1-50 %B (0-150min)
0.2 mL/min (25 MPa)
100
50
198 peaks
5-37 %B (0-150min)
0.4 mL/min (13 MPa)
0
0
50
100
150
min
250 x 3 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
50 deg.C, 220 nm, 5 uL
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI605E-JD14
No.TI646E
Peptide mapping (50°C, formic acid)
mV
15
Presto FF-C18
0.4 mL/min(22MPa)
10
5
Cadenza CD-C18
1 mL/min(12MPa)
0
0
20
40
60
min
250 x 4.6 mm
A: water / formic acid = 100 / 0.5
B: acetonitrile / formic acid = 100 / 0.5
2-50 %B (0-60min)
50 deg.C, ELSD(spray chamber 35 deg.C, drift tube 100 deg.C)
tryptic digest of alpha-casein 20 uL
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI646E-KC14
No.TI653E
The effect of TFA modifier on peptide mapping
mV
20
A: water /formic acid /TFA
= 100 /0.5 /0.02
B: acetonitrile /formic acid /TFA
= 100 /0.5 /0.02
2-50 %B (0-60min)
10
A: water /formic acid
= 100 /0.5
B: acetonitrile /formic acid
= 100 /0.5
2-50 %B (0-60min)
0
0
20
40
60 min
Presto FF-C18, 250 x 4.6 mm
0.4 mL/min (22MPa), 50 deg.C
ELSD(spray chamber 35 deg.C, drift tube 100 deg.C)
tryptic digest of alpha-casein 20 uL
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI653E-KD05
No.TI660E
Collagenpeptide
collagenpeptide, enzyme hydrolyzed
mV
50
collagenpeptide, acid hydrolyzed
0
0
50
100 min
Presto FF-C18, 250 x 4.6 mm
A: water / formic acid / TFA = 100 / 0.5 / 0.02
B: acetonitrile / formic acid / TFA = 100 / 0.5 / 0.02
1-50 %B (0-90min)
0.4 mL/min (21MPa), 50 deg.C
ELSD (spray chamber 35deg.C, drift tube 100deg.C)
20 uL (20ug, water)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI660E-KD20
No.TI682E
Histamine related compounds
mAU
O
N
3
1
1
100
NH2
N
H
L-histidine
OH
N
4
2
OH
2
O
N
CH3
N-methyl-4imidazoleacetic acid
5
50
N
3
O
N
H
4-imidazoleacetic acid
N
4
0
0
10
20
NH2
N
CH3
N-methylhistamine
min
N
5
OH
NH2
N
H
histamine
Scherzo SS-C18, 150 x 3 mm
A: methanol
B: 100 mM ammnonium formate : 100 mM ammonium hydroxide = 95 : 5 (pH8.1)
0-20 %B (0-20 min)
0.4 mL/min (6 MPa), 60 deg.C, 230 nm, 1.6 uL ( 0.6-2.8 ug, MeOH)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI682E-KH09
No.TI683E
Serotonin related compounds
1
HO
mAU
O
3
1
NH2
N
H
5-hydroxy-L-tryptophan
4
300
OH
2
15-50 %B (0-15 min)
9 MPa
HO
NH2
2
1
200
N
H
serotonin
3
2
4
10-55 %B (0-15 min)
9 MPa
O
3
NH2
N
H
L-tryptophan
1
100
OH
3
4
HO
2
25-90 %B (0-15 min)
12 MPa
0
OH
4
N
H
5-hydroxyindole
O
-3-acetic acid (5-HIAA)
0
5
10
15 min
150 x 3 mm
A: water / formic acid = 100 / 0.1
B: 100 mM ammnonium formate / acetonitrile = 70/ 30
0.4 mL/min, 37 deg.C, 260 nm, 1.2 uL ( 0.4 ug, 0.1% formic acid)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI683E-KH10
No.TI685E
Peptide mapping using LC-FTMS
RT: 4.9 - 50.2
748.37
100
634.36
0-40%B (0-60 min)
0.2 mL/min
90
80
Relative Abundance
70
523.22
60
598.34
491.29
50
457.21
412.72
880.48
940.48
1199.61
669.34
1053.49
40
30
20
10
796.42
416.75
486.25
742.45
755.87
667.34
415.73
416.20
905.48
473.77
575.32
801.73 650.36
821.39
787.94
1052.54
473.76 416.20
523.22
416.20
692.87
1081.02 7
