OpenLAB CDS
REVOLUTIONIZE
YOUROpenLAB
DESKTOPCDS ChemStation
Integration
of Agilent
SEQUENCING WORKFLOW
Edition
with OpenLAB Data Store
HALOPLEX TARGET
ENRICHMENT with
SYSTEM
Compliance
21 CFR Part 11
What a Difference a Day Makes
Complete Target Enrichment in Less Than 6 hours
Technical Note
Highlights
Summary
• Expanded capture size – enrich thousands
of targets, all in a single tube
• Scalable protocols – 96-samples ready for
sequencing in less than a day, with Agilent
automation solutions
The Agilent HaloPlex Target Enrichment System enables fast, simple, and efficient
analysis of genomic regions of interest for a large number of samples, covering
thousands of exons per sample. It uses single-tube target amplification and removes
Introduction
the need for library preparation to reduce total sample processing time and cost
without the need for dedicated instrumentation. HaloPlex is compatible with
Part
11 in Title
21 ofsequencing
the Code of Federal
Regulations includes
the USItFederal
guidelines
different
desktop
and high-throughput
platforms.
also expands
your
for
storingwith
and protecting
electronic
andand
applying
electronic signatures.
options,
the availability
of 96records
indexes
an automation
protocol, to deliver the
flexibility you need to efficiently run your experiments. Finally, you can access an
The intent of these guidelines is to ensure that applicable electronic records are reliable,
assay with premium performance, high specificity, uniformity, and variant detection.
authentic and maintained with high integrity. This technical note describes features and
• Performance and reliability – greater
specificity, uniformity, and variant calling
combination with Agilent OpenLAB Data Store for data management, which enable them
• Intuitive Design Wizard – create panels
in less than 10 minutes
• Compatibility with desktop sequencers
and high-throughput NGS platforms
• Immediate implementation – no dedicated
capital equipment required
PREMIUM PERFORMANCE
functionality
of Agilent OpenLAB CDS ChemStation Edition, revision C.01.04 or higher, in
Design Size Range
Up to 500Kb (>2,500 exons); 1.5Mb coming soon
Specificity (% bases on-target)
> 99 %*
Uniformity (coverage at 10 % of mean)
> 90 %*
% Bases covered at 1X
> 98 %*
% Bases covered at 10X
> 97 %*
to meet the guidelines of 21 CFR Part 11.
This document examines each section of 21 CFR Part 11 and provides a recommended
remediation
approach using the integration of OpenLAB
CDS ChemStation Edition rev.
% Bases covered at 20X
> 96 %*
C.01.04
with
OpenLAB
Datadesigns,
Store.
*Performance
dataAgilent
was collected
for 12 different
with sizes ranging from 4.4kb to 400kb.
100%
Agilent
OpenLAB CDS ChemStation Edition rev. C.01.04 provides a tight and seamless
80%
integration into Agilent OpenLAB Data Store. This integrated solution provides the necessary
60%
controls for managing system access, user management, data transfer handling and audit
40%
trail
functionality, while Agilent OpenLAB Data Store ensures secure record keeping.
20%
0%
Covered
by Design
4.4 Kb
% Bases
on Target
21 Kb
60 Kb
% Bases
Covered 1x
120 Kb
% Bases
Covered 20x
280 Kb
396 Kb
% Bases
Covered 20x
% Bases
% Bases
Covered at
Covered at
10% of Mean 20% of Mean
Identification of Unknown Organic Pollutants
A key strength of the method described here is its ability to simultaneously identify
Phenazone: +ESI EIC(189.1022)
x104 SNR (4.018min) = 68.1
4.018
3
21
CFR Part
11 sections
addressed
byinthe
integration
of Agilent OpenLAB
CDS ChemStation
unexpected
organic
pollutants. As
demonstrated
Figure
3, an unexpected
2.5
contaminant,
metoprolol,
was
identifi
ed
by
accurate
mass
measurement
and
isotope
Edition (revision C.01.04 or higher) with Agilent OpenLAB Data Store (revision
A.01.01 or higher)
2
pattern matching. The identity of metoprolol is further confirmed by matching its
MS/MS spectrum to the Broecker, Herre & Pragst Forensics/Toxicology Personal
1.5
Compound Database and Library (Figure 4).
The integration of OpenLAB1 CDS ChemStation Edition with OpenLAB
Possible scenarios with
Electronic
record only
Electronic
signature based
3
3
Data Store enables the operation of OpenLAB CDS ChemStation
2
0.5
ChemStation
+ESI Productoperated
Ion (2.752, in
2.758 min, 2 Scans)
Frag=225.0V CID@20.2
(268.1908[z=1]
-> **)
x10
(without
upon User
ID
Edition268.1909
in full support of all compliance requirements mandated by
72.0805
a1closed system
signature)
and Password
0.75
21 CFR Part 11 for a closed0 system. In particular it ensures:
2.5
3
3.5
4
4.5
5
5.5
0.5
148.0866
11.1, 11.2, 11.3
Counts vs. Acquisition time (min)
191.1078
0.25
(Scope, implementation, definition)
• Accurate and complete copies of records
0
Figure 2. Extracted ion chromatogram of
2
Ion (2.752, 2.758 min, 2 Scans) Frag=225.0V CID@20.2 (268.1908[z=1] -> **)
x10
11.10+ESI Product
phenazone
10 ng/L level, with the
268.1909of all relevant
72.0805
• Versioning
recordsatforthe
traceability
1
(Controls
for closed systems)
structure of the compound (inset).
