Data File C:\CHEM32\...\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\007-0702.

D
Sample Name: MTRA GRUPO 3-2

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PROYECTO
FARMACOCINÉTICA POBLACIONAL DE PIPERACILINA EN UNIDADES DE CUIDADO INTENSIVO

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REPORTE DE MÉTODO
LABORATORIO DE ANÁLISIS INSTRUMENTAL FARMACÉUTICO
VALIDACIÓN DE METODOLOGÍA BIOANALÍTICA

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=====================================================================
Acq. Operator : ASANCHEZ_EJNINOL Seq. Line : 7
Acq. Instrument : HPLC Anàlisis Instrumental Location : Vial 7
Injection Date : 2/6/2019 9:47:34 PM Inj : 2
Inj Volume : 10 µl
Sequence File : C:\Chem32\1\DATA\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\CC
CIPRO 2019.S
Method : C:\Chem32\1\DATA\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\
CIPROFLOX.M
Last changed : 2/6/2019 5:05:41 PM by ASANCHEZ_EJNINOL
Method Info : Condiciones cromatográficas VALIDACIÓN DE MÉTODOS
Columna LiChrospher

=====================================================================
Module Type Firmware rev. Serial number
--------------------------------------|------|---------------|---------------
1100/1200 Column Thermostat G1316A A.06.04 [001] DE63061888
1100/1200 Quaternary Pump G1311A A.06.04 [001] DE62959955
1100/1200 Diode Array Detector G1315B A.06.04 [001] DE63057980
1200 Autosampler SL G1329B A.06.32 [004] DEABE00746

Software Revision: Rev. B.03.02 [341] Copyright © Agilent Technologies

=====================================================================

Column Description : ODS-2 Hypersil

Product# : 31605-254630 Batch#: 15925
Serial# : 10771820
Diameter : 4.6 mm Length : 250.0 mm
Particle size : 5.0 µm Void volume : 2.50 ml
Maximum Pressure : 400 bar Maximum pH : 9
Maximum Temperature: 60 °C

HPLC Anàlisis Instrumental 2/6/2019 10:03:01 PM ASANCHEZ_EJNINOL Page 1 of 5

Data File C:\CHEM32\...\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\007-0702.D
Sample Name: MTRA GRUPO 3-2

Comment : pHmin 2.0

=====================================================================

Instrument Conditions : At Start At Stop

Column Temp. (left) : 30.0 30.0 °C
Column Temp. (right) : 30.0 30.0 °C
Pressure : 169.7 171.0 bar
Flow : 1.500 1.500 ml/min

Detector Lamp Burn Times: Current On-Time Accumulated On-Time

DAD 1, UV Lamp : 6.64 2291.1 h
DAD 1, Visible Lamp : OFF 4812.0 h

Solvent Description :
PMP1 , Solvent A : H2O
PMP1 , Solvent B : ACN
PMP1 , Solvent C : FM
PMP1 , Solvent D : MeOH
=====================================================================
Run Logbook
=====================================================================
Method Method started: line# 7 vial# 7 inj# 2 21:46:45 02/06/19
Method Instrument running sample Vial 7 21:46:45 02/06/19
PUMP 1 Pressure = 169.7 bar 21:47:46 02/06/19
THM 1 Column temperature = 30.0 °C 21:47:46 02/06/19
PUMP 1 Pressure = 171.0 bar 22:02:46 02/06/19
THM 1 Column temperature = 30.0 °C 22:02:46 02/06/19
Method Saving Method CIPROFLOX.M 22:02:50 02/06/19
Method Saving Method RUN.M 22:02:53 02/06/19
CP Macro Analyzing rawdata 007-0702.D 22:02:54 02/06/19
Method Instrument run completed 22:03:00 02/06/19
=====================================================================

HPLC Anàlisis Instrumental 2/6/2019 10:03:01 PM ASANCHEZ_EJNINOL Page 2 of 5

Data File C:\CHEM32\...\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\007-0702.D
Sample Name: MTRA GRUPO 3-2

DAD1 A, Sig=278,30 Ref=320,50 (007-0702.D)

8.879
mAU

120

100

80

60

40

20
2.874

0 2 4 6 8 10 12 14 min
PMP1 , Solvent B
%

0.08

0.06

0.04

0.02

-0.02

-0.04

-0.06

-0.08

-0.1
0 2 4 6 8 10 12 14 min

HPLC Anàlisis Instrumental 2/6/2019 10:03:01 PM ASANCHEZ_EJNINOL Page 3 of 5

Data File C:\CHEM32\...\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\007-0702.D
Sample Name: MTRA GRUPO 3-2

=====================================================================
Internal Standard Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Use Multiplier & Dilution Factor with ISTDs

Signal 1: DAD1 A, Sig=278,30 Ref=320,50

=====================================================================
=====================================================================
Area Percent Report
=====================================================================

Sorted By : Signal
Multiplier : 1.0000
Dilution : 1.0000
Use Multiplier & Dilution Factor with ISTDs

Signal 1: DAD1 A, Sig=278,30 Ref=320,50

Peak RetTime Type Width Area Height Area

# [min] [min] [mAU*s] [mAU] %
----|-------|----|-------|----------|----------|--------|
1 2.874 BB 0.0707 27.32484 5.94057 0.7420
2 8.879 BB 0.4152 3655.25635 124.27095 99.2580

Totals : 3682.58119 130.21152

=====================================================================
=====================================================================
Calibration Curves
=====================================================================
=====================================================================

HPLC Anàlisis Instrumental 2/6/2019 10:03:01 PM ASANCHEZ_EJNINOL Page 4 of 5

Data File C:\CHEM32\...\2019_FINAL_DEFINITIVO\CC CIPRO 2019 2019-02-06 17-08-14\007-0702.D
Sample Name: MTRA GRUPO 3-2

Signal DAD1 A, Sig=278,30 Ref=320,50 (007-0702.D)

mAU

100
80
60
40
20
0
0 2 4 6 8 10 12 14 min
__________________________________________________________________________________________________
Peak :1 at 2.874 min Name : ?

| |
| ' ' ' ' ' |

Calculated

----------------------------++++++
220 240 260 280 300 320 340 360 380 nm 2.8 2.9 3 min

-> The purity factor exceeds the calculated threshold limit. <-

Purity factor : 999.506 (6 of 34 spectra exceed the calculated threshold limit.)

Threshold : 999.693 (Calculated with 6 of 34 spectra)
Reference : Peak start and end spectra (integrated) (2.789 / 3.016)
Spectra : 5 (Selection automatic, 5)
Spectra : Smooth 7, Splined 5
Noise Threshol: 0.040 (12 spectra, St.Dev 0.0188 + 3 * 0.0072)
__________________________________________________________________________________________________
Peak :2 at 8.879 min Name : ?

| |
| ' ' ' ' ' |

Calculated

+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
220 240 260 280 300 320 340 360 380 nm 8.5 9 9.5 10 10.5 min

-> The purity factor is within the calculated threshold limit. <-

Purity factor : 999.924 (487 of 487 spectra are within the calculated threshold limit.)
Threshold : 999.892 (Calculated with 487 of 487 spectra)
Reference : Peak start and end spectra (integrated) (8.459 / 10.519)
Spectra : 5 (Selection automatic, 5)
Spectra : Smooth 7, Splined 5
Noise Threshol: 0.040 (12 spectra, St.Dev 0.0188 + 3 * 0.0072)
__________________________________________________________________________________________________
*** End of Report ***

HPLC Anàlisis Instrumental 2/6/2019 10:03:01 PM ASANCHEZ_EJNINOL Page 5 of 5