0
748.37
100
0-60%B (0-60 min)
0.7 mL/min
90
Relative Abundance
80
634.35
70
60
880.48
416.19
412.72
669.34
40
30
1053.48
598.34
50
455.74
1199.61
491.29
457.21
494.31
905.47
415.73
486.25
20
623.82
10
495.75
755.86
473.77
742.45
787.94
692.87
643.31
1081.02
684.20 684.20 684.20
480.29
684.20
0
6
8
10
12
14
16
18
20
22
24
26
28
Time (min)
30
32
34
36
38
40
42
44
46
48
Presto FF-C18, Cadenza CD-C18, 150 x 3 mm
A: water / formic acid / TFA = 100 / 0.5 / 0.02
B: acetonitrile / formic acid /TFA = 100 / 0.5 / 0.02
50 deg.C, 2uL (tryptic digest of alpha-casein)
FTMS (Thermo Scientific Exactive), ESI-positive (Full ms)
Courtesy of Dr. KUWATA Keiko, Kyoto University, JAPAN
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI685E-KH12
No.TI262E
Interferon
mAU
unknown
interferon alfa-2b
(genetical recombination, 19,269 Da)
40
0
0.5
1.0 min
mAU
unknown
0
Intrada WP-RP, 30 x 2mm
A: water / TFA = 100 / 0.1
B: acetonitrile / TFA = 100 / 0.07
20-45%B (0-0.5min), 45%B (0.5-1min)
0.8 mL/min(10 MPa), 37deg.C, 280nm
15uL (180ng)
20
interferon alfa-2b
(genetical recombination, 19,269 Da)
0
0
5
10
15min
Intrada WP-RP, 250 x 2mm
A: water / TFA = 100 / 0.1
B: acetonitrile / TFA = 100 / 0.07
15-50%B (0-9min), 50%B (9-12min)
0.2 mL/min(13 MPa), 37deg.C, 280nm
10uL (120ng)
TI262E-FA23
No.TI263E
Interferon
mAU
interferon alfa-2b
(genetical recombination, 19,269 Da)
unknown
40
0
0
5
10
15 min
Intrada WP-RP, 250 x 4.6 mm
A: water / TFA = 100 / 0.1
B: acetonitrile / TFA = 100 / 0.07
15-50%B (0-9min), 50%B (9-12min)
1 mL/min (14 MPa),37deg.C, 220nm
8uL (72ng)
TI263E-FA23
No.TI270E
Protein, beta-Glucosidase
mAU
bata-glucosidase
(almond, 183 kDa)
40
20
0
0
5
10
15
20 min
Intrada WP- RP, 250 x 4.6 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
30-50%B (0-15min)
1.0 mL/min (16 MPa)
37 deg.C, 280 nm, 8 uL (42.4 ug)
TI270E-FG04
No.TI271E
mAU
Protein, Thyroglobulin
40
thyroglobulin
(porcin thyroid, 670 kDa)
20
0
0
5
10
15
20 min
Intrada WP- RP, 250 x 4.6 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
10-95%B (0-15min)
1.0 mL/min (16 MPa)
37 deg.C, 280 nm, 8 uL (40 ug)
TI271E-FG04
No.TI272E
Protein, Hemoglobin
mAU
hemoglobin
(bovin blood, 64 kDa)
40
20
0
0
5
10
15
20 min
Intrada WP- RP, 250 x 4.6 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
20-65%B (0-15min)
1.0 mL/min (16 MPa)
37 deg.C, 280 nm, 8 uL (42.4 ug)
TI272E-FG04
No.TI273E
Protein, Lactferrin
mAU
lactferrin
(bovin milk, 93 kDa)
40
20
0
0
5
10
15 min
Intrada WP- RP, 250 x 4.6 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
20-50%B (0-15min)
1.0 mL/min (16 MPa)
37 deg.C, 280 nm, 3 uL (15.3ug)
TI273E-FG04
No.TI274E
Bovin Serum
20
30
mAU
10
100
0
0
10
20
30
40min
Intrada WP- RP, 250 x 4.6 mm
A: water /TFA = 100 /0.1
B: acetonitrile /TFA = 100 /0.07
25-50%B (0-40min)
1.0 mL/min (16 MPa)
37 deg.C, 220 nm, 0.4 uL
TI274E-FG04
No.TI293E
mAU
Bovine Milk
500
220 nm
280 nm
0
0
10
20
30 min
Intrada WP-RP, 250 x 3 mm
A: water /trifluoroacetic acid = 100 /0.1,
B: acetonitrile /trifluoroacetic acid =100 /0.05