148.0866
0.5
191.1078
0
• P reservation of records between their creation and their automatic
11.30
-0.5
N/A
N/A
(Controls
for open systems)
-1
transfer to OpenLAB Data Store immediately after acquisition,
3
3
x102 Metoprolol C15H25NO3 + Product Ion Frag=150.0V CID@20.0 ForensicsTox_AM_PCDL.cdb reprocessing or interactive4 modifications
268.1907
x10 Metoprolol: +ESI Scan
11.50
1
N/A
268.1907
1.2
74.0600
(Signature manifestations)
116.1070
0.75
98.0964
• Controlled copies of the1.1
data (M+H)+
0.5
133.0648 159.0804
191.1067
176.1070
1
0.25
148.0757
226.1438 250.1802
11.70
91.0542
N/A
0
0.9
(Signature
record linking)
250andatory
Data Store before allowing
60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240• M
260 270 login to OpenLAB
0.8
Counts vs. Mass-to-Charge (m/z)
appropriate
access
to
the
ChemStation
0.7
11.100
N/A
0.6
(e-Sig general requirements)
Figure 4. Identification of metoprolol can be further confirmed by MS/MS and retention time
0.5
•
R
ecords
of
changes
captured
in user-independent time-stamped
matching to a reference compound (top: sample spectrum, middle: mirror spectrum, bottom:
0.4
11.200 (a)
269.1940
audit
trails
N/A
library
spectrum).
0.3
(e-Sig not biometric)
(M+H)+
0.2
270.1965
0.1
These settings can be configured
during or after installation(M+H)+
to meet
11.200 (b)
N/A
N/A
0
(e-Sig
biometric)
specific standard operating procedures
and 269
security
guidelines.
Broad Applicability
268 268.5
269.5
270 270.5
3
3
3
3
This includes customizable ChemStation
security
roles and privileges
Counts
vs. Mass-to-Charge
(m/z)
11.300
(a),Q-TOF
(b), (d) with Jet Stream and iFunnel technology delivers the utmost
The
6550
N/A
3
(Controls for ID codes / passwords)
that provide restricted access
to
ChemStation
functionality
to
Figure 3. EIC of an unexpected contaminant,
sensitivity for a broad range of target compounds (Table 1), enabling ultra-trace
metoprolol,
a signal-to-noise
ratio of 46:1.
individual users or groups.
Changeswith
to the
security configuration
11.300 (c), (e)
compound
identification and quantitation.
Red squares represent the theoretical isotope
N/A
N/A
can be made only by a dedicated system administrator. Details and
(Token cards & other ID devices)
intensity and position.
recommendations for configuration of the system are outlined in
Table 1:
Table 1. LOD and LOQ values (ng/L) for a broad range of target compounds.
the manual “OpenLAB CDS ChemStation Edition with Central Data
Applicable sections of 21 CFR Part 11 for OpenLAB CDS ChemStation Edition/OpenLAB
Data
Store
operated
in
a
closed
system
(
3
=
applicable,
N/A
=
not
applicable
Storage”,
Part Number M8301-90081, which is available on the
Compound Name
LOD LOQ
Compound Name
LOD LOQ
OpenLAB
CDS ChemStation Edition installation media.
Atenolol
0.84
3.59
Sulfamethoxazole
2.97
10.84
Sotalol
0.44
2.01
Carbamazepine 10,11 epoxide
0.82
3.52
Metronidazole
0.73
3.19
Phenacetin
0.81
3.50
Trimethoprim
3.02
11.02
Carbamazepine
0.53
2.39
Sulfamerazine
2.55
9.39
Betaxolol
1.06
4.40
Sulfadimidine
2.59
9.53
Phenazone
0.47
2.14
n,n Diethyl-3-methylbenzamide
(DEET)
0.95
4.02
Pentoxifylline
1.07
4.47
Bezafibrate
1.12
4.62
Metoprolol
0.56
2.51
Crotamitron
0.49
2.22
Furazolidone
2.27
8.47
Diazepam
0.43
1.97
Dapson
3.85
14.70
Fenofibrate
3.35
12.31
The integration of OpenLAB CDS ChemStation Edition rev. C.01.04 with OpenLAB Data Store was not specifically designed for operation in an open system.
NOTE:
In this technical note the2.65
term “ChemStation”
(in the context of integration
Ronidazole
9.73
Bisoprolol
1.38 with
5.52OpenLAB Data Store) always refers to OpenLAB CDS
ChemStation
feature set for the integration
OpenLAB Data Store enabled.
