EQUIPO DE HPLC

Laboratorio de Control
de Calidad de la
Universidad Católica de
Santa María
PARTES DE UN HPLC BOMBA

HORNO DE COLUMNAS

AUTOSAMPLER

DETECTOR

RACK DE SOLVENTES
 TABLA1. AMINOÁCIDOS
Aminoácidos Pureza Gramos
DL. Acido Aspártico 98% 0.13835
DL. Ácido Glutámico 95% 0.14743
DL. Serina 99% 0.12233
Glicina USP Grade 0.75233
DL. Histidina 99% 0.15597
Clorhidrato de DL- Arginina CAS 0.21190
DL. Treonina 99% 0.11931
DL. Alanina 99% 0.89738
DL. Prolina 99% 0.11931
L. Tirosina 99% 0.89738
DL. Valina 99% 0.11686
DL. Metionina 99% 0.15023
DL. Isoleucina 99% 0.13201
L. Leucina USP- Grade 0.13238
DL. Fenilalanina 99% 0.16971
L. Triptofano 99% 0.20922
 METODOLOGÍA
SOLUCIÓN ETANOL,
AGUA, LIOFILIZACION
TRIMETILAMINA Y COMPLETA
PREPARACIÓN DE FENILTIOCIANATO
SOLUCIONES STOCK 7:1:1:1.

SOLUCION: ETANOL, DILUCIONES DE

AGUA Y 0.25, 0.50, 0.75 Y 1
[0.1M] TRIMETILAMINA, EN uM/ml
BUFFER FOSFATO LAS PROPORCIONES
DE 2:2:1,

LECTURA EN EL
HPLC
FORMACIÓN DE AA. DERIVATIZACION
1uM/ml DE AMINOACIDOS
 mAU

0.423
0.553

0
20
40
60
80
100
120
140
160
180

-20
0.787 1055167084
6262
437
1.18717248

0
1.600
1.843
1.977 2888
2.103
2.180
2.260 7515
2928
10006
1645
2.343 2588
2039

Area
2.530 24755

Name
2.847
3.120
3.237 1653
11215
6334

2
3.503
3.753
3.827
3.897 1451
1682
2185
3.987 3885
4.273
4.357
4.500 3946
2052
4.593 2980

Retention Time
4.667
4.737 1531
24142
1757
1497
5.027

4
183511

Spectrum Max Plot

6.513 30341
6.650 225789
402804
7.247

6
7.527

26585
18547
8.647
8.777 61340
24973
9.120
9.247 29783
126562

8
9.377
9.723 7895067
1085763 10.497
11.097

10
149349
12.363
12.610
12.750 29939
13.037 54810
13.100
13.243
13.310 22070
34006
19747
13.537 22855
45622
111293
Primera Corrida Continuación 004 04/10/2017 09:36:41 p.m.

12
13.927
14.067
14.147 21738
14.347 52883
32469
14.517
14.583
14.657
14.760 26729
14311
33735
14.913 14518
52047
147429
PICO DEL AA.

15.500
15.580
15.633 22416
15.783 25065

14
15.873
16.020 30762
47220
34509
16.133 29656
964646
16.843
103017
265496

16
17.890
17.970
18.693
18.797 26797
26644
18.913 65485
208014
19.327 102209
19.643
19.743 35469

18
20.063
20.133
20.213 21638
29166
20.340
20.430
20.497 17232
41957
28042
29184

726912

20
21.803

1111598
23.567
23.653 550444

22
23.943 51227
318370
371166
24.510
24.713 225032
25.060 347486
25.363 339291
25.657
13886022 675474

24
25.863
358475
26.353
26.797

26
2825664
28.617

Minutes
568224
29.650

28
372768
809110
30.290
30.503
759633
31.250

30
3175815
32.740 317844
800102
32.877 428546
1119942
33.113
33.230 910150
33.400 622916
33.533 2538347
33.697 258942

32
1381129
AMINOÁCIDO GLICINA

33.950 1176720
79375834.177
34.380
34.487 1869575 3674909
34.627 4852939
1948531
35.100 34.910
4569560 35.393
35.693

34
3860143
36.070
10175611
36.530
37.347

36
38
40
PICOS DE LOS SOLVENTES.

42.197
42.410
42.487
42.640 5060
42.793
42.867 1278
43.053
43.173 695
43.273
43.340
43.433
43.490 564
1352
43.727 697
448
43.830
42

44.073
44.157 512
296
275
44.267 557
589
44.717 924
170
44.940
45.033 234
611
249
45.090
45.363
45.447
45.530 1074
45.610
45.703 1034
45.807 51
495
44

46.020 646
46.320 406
322
292
46.493
46.607 662
217
903
2117
47.170 662
47.477
47.547 593
3096
47.640
46

47.893
48.057 1275
48.547 365
151
48.597
48.700 363
48.830
48.990 112
49.160
49.220
49.323
49.573 158
239
795
49.680 68
628
48

49.813 870
639
2928
811
1853
50.283
50.333
50.497 992
6923
2745
50.983
51.170
51.293
51.340 155
4070
51.687 2152
50

52.040
52.147 253
238
52.253
52.467
52.577
52.650
52.913 82
760
552
53.137
53.240 273
526
758
1211
923
53.617
52

53.870 921
614
1513
604
0
20
40
60
80

-20
100
120
140
160
180

mAU