0-100%B (0-25min), 100%B(25-30min)
0.4 mL/min (12MPa), 37 deg.C, 5uL
TI293E-GB01
No.TI430E
IgG2 mAb by reversed-phase analysis
Intact IgG2 mAb
280 nm
Reduced IgG2 mAb
215 nm
Ref.) T. M. Dillon ., P.V. Bondarenko, D.S. Rehder, G.D. Pipes, G. R. Kleemann, M.S. Ricci
Journal of Chromatography A, 1120 (2006) 112-120
Intrada WP-RP, 150 x 2 mm
A: 0.1% TFA, B: IPA /ACN /water /TFA = 70 /20 /9.9 /0.1
10%B (0-2 min), 25%B (2-4 min), 25-32%B (4-25 min)
0.5 mL/min, 75 deg.C, 280 nm (upper) or 215nm (lower)
Courtesy of Dr.Thomas Dillon, AMGEN Inc., USA
TI430E-HI27
No.TI440E
A proteomics study on human breast cancer cell lines
by fluorogenic derivatization-liquid chromatography/tandem
mass spectrometry
2
9
6
1
DAABD-proteins
11
4
3
13
12
5
7
8
10
Breast cancer cell _ 1
Breast cancer cell _ 2
Breast cancer cell _ 3
Breast cancer cell _ 4
Breast cancer cell _ 5
Breast cancer cell _ 6
Breast cancer cell _ 7
Normal mammary epitherial cell
Ref-1: Biomedical Chromatography, 2008, 22 (11), 1304-1314
Ref-2: Journal of Proteome Research, 2007, 6, 2841-2849
Intrada WP-RP, 250 x 4.6 mm
A: 0.15% TFA in (acetonitrile /isopropanol /water = 9 /1 /90)
B: 0.15% TFA in (acetonitrile /isopropanol /water = 69 /1 /30)
C: 0.20% TFA in (acetonitrile /isopropanol /water = 9 /1 /90)
5%B,1%C(0-5min), 5-30%B,1-35%C (5-30min), 30%B,35%C(30-65min)
30-35%B,35%C(65-70min), 35-38%B,35%C(70-130min)
38-44%B,35-55%C(130-250min), 44%B,55%C(250-300min)
44-47%B,55-53%B(300-330min), 47-60%B,53-40%C(330-480min)
60-70%B,40-30%C(480-520min), 70-90%B,30-10%C(520-570min)
0.55 mL/min, 60 deg.C, FLD( Ex.395nm, Em.505nm)
Courtesy of Kazuhiro Imai, Ph.D. Dhc., Professor & Dean, Musashino University, Japan
TI440E-HJ15
No.TI289E
Proteins
thyroglobulin (porcin thyroid, 670 kDa, pI=4.58)
xanthine oxidase (buttermilk, 360 kDa)
beta-glucosidase (almond, 183 kDa)
lactoferrin (bovine milk, 80 kDa)
hemoglobin (Bovine, 66-68 kDa, pI=6.8)
albumin (human serum, 66.4 kDa, pI=4.7)
albumin (bovine, 66.3 kDa, pI=4.9)
hemoglobin (Human, 64.5 kDa, pI=6.8)
albumin (eggs, 45 kDa, pI=4.54-4.9)
deoxyribonuclease I (bovine pancreas, 31 kDa, pI=4.78-5.2)
alpha-chymotryosinogen A (bovine pancreas, 25.7 kDa, pI=8.97)
papain (papaya latex, 21 kDa, pI=8.75-9.55)
lysozyme (chicken egg white, 14 kDa, pI=11.0)
alpha-lactalbumin (bovine milk, 14.2 kDa, pI=4.5)
cytochrome c (horse heart, 12.4 kDa, pI=9.1)
insulin (bovine pacreas, 5733 Da)
angiotensin II (1046 Da)
0
5
10
15 min
Intrada WP-RP, 150 x 4.6 mm, 1 mL/min, 37 degC, 280 nm
A: water /TFA=100 /0.1, B: ACN /TFA=100 /0.1, 20-70%B (0-15min)
TI289E-GA16
No.TI560E
Polyglutamic acid
Na+
mV
MW: 1,500,000 - 2,500,000
400
O
HN
CH
HO
C
CH2
CH2
C
O
Na+
n
poly-gamma-glutamic acid (sodium salt)
200
Presto FF-C18, 50 x 4.6 mm
MW: 4,000,000 - 6,000,000
0
0
5
mV
10 min
A: water / formic acid = 100 / 0.1
B: acetonitrile / formic acid = 100 / 0.1
0-30%B (0-10min)
0.4 mL/min (6MPa), 37 deg.C
ELSD (spray chamber 50 deg.C,
drift tube 100 deg.C)
2 uL (10ug, water)
MW: 1,500,000 - 2,500,000
200
Na+
Na+