Sulfadiazine Edition rev. C.01.04
2.69or higher
9.87 with the
Propranolol
1.13 with
4.67
*Calculated based on the DIN 32645 using a program courtesy of Dr. Joachim Kleiner. (www.kleiner-j.de)
www.agilent.com/lifesciences/QTOF
For Research Use Only. Not for use in diagnostic procedures.
This information is subject to change without notice.
© Agilent Technologies, Inc. 2012
Published in the USA, April 5, 2012
5990-9862EN
271
Identification of Unknown Organic Pollutants
Phenazone: +ESI EIC(189.1022)
11.10 Controls for Closed Systems
x104 SNR (4.018min) = 68.1
A key strength of the method described here is its ability to simultaneously identify
4.018
3
Section
Question
OpenLAB CDS ChemStation Edition / OpenLAB Data Store
Integration
unexpected organic pollutants. As demonstrated in Figure 3, an unexpected
2.5
Yes. Agilent Technologies has extensively validated the performance of ChemStation
/ OpenLAB Data Store with tests written to
contaminant,
was identifi
massaccuracy,
measurement
and isotope
Hasmetoprolol,
the system been validated
in order ed by accurate
specifically evaluate
reliability and consistent
performance. OpenLAB Data Store maintains revisions of files as they are
2
to
ensure
accuracy,
reliability,
consistent
or altered.
The solution
any and all
pattern
The identity of metoprololmodified
is further
confi
rmedalso
byrecords
matching
itschanges made to the system through computer generated audit trails.
11.10(a) matching.
intended performance, and the ability to
Agilent
Technologies
recommends
making
use
of
Installation
discern invalid
altered
records? Herre & Pragst Forensics/Toxicology Personal Qualification and
MS/MS spectrum
to orthe
Broecker,
1.5Operation Qualification (IQ/OQ) service for ChemStation
/ OpenLAB Data Store to validate the on-site system per compliance requirements.
Compound Database and Library (Figure 4).
1
Yes. OpenLAB Data Store centrally stores all data types, from raw machine data to printable reports. All files are unaltered and stored
in the original format.
Is the Ion
system
capable
of generating
accurate
0.5
(2.752,
2.758
min, 2 Scans)
Frag=225.0V CID@20.2 (268.1908[z=1] -> **)
x102 +ESI Product
and complete copies of all required records
Raw data, metadata and result data generated by268.1909
ChemStation are stored and managed in OpenLAB Data Store. The result set
72.0805
1
in both human readable and electronic form
11.10(b)
0 PC as a copy of the original data for review. The
that holds all this information can be loaded at any time to the hard disk of a client
0.75
suitable for inspection, review and copying
ChemStation is required to read the electronic format. “Printed” reports, representing the
2.5human-readable
3
3.5 form4of electronic
4.5 records,
5
5.5
0.5
by the FDA?
148.0866 can be stored as PDF files which can be made available for review without the source application
installed
on the client time
machine.
Counts
vs. Acquisition
(min)
191.1078
0.25
These reports can include all data and audit trails. A viewer is available to view these PDF reports without the original application.
0
Figure 2. Extracted ion chromatogram of
CID@20.2
(268.1908[z=1]
->data
**) generated by the ChemStation are stored and managed in OpenLAB Data Store. Data stored
Yes. Raw
data, meta
data and result
x102 +ESI Product Ion (2.752, 2.758 min, 2 Scans) Frag=225.0V
at the
10tong/L
level,storage
withlocation
the of
268.1909
72.0805
in OpenLAB Data Store resides in a protected storage
location. The individualphenazone
users do not have
access
the physical
1
structure of the compound (inset).
148.0866 the files. 191.1078
0.5
Are the records protected for accurate
0
ChemStation can be configured to store all raw data, metadata and result data automatically in OpenLAB Data Store immediately after
-0.5
and convenient retrieval throughout the
11.10(c)
acquisition and after each interactive review or automated reprocessing.
-1
record retention period?
2
The functionality
is complemented by additional procedural controls that should be defined and implemented by the system administrator
ForensicsTox_AM_PCDL.cdb
x10 Metoprolol C15H25NO3 + Product Ion Frag=150.0V CID@20.0
Metoprolol:
+ESI
based on company-wide security policies. These policies
should manage practices
as access to
clientScan
computers and password
268.1907
x104 such
1
renewal
frequency.
268.1907
1.2
74.0600
116.1070
0.75
(M+H)+
98.0964
1.1
0.5
133.0648 159.0804
Yes. Each 191.1067
user is identified by a unique user ID and password combination. Access to ChemStation / OpenLAB Data Store requires
176.1070
1
0.25
148.0757
226.1438
91.0542
250.1802
the entry of both identification components:
user ID and password. In ChemStation / OpenLAB Data Store, all file and software
0
functionality access is controlled by privileges and roles assigned to individual0.9
users or groups of users. The system administrator
60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270
0.8
determines levels of access.
Counts vs. Mass-to-Charge (m/z)
0.7
Both Data Store- and ChemStation privileges are assigned in the OpenLAB Shared Services. A set of more than 40 ChemStation
Is system access limited to authorized
0.6
11.10(d)
privileges
is
available
in
OpenLAB
Shared
Services
to
allow
limited
system
access
to authorized individuals and control the access level
individuals?
Figure 4. Identifi
cation of metoprolol can be further confirmed by MS/MS and retention time
0.5 user roles. The ChemStation access privileges offer
of different user roles. These privileges can be combined when defining individual
matching to a reference compound (top: sample spectrum,
middle:
mirror
spectrum,
bottom:
0.4pre-defined user roles for ChemStation access levels.
granular access restriction to ChemStation functionality. The system offers four
269.1940
library spectrum).
The management of users, roles and privileges is handled in OpenLAB Shared0.3
Services, where user management
(M+H)+can be integrated
with Windows user management. Depending upon access restrictions, menu0.2
items, graphical elements or views in the ChemStation
270.1965
can be enabled or disabled.
(M+H)+
0.1
0
Yes. All actions related to creating, modifying or deleting electronic records are recorded in a secure, computer-generated, timeBroad Applicability
268 268.5 269
269.5 270 270.5
stamped audit trail. The audit trail lists all modifications, date and time of the change, the user ID and reason for the change if
Counts
vs.
Mass-to-Charge
(m/z)
Entries in the audit trails cannot be switched off, altered or deleted by the user. Even the removal of records from OpenLAB
The 6550 Q-TOF with Jet Stream and iFunnelapplicable.
technology
delivers
thenotutmost
Data
Store by an authorized
user does
affect existing entries in the audit trail.
Figure 3. EIC of an unexpected contaminant,
sensitivity for a broad range of target compounds
(Table/ OpenLAB
1), enabling
The ChemStation
Data Storeultra-trace
integration ensures that all meta data
is stored along
with
data and result data.
metoprolol,
with
a raw
signal-to-noise
ratioThe
ofresult
46:1.
or single runrepresent
is initiated, the
and theisotope
sequence
compound identification and quantitation. set and single run of ChemStation maintain full data integrity. When a sequence
Red squares
themethods
theoretical
template associated with the run(s) are copied to the result set or single run. The stored information includes all audit trails related to
and position.
the result data and the method(s) used for data generation. All changes to intensity
a running sequence
or single run are captured as well.
/ OpenLAB Data Store integration provides three types of audit trails:
Table 1. LOD and LOQ values (ng/L) for a broad rangeTheofChemStation
target compounds.
Compound Name
Atenolol
11.10(e)
Sotalol
LOD
LOQ
Is there a secure, computer-generated
0.84records
3.59
audit trail that independently
the date
and time of operator entries
and
actions
0.44
2.01 that
create, modify, or delete electronic records?
1. O
penLAB Data Store Audit Trail: The OpenLAB Data Store audit trail records who has accessed the system and what operations he/
Compound
Nameduring a given period
LOD
she has performed
of time. LOQ
The recorded activities include items such as data storage, versioning, and electronic
signatures. Removing records from the database does not affect existing entries in the audit trail. Logon to the ChemStation is
Sulfamethoxazole
2.97
10.84
similarly documented.
Carbamazepine
10,11
epoxide
0.82consists
3.52
2. Method Audit Trail:
The method
audit trail
of two components: a) the method management concept which saves used
methods along with the data files, introduced
ChemStation Revision B.02.01, and b) the method audit trail.
Phenacetin
0.81 with
3.50
Metronidazole
0.73
3.19
Ronidazole
2.65
9.73
The method audit trail tracks all changes
to the data
acquisition and data analysis parameters. The system enforces user-added
Bisoprolol
1.38
5.52
Sulfadiazine
2.69
9.87
tracked with the user ID, date and time1.13
of the method
Propranolol
4.67modification, and user comments. The audit trail documents the changes from
Trimethoprim
3.02
11.02
Carbamazepine
Sulfamerazine
2.55
9.39
Sulfadimidine
2.59
9.53
Phenazone
0.47
2.14
Pentoxifylline
1.07
4.47
Metoprolol
0.56
2.51
Furazolidone
2.27
8.47
When records are changed, is previously
11.10(e)
recorded information left3.85
unchanged?
Dapson
14.70
comments for any changes to the data acquisition parameters. Any changes to the data analysis parameters are automatically
one “save” to the next.
0.53
2.39
3. R
esults Audit Trail: A result-specific audit trail contains a log of changes that occurred when a user worked on one revision of a run.
Betaxolol
1.06are actively
4.40 saved to the data file and the logbook tracks all result changes from one
Manual integration activities and changes
“save” to the next. Manual integration is automatically tracked with the user ID, date and time, and the steps that were performed
n,n Diethyl-3-methylbenzamide
to modify the integration.