Presto FF-C18, 150 x 4.6 mm
MW: 4,000,000 - 6,000,000
0
0
5
10
15 min
Imtakt USA ([email protected])
North America
A: water / formic acid = 100 / 0.1
B: acetonitrile / formic acid = 100 / 0.1
0-30%B (0-15min)
0.4 mL/min (16MPa), 37 deg.C
ELSD (spray chamber 50 deg.C,
drift tube 100 deg.C)
2 uL (10ug, water)
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI560E-JA08
No.TI567E
Gelatin
gelatin
(from bovine bone, ca.200 kDa)
mV
50
0
0
5
10
15 min
Presto FF-C18, 150 x 4.6mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
10-90 %B (0-15min), 0.4 mL/min (19MPa), 37 deg.C
ELSD (spray chamber 50deg.C, drift tube 100deg.C)
2 uL (10ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI567E-JA13
No.TI580E
Polyamino acids
NH2
H 3C
O
NH
CH
C
NH
n
poly-L-lysine
MW : 1,000-5,000
CH
O
C
n
poly-DL-alanine
MW : 1,000-5,000
1
mV
200
poly-L-lysine
MW : 1,000-5,000
2 uL(10ug)
2
100
poly-DL-alanine
MW : 1,000-5,000
3 uL(12ug)
0
0
10
20
30 min
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0-60 %B (0-20min)
0.2 mL/min (13MPa), 37 deg.C
ELSD (spray chamber 15 deg.C, drift tube 80 deg.C)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI580E-JB12
No.TI582E
Lectin (concanavalin A)
concanavalin A
(from Jack been, 25.5 kDa)
mV
200
A: water /formic acid = 100 /0.1
B: acetonitrile /formic acid = 100 /0.1
2 uL (5 ug)
100
concanavalin A
(from Jack been, 25.5 kDa)
A: water /trifluoroacetic acid
= 100 /0.1
B: acetonitrile /trifluoroacetic acid
= 100 /0.1
3 uL (7.5 ug)
0
0
5
10
15
20 min
Presto FF-C18, 150 x 4.6 mm
0-90 %B (0-15min)
0.4 mL/min (14 MPa), 50 deg.C
ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI582E-JB17
No.TI589E
PEGylated protein
mV
PEGylated protein, 60 kDa
(20 kDa protein + 40 kDa PEG)
200
30 x 4.6 mm
0-90 %B (0-2min)
0.5 mL/min (4 MPa), 1 uL
100
150 x 4.6 mm
0-90 %B (0-10min)
0.4 mL/min (14 MPa), 1.4 uL
0
0
5
10
15
20 min
Presto FF-C18
A: water / formic acid = 100 / 0.1
B: acetonitrile / formic acid = 100 / 0.1
50 deg.C, ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI589E-JB24
No.TI590E
PEGylated protein (2)
50 °C
PEGylated protein, 60 kDa
(20 kDa protein + 40 kDa PEG)
280nm
ELSD
0
10
20
30 min
Presto FF-C18, 150 x 4.6 mm
A: water /trifluoroacetic acid
= 100 / 0.1
B: acetonitrile /trifluoroacetic acid
= 100 / 0.1
20-70 %B (0-30min)
0.4 mL/min (15 MPa)
1 uL
37 °C
280nm
ELSD
0
10
20
Imtakt USA ([email protected])
North America
30 min
PEGylated protein, 60 kDa
(20 kDa protein + 40 kDa PEG)
Presto FF-C18, 150 x 4.6 mm
A: water /trifluoroacetic acid
= 100 / 0.1
B: acetonitrile /trifluoroacetic acid
= 100 / 0.1
20-70 %B (0-30min)
0.4 mL/min (19 MPa)
1 uL
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI590E-JB24
No.TI591E
mAU
PEGylated protein (3)
PEGylated protein, 60 kDa
(20 kDa protein + 40 kDa PEG)
40
50 °C
10 MPa
20
37 °C
13 MPa
0
0
10
20
30
40 min
Cadenza CW-C18, 250 x 3 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
50-57 %B (0-40min)
0.4 mL/min, 280 nm, 5 uL
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI591E-JD08
No.TI619E
mAU