0.95
4.02
(DEET)
In addition, the OpenLAB Data Store audit trail also keeps a record of all changes to the OpenLAB Data Store system, including
Bezafi
brateremoval or changes of folders.
1.12
additions,
4.62
www.agilent.com/lifesciences/QTOF
Crotamitron
0.49
2.22
Yes. All entries in the ChemStation / Data Store audit trails are non-editable and non-deletable. Even the removal of records from
OpenLAB Data Store by an authorized user
does not1.97
affect existing entries
in the auditUse
trail.Only. Not for use in diagnostic procedures.
For Research
Diazepam
0.43
This information is subject to change without notice.
Strict revision
by the ChemStation
is achieved by forcing automatic storage of the result set or single
Fenofi
brate control of the data generated
3.35
12.31
runs in OpenLAB Data Store after acquisition, automatic reprocessing or any other interactive change. Records are securely stored
© Agilent Technologies, Inc. 2012
in a protected transfer queue that cannot be accessed by unauthorized users. This prevents any alteration of records between their
Published in the USA, April 5, 2012
*Calculated based on the DIN 32645 using a program courtesy of Dr. Joachim Kleiner. (www.kleiner-j.de)
creation or modification in ChemStation and storage in OpenLAB Data Store as may occur, for example during a network outage.
5990-9862EN
271
Yes. All OpenLAB Data Store audit trail information is stored in the OpenLAB Data Phenazone:
Store repository
as part
of a file´s meta data and
Identification of Unknown Organic Pollutants
+ESI
EIC(189.1022)
kept throughout the electronic records retention period. The OpenLAB Data Store
trails are unbreakably linked to the record.
4 audit
SNR (4.018min) = 68.1
Are electronic audit trails saved at least as
System-related activities such as logon events are unbreakably linked to thex10
system.
A11.10(e)
key strength
oftheir
the
method
described
4.018
long as
subject
electronic
records and here is its ability to simultaneously identify
3
for agency review and reproduction? The results audit trail is part of the run logbook and is stored with each data file. The method audit trail is part of the method and
unexpected available
organic
pollutants. As demonstrated
in Figure
3, an
unexpected
is stored with
each method
file. Both,
data file and method are stored in OpenLAB
Data
Store.
This
ensures
an
unbreakable link
2.5
between record
and related
audit trail.
contaminant, metoprolol, was identified by accurate
mass
measurement
and isotope
pattern matching. The identity of metoprololYes.
isInfurther
confiand
rmed
by Data
matching
its when a sequencing2 of events is required, system checks enforce it.
all ChemStation
OpenLAB
Store functions,
A few examples are:
MS/MS spectrum to the Broecker, Herre & Pragst Forensics/Toxicology Personal
1.5
approved methods are to be used in QA/QC, this can be achieved by restricting user access to the approved methods stored
Compound Database and Library (Figure 4). • inIf only
OpenLAB Data Store.
1
Are operational system checks used to
enforce permitted sequencing of steps
• In ChemStation, Agilent uses a clear prerun/run/postrun structure in its ‘Run Control’ functionality to ensure proper sequencing
0.5
Ion (2.752, 2.758 min, 2 Scans) Frag=225.0V
CID@20.2 (268.1908[z=1] -> **)
and events?
x102 +ESI Product
of steps.
268.1909
72.0805
1
The ChemStation and Data Store user privileges are entirely managed by a system
0 administrator and restrict the user to an
0.75
automatic transfer of the records from ChemStation to OpenLAB Data Store. This ensures
always5
2.5that ChemStation
3
3.5 records
4 are4.5
5.5
0.5
148.0866 stored in OpenLAB Data Store.
Counts vs. Acquisition time (min)
191.1078
0.25
0
Are authority checks in place to ensure
Yes. Users cannot gain access to OpenLAB Data Store without a valid userFigure
ID, and password.
Only a ion
successful
logon to the system
2. Extracted
chromatogram
of
that only
individuals
use Frag=225.0V CID@20.2 (268.1908[z=1] -> **)
Ionauthorized
(2.752, 2.758
min, 2can
Scans)
x102 +ESI Product
offers access to files and general software functionality,
chromatographic phenazone
software functions
and 10
approval
Thisthe
includes
at the
ng/Lfunctionality.
level, with
268.1909
72.0805
the
system,
electronically
sign
a
record,
1
file signing, value input, record alteration, and other practices. The user must authenticate with a valid user ID and password. This
11.10(g)
structure
of
the
compound
(inset).
access
the
operation
or
computer
system
148.0866 applies at 191.1078
0.5
program initiation and after every inactivity timeout on the computer program. User-access to specific functionality in the
input or output device, alter a record, or
0
software is further restricted by the privileges assigned to the individual user. These privileges can be combined into roles if necessary.
perform
the
current
operation?
-0.5
-1
Yes. For ChemStation, instrument serial numbers are transferred electronically. This is completed automatically from the instrument.
ForensicsTox_AM_PCDL.cdb
x102 Metoprolol C15H25NO3 + Product Ion Frag=150.0V CID@20.0
Scan
The instrument
serial number is recorded in the ChemStation
in the
ChemStation+ESI
ACAML
file, both of which can be
4 Metoprolol:
Are device checks used to determine, as
268.1907 full report, andx10
1
stored with the ChemStation results in OpenLAB Data Store. An OQPV must be executed
to ensure that devices and software are
appropriate, the validity
of
the
source
of
data
11.10(h)
268.1907
1.2
74.0600
116.1070
0.75
functioning properly.
or operational
instruction?
(M+H)+
98.0964
1.1
0.5
133.0648 159.0804
191.1067
User
entry fields in ChemStation
provide feedback to the user about the entry types
and ranges that are valid for a particular field.
176.1070
1
0.25
148.0757
226.1438
91.0542
250.1802
0
0.9
of the
history
are verified and can be made available during an on-site
Do the
who110
develop,
60 70
80 persons
90 100
120 maintain,
130 140 150 160 Yes.
170Records
180 190
200educational
210 220 and
230employment
240 250 260
270of Agilent employees
0.8
audit. In addition, all Agilent Technologies employees who work with regulations have attended training workshops for regulatory
or use electronic records and signature
Counts vs. Mass-to-Charge (m/z)
0.7
requirements.
systems have the education, training,
11.10(i)
0.6
andcation
experience
to perform theircan
assigned
Users of
ChemStation
/ OpenLAB
Store at atime
customer location will be required
to show records or education, training and/or
Figure 4. Identifi
of metoprolol
be further confi
rmed
by MS/MS
andData
retention
0.5
tasks?
experience
with themirror
system. spectrum,
Agilent provides
a basic familiarization during the installation of the product for system users.
matching to a reference compound (top: sample spectrum,
middle:
bottom:
0.4
269.1940
library spectrum).
0.3
Have written policies that hold individuals
(M+H)+
0.2 to develop written policies which ensure that
N/A. It is the responsibility of the organization implementing electronic signatures
accountable and responsible for actions
270.1965
individuals responsible for signing documents understand that their electronic0.1
signature is as equally binding as their handwritten
initiated under their e-signatures in order
11.10(j)
(M+H)+
signature.
to deter record and signature falsification
0
Broad Applicability
been established and followed?
268 268.5 269
269.5 270 270.5
11.10(f)
Counts vs. Mass-to-Charge (m/z)
Are there
adequate
over the
The 6550 Q-TOF
with
Jetcontrols
Stream
and iFunnelN/A.technology
delivers
theforutmost
While documentation
is available
the integration of OpenLAB CDS ChemStation Edition with OpenLAB Data Store for users
distribution of, access to, and use of
Figure
EIC of an ofunexpected
contaminant,
and administrators, controls over the storage and distribution of this material
are the3.responsibility
the organization
that implements
11.10 (k)(1) for
documentation
system of
operation
and compounds (Table 1), enabling ultra-trace
sensitivity
a broad forrange
target
and uses the system.
metoprolol, with a signal-to-noise ratio of 46:1.
maintenance?
compound identification and quantitation.
Red squares represent the theoretical isotope
Are there formal revision and change control
intensity and position.
Yes. Agilent Technologies’ quality process includes written formal revision and change control procedures for system documentation.
procedures to maintain an audit trail that
11.10 (k)(2) documents time-sequenced development and The process includes the use of OpenLAB Enterprise Content Manager (ECM) for development of system documentation. All revisions
Table 1. LOD and LOQ values (ng/L) for a broad rangeto of
compounds.
thetarget
documents
kept are time stamped and audit-trailed.
modification of systems documentation?
Compound Name
LOD
LOQ
Compound Name
LOD
LOQ
Atenolol
0.84
3.59
Sulfamethoxazole
2.97
10.84
Sotalol
0.44
2.01
Carbamazepine 10,11 epoxide
0.82
3.52
Phenacetin
0.81
3.50
11.30
Controls for Open Systems
Metronidazole
0.73
3.19
Ronidazole Question
Section
2.65
9.73
Bisoprolol
OpenLAB CDS ChemStation1.38
Edition5.52
/ OpenLAB Data Store integration
Sulfadiazine Are there procedures and2.69
controls 9.87
used
integrity11.02
and
Trimethoprim to protect the authenticity,
3.02
confidentiality of the electronic records
11.30
Sulfamerazinefrom their creation point2.55
9.39
to the point
of
Propranolol
Sulfadimidine
n,n Diethyl-3-methylbenzamide
0.95
4.02
(DEET)
Yes. The transfer of ChemStation data from its point of creation to its point of receipt in OpenLAB Data Store is fully protected. Users
their receipt?
2.59
9.53
used to ensure
Phenazone Are additional measures 0.47
2.14the
confidentiality of the electronic records from
11.30
to the point
of their
Pentoxifyllinethe point of their creation1.07
4.47
Metoprolol
receipt?