Hemocyanin and hemoglobin
784 peaks
60
hemocyanin (from limulus)
35-50 %B (0-300min)
40
hemoglobin (from human)
37-39 %B (0-180min)
39-50 %B (180-300min)
20
683 peaks
0
0
100
200
300 min
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min (22 MPa), 50 deg.C, 220 nm
20 uL (100 ug, water or 0.02%TFA)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI619E-JG16
No.TI620E
mAU
Polyclonal IgG (from human serum)
30
Polyclonal IgG (human serum)
Oriental Yeast Co.,LTD
1205 peaks
20
10
0
0
100
200
300 min
15
10
Polyclonal IgG (human serum)
Sigma-Aldrich Corporation
1168 peaks
5
0
0
100
200
300 min
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
33-38 %B (0-300min)
0.4 mL/min (15MPa), 80 deg.C, 220nm
10 uL (30-100 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI620E-JG26
No.TI622E
mAU
Gragient slopes affecting mAb separation
20
30-45 %B (0-60min)
10
0
0
20
40
100
60
min
10-95 %B (0-20min)
0
0
10
20
min
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min (15MPa), 80 deg.C, 220nm
2 uL (anti-AFP, 2 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI622E-JG28
No.TI623E
mAU
Monoclonal and polyclonal antibodies
30
Monoclonal antibody
(anti-Derf2)
Monoclonal antibody
(anti-human LH)
20
Monoclonal antibody
(anti-AFP)
10
Polyclonal antibody
(human serum)
0
0
50
100 min
Presto FF-C18, 250 x 3 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
32-40 %B (0-120min)
0.2 mL/min (17MPa), 80 deg.C, 220nm
2-10 uL (2-10 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI623E-JH02
No.TI624E
mAU
The effect of column length on resolution for polyclonal IgG
Polyclonal IgG (human serum)
100
50 x 4.6 mm
187 peaks
31-36 %B (0-60min)
3.3 MPa
50
0
0
20
40
60 min
20
934 peaks
50 x 4.6 mm
31-36 %B (0-300min)
3.3 MPa
10
0
0
20
100
200
300 min
1205 peaks
250 x 4.6 mm
10
33-38 %B (0-300min)
15 MPa
0
0
100
200
300 min
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220nm, 10 uL (100 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI624E-JH20
No.TI626E
Gragient mixer volume affecting HSA separation
mAU
albumin (human serum, 66.4 kDa)
no mixer (T-union)
40
30
0.5mL mixer
20
1.7mL mixer
10
2.6mL mixer
0
0
20
40
60
80
100
120 min
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
30-40 %B (0-120 min)
0.4 mL/min (18 MPa), 80 deg.C, 220 nm, 1.2 uL (12 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI626E-KB02
No.TI627E
mAU
The effect of gragient slope and column length
for HSA separation
albumin (human serum, 66.4 kDa)
1000
50 x 4.6 mm
0-80 %B (0-10 min)
4 MPa
500
0
0
5
10
min
150
200
250 x 4.6 mm
0-80 %B (0-60 min)
16 MPa
100
0
0
20
40
60
min
20
250 x 4.6 mm
30-40 %B (0-120 min)
18 MPa
10
0
0
40
80
120 min
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 1.2 uL (12 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI62E-KB02
No.TI629E
The effect of gragient slope for angiotensin II separation
mAU
angiotensin II (human, 1046 Da)
2-52 %B (0-60 min)
50
0
0
20
40
60 min
80
120 min
10
2-12 %B (0-120 min)
0
0
40