0.56
2.51
Furazolidone
2.27
8.47
Dapson
3.85
14.70
1.13
4.67
Yes. Access to all data in ChemStation / OpenLAB Data Store from both ChemStation and OpenLAB Data Store is protected using
user privileges and roles. System-generated
audit trails track all changes to electronic documents.
Carbamazepine
0.53and time-stamped
2.39
The integration of OpenLAB CDS ChemStation
with OpenLAB Data Store was not specifically designed for operation in an
Betaxolol
1.06 Edition
4.40
open system.
do not have any access to the data being transferred.
Bezafibrate
1.12
4.62
Fenofibrate
3.35
12.31
www.agilent.com/lifesciences/QTOF
The integration of OpenLAB CDS ChemStation Edition with OpenLAB Data Store was not specifically designed for operation in an
Crotamitron
0.49
2.22
open system.
For Research Use Only. Not for use in diagnostic procedures.
Diazepam
0.43
1.97
This information is subject to change without notice.
*Calculated based on the DIN 32645 using a program courtesy of Dr. Joachim Kleiner. (www.kleiner-j.de)
© Agilent Technologies, Inc. 2012
Published in the USA, April 5, 2012
5990-9862EN
271
How It Works
11.50simple
Signature
Four
stepsManifestation
with no library prep (Figure 1):
Step 1.
Question
OpenLAB CDS ChemStation Edition / OpenLAB Data Store integration
1.Section
Digest sample
DNA
Yes. The ChemStation result set or SSIZip file can be electronically signed in OpenLAB Data Store. OpenLAB Data Store’s electronic
Do the signed electronic records contain
A DNA sample
is fragmented using restriction
enzymes, and denatured.
signature manifestation includes:
information associated with the signing that
clearly indicates the following:
2.11.50(a)
Hybridize •probes
printed name of signer
• User ID in addition to the full name of the signer
Step 2.
• Date and time that the signature was applied
ate and time the signature was executed
The probe• dlibrary
is added and hybridized •to
the targeted
fragments.
Each
probeis
User-configurable
meaning
associated with
the signature
• meaning associated with the signature?
an oligonucleotide designed to hybridize to both ends of a targeted DNA restriction
these items part of any human readable
Yes. Electronic signatures applied on ChemStation files are viewable from the details page in OpenLAB Data Store. They are also
fragment,Are
thereby
guiding the targeted fragments
to form circular DNA molecules.
11.50(b)
form of the electronic record?
viewable in the OpenLAB Data Store file properties dialog in ChemStation.
The probe also contains a method-specific sequencing motif that is incorporated
Step 3.
during circularization. In addition, a sample barcode sequence is incorporated in
11.70 Signature/Record Linking
this step.
Section
Question
OpenLAB CDS ChemStation Edition / OpenLAB Data Store integration
respective electronic record to ensure
Yes. The ChemStation result set (SSIZip file) can be electronically signed in OpenLAB Data Store. The electronic Signature is
3. Purify andIs the
ligate
targets
electronic
signature linked to its
The
11.70
HaloPlex
are biotinylated
and unbreakably
the targeted
be a signature that was applied outside its own electronic
linked tofragments
the file. OpenLABcan
Datatherefore
Store will not recognize
that theprobes
signature cannot
be excised,
signature plug-in.
or otherwise transferred to falsify
retrieved copied
with
magnetic
streptavidin
beads.
The
circular
molecules
are
then
closed
an electronic record by ordinary means?
Step 4.
by ligation, a very precise reaction that ensures that only perfectly hybridized
fragments are circularized.
11.100 Electronic Signatures: General Requirements
Questionfragments with PCR
4.Section
Amplify targeted
OpenLAB CDS ChemStation Edition / OpenLAB Data Store integration
Yes. OpenLAB Data Store uses the user ID and password combination unique to each user in the electronic signature feature. User
Only circular
targets
amplified,
providing
an enriched and barcoded
Is eachDNA
electronic
signatureare
unique
to one
IDs within ChemStation / OpenLAB Data Store are required to be unique and cannot be reused or reassigned to another individual.
individual
and
not
reused
by,
or
reassigned
to,
11.100
(a)
amplification product that is ready for sequencing.
Whether ChemStation / OpenLAB Data Store uses the company’s Windows NT logins to validate users or users administrated in
anyone else?
Figure
1. HaloPlex workflow.
OpenLAB Shared Services, no two users can have the same user ID / password
combination.
Are the identities of the individuals verified
HaloPlex Delivers
Superiorassignment,
and Uniform
Coverage
N/A.
This is the responsibility of the organization that plans, implements and operates the system. Such a verification process
prior to the establishment,
and
11.100 (b)
A.
certification of an individual’s electronic
signature or any element of an electronic
signature?
11.100 (c)
Has the company delivered its corporate
electronic signature certification letter to
the FDA?
11.100 (c)
(2)
Is it in paper form with a traditional handwritten
signature?
11.100 (c)
(2)
Can additional certification or testimony be
provided so that a specific electronic signature
is the legally binding equivalent of the signer’s
handwritten signature?
is a system requirement that is set before implementing electronic signature procedures or assigning electronic signature
privileges to an individual. B.