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min (16 MPa), 80 deg.C, 220 nm, 1 uL (1 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI629E-KB02
No.TI630E
The effect of gragient slope for insulin separation
mAU
insulin
(bovine pancreas, 5733 Da)
100
2-52 %B (0-60 min)
0
0
20
40
60 min
80
120 min
10
21-31 %B (0-120 min)
0
0
40
Presto FF-C18, 250 x 4.6 mm
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min (16 MPa), 80 deg.C, 220 nm, 1.6 uL (1.6 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI630E-KB02
No.TI631E
mAU
The effect of gragient slope and column length
for BSA separation
albumin (bovine serum, 66.3 kDa)
1000
50 x 4.6 mm
0-80 %B (0-10 min)
4 MPa
500
0
0
5
10 min
150
250 x 4.6 mm
0-80 %B (0-60 min)
16 MPa
200
100
0
0
20
40
60 min
80
120 min
20
250 x 4.6 mm
30-40 %B (0-120 min)
18 MPa
10
0
0
40
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 0.6-1 uL (06-10 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI631E-KB02
No.TI632E
The effect of gragient slope and column length
for ovalbumin separation
mAU
albumin
(chicken egg, 45 kDa)
500
50 x 4.6 mm
0-80 %B (0-10 min)
4 MPa
250
0
0
150
5
10 min
250 x 4.6 mm
10-90 %B (0-60 min)
17 MPa
100
50
0
0
20
20
40
60 min
80
120 min
250 x 4.6 mm
40-50 %B (0-120 min)
17 MPa
10
0
0
40
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 1.6-4 uL (16-40 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI632E-KB02
No.TI633E
mAU
The effect of gragient slope and column length
for lectin separation
lectin (concanavalin A)
(jack been, 25.5 kDa)
400
50 x 4.6 mm
10-80 %B (0-10 min)
4 MPa
200
0
0
5
10 min
150
250 x 4.6 mm
10-80 %B (0-60 min)
16 MPa
100
50
0
0
20
40
60 min
80
120 min
250 x 4.6 mm
35-45 %B (0-120 min)
16 MPa
5
0
0
40
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 2-5 uL (2-5 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI633E-KB02
No.TI634E
mAU
The effect of gragient slope and column length
for thyroglobulin separation
thyroglobulin
(porcin thyroid, 670 kDa)
500
50 x 4.6 mm
5-95 %B (0-10 min)
4 MPa
250
0
0
10 min
5
250 x 4.6 mm
5-95 %B (0-60 min)
16 MPa
75
50
25
0
0
20
40
60 min
80
120 min
250 x 4.6 mm
30-60 %B (0-120 min)
18 MPa
20
10
0
0
40
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 10-20 uL (25-50 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI634E-KB02
No.TI635E
The effect of gragient slope and column length
for cytochrome c separation
mAU
cytochrome c
(horse heart, 12.4 kDa)
50 x 4.6 mm
0-80 %B (0-10 min)
4 MPa
500
0
0
5
10 min
150
250 x 4.6 mm
0-80 %B (0-60 min)
16 MPa
100
50
0
0
20
40
60 min
80
120 min
20
250 x 4.6 mm
21-31 %B (0-120 min)
17 MPa
10
0
0
40
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 0.6-3 uL (6-30 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI635E-KB02
No.TI636E
The effect of gragient slope and column length
for alpha-lactalbumin separation
mAU
alpha-lactalbumin
(bovine milk, 14.1 kDa)
500
50 x 4.6 mm
0-80 %B (0-10 min)
4 MPa
250
0
0
5
10
min
60
min
500
250 x 4.6 mm
0-90 %B (0-60 min)
16 MPa
250
0
0
20
40
20
250 x 4.6 mm
30-38 %B (0-120 min)
17 MPa
10
0
0
40
80
120 min
Presto FF-C18
A: water / trifluoroacetic acid = 100 / 0.1
B: acetonitrile / trifluoroacetic acid = 100 / 0.1
0.4 mL/min, 80 deg.C, 220 nm, 0.4-1.4 uL (4-14 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI636E-KB02
No.TI643E
mAU
Protein separation on non-porous ODS column (37°C, formic acid)
albumin (eggs, 45 kDa)
beta-glucosidase
(almond, 183 kDa)
thyroglobulin
(porcin thyroid, 670 kDa)
lectin (concanavalin A,
from Jack bean, 25.5 kDa)
alpha-chymotryosinogen A
(bovine pancreas, 25.7 kDa)
myoglobin
(equine skeletal muscle, -17 kDa)
alpha-lactalbumin
(bovine milk, 14.2 kDa)
100
papain (papaya latex, 21 kDa)
albumin (bovine, 66.3 kDa)
50
albumin (human serum, 66.4 kDa)
lactoferrin (bovine milk, 80 kDa)
lysozyme (chicken egg white, 14 kDa)
cytochrome c (horse heart, 12.4 kDa)
myosin (rabbit muscle)
alpha-casein (bovine milk, 23-26 kDa)
alpha-amylase (Bacillus sp., 50-55 kDa)
xanthine oxidase (buttermilk, 360 kDa)
0
0
10
20
30
min
Presto FF-C18, 75 x 4.6 mm
A: water / formic acid = 100 / 0.5, B: acetonitrile / formic acid = 100 / 0.5
10-50 %B (0-30min), 0.4 mL/min(10MPa)
37 deg.C, ELSD(spray chamber 35, drift tube 80 deg.C), 0.4-14 uL(0.8-10ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI643E-KB16
No.TI644E
mAU
Protein separation on non-porous ODS column (80°C, formic acid)
albumin (eggs, 45 kDa)
100
beta-glucosidase
(almond, 183 kDa)
thyroglobulin
(porcin thyroid, 670 kDa)
lectin (concanavalin A,
from Jack bean, 25.5 kDa)
alpha-chymotryosinogen A
(bovine pancreas, 25.7 kDa)
monoclonal antibody (anti-AFP)
myoglobin
(equine skeletal muscle, -17 kDa)
alpha-lactalbumin
(bovine milk, 14.2 kDa)
50
papain (papaya latex, 21 kDa)
albumin (bovine, 66.3 kDa)
albumin (human serum, 66.4 kDa)
lactoferrin (bovine milk, 80 kDa)
lysozyme (chicken egg white, 14 kDa)
cytochrome c (horse heart, 12.4 kDa)
0
0
10
20
30
min
Presto FF-C18, 75 x 4.6 mm
A: water / formic acid = 100 / 0.5, B: acetonitrile / formic acid = 100 / 0.5
10-50 %B (0-30min), 0.4 mL/min(5MPa)
80 deg.C, ELSD(spray chamber 35, drift tube 80 deg.C), 1-14 uL(2-14ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI644E-KB16
No.TI675E
Amyloid beta-protein
amyloid -protein
(Human, 4329 Da)
mV
20-40 %B (0-15min)
0.2 mL/min (6MPa)
60
20-40 %B (0-15min)
0.5 mL/min (4MPa)
5-25 %B (0-15min)
0.5 mL/min (3MPa)
0
0
5
10
15 min
100 x 4.6 mm
A: water / formic acid = 100 / 0.5
B: acetonitrile / formic acid = 100 / 0.5
37 deg.C
ELSD (spray chamber 35 deg.C, drift tube 70 deg.C)
1.4 uL (1.4 ug, DMSO)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI675E-KG08
No.TI684E
Cytochrome c ( 50°C, formic acid )
mV
cytochrome c
(horse heart, 12.4 kDa)
10
0
0
20
40
60
min
Presto FF-C18, 150 x 3 mm
A: water / formic acid = 100 / 0.5
B: acetonitrile / formic acid = 100 / 0.5
20-30 %B (0-60 min)
0.2 mL/min (17 MPa), 50 deg.C, ELSD (spray chamber 50 deg.C, drift tube 100 deg.C)
5 uL ( 50 ug)
Imtakt USA ([email protected])
North America
Imtakt Corp./JAPAN ([email protected])
Other Countries
TI684E-KH09