N/A. It is the company’s responsibility, before submitting electronically signed documentation to the FDA, to register their intent
to use electronic signatures. In addition, training programs must be in place to ensure that users signing documents electronically
understand the legal significance of their electronic signature.
11.200 Electronic Signature Components and Controls
Section
Question
OpenLAB CDS ChemStation Edition/OpenLAB Data Store Integration
11.200 (a) Does the e-signature employ at least two
distinctdelivers
identification
suchuniformity,
as
Yes.and
The OpenLAB
Data Store
electronic
tool employs
identification
components:
user ID and password.
Figure 2. HaloPlex
highcomponents
specificity,
sequencing
coverage
forsignature
any design
size,two
anddistinct
across
all target
regions. unique
(A) Enrichment
of all
(i)
userCFTR
ID and password?
exons within the
gene, for 5 different samples. (B) Contiguous and highly repetitive region (~15kb) enriched with HaloPlex.
11.200 (a)
(1)(i)
When an individual executes a series of
signings during a single, continuous period of
controlled system access, is the first signing
executed using all the electronic signature
components?
Yes. When an individual signs the first of a series of documents during a single period of controlled access the user is required to
enter both signature components: user ID and password.
11.200 (a)
(1)(i)
When an individual executes a series of
signings during a single, continuous period of
controlled system access, is each subsequent
signing executed using at least one electronic
signature component that is only executable
by, and designed to be used by, the individual?
N/A. Users need to electronically sign each record individually. For eachFor
electronic
signature,
the user
enterintwo
distinct procedures.
Research
Use Only.
Nothasfortouse
diagnostic
identification components: unique user ID and password.
This information is subject to change without notice.
www.agilent.com/genomics/ngs
© Agilent Technologies, Inc. 2012
Published in the USA, April 5, 2012
5990-9855EN
How It Works
Step 1.
an individual
executes
a series
of (Figure 1):
Four simple When
steps
with no
library
prep
11.200 (a)
signings not performed during a single,
N/A. Users need to electronically sign each record individually. For each electronic signature, the user has to enter two distinct
continuous
period of controlled system
1.(1)(ii)
Digest sample
DNA
identification components: unique user ID and password.
access, does each signing executed require
all signature
A DNA sample
is components?
fragmented using restriction enzymes, and denatured.
2. Hybridize Are
probes
controls in place to ensure that only
Yes. OpenLAB Data Store can be configured such that an administrator canStep
assign2.an initial password to a user for a new account
or forgotten password, but the user is required to change that password on their first login. In this manner the user ID and password
11.200 (a)
combination is known only to the individual.
their genuine owners can use the electronic
library is added and hybridized to the targeted fragments. Each
(2)The probesignature?
probeis
Whether ChemStation / OpenLAB Data Store uses the company’s Windows NT logins to validate users or users administrated in
to
bothShared
endsServices,
of a targeted
DNA
OpenLAB
no two users can
have restric
the same tion
user ID / password combination.
an oligonucleotide designed to hybridize
fragment,Are
thereby
guiding the targeted fragments
to form circular DNA molecules.
Yes. OpenLAB Data Store applies the user ID and password to initiate the electronic signature. An OpenLAB Data Store user’s password
the electronic signatures to be
is stored
encrypted within
the database
is displayed as asterisks in all locations within the software.
and executed
to ensure that the
The probeadministered
also contains
a method-specific
sequencing
motif
that isandincorporated
attempted use of an individual’s electronic
Step installation
3.
The barcode
system administrator
is aware of user
IDs when he installs
he can force a password change during the
during cirsignature
cularization.
In
addition,
a
sample
sequence is
incorporated
in the users. During
by anyone other than its genuine
11.200 (a)
first logon. This password is only known to each user as it is defined during the first logon. See also 11.200(a)(2). The enforcement of
owner
requires
the
collaboration
of
two
or
this policy is the responsibility of the organization that operates the system. Therefore, it requires active collaboration with the purpose
(3)this step.
more individuals?
3. Purify and ligate targets
of sharing passwords to enable irregular use of another users’ identification.
OpenLAB Shared Services can be configured so that an administrator can assign an initial password to a user for a new account or
forgotten password, but the user is required to change that password on their first login.
The HaloPlex probes are biotinylated and the targeted fragments can therefore be
electronic
signaturesstreptavidin
based on biometrics
retrieved Are
with
magnetic
beads.
The circular molecules are then closed
N/A. OpenLAB Data Store does not support signatures based on biometrics
at this4.time.
11.200 (b) designed to ensure that only their genuine
Step
by ligation,
a very
precise
owners
can use
them? reaction that ensures that only perfectly hybridized
fragments are circularized.
11.300 Controls for Identification Codes/Passwords
4. Amplify targeted fragments with PCR
Section
Question
OpenLAB CDS ChemStation Edition/OpenLAB Data Store Integration
11.300 (a)
password maintained, such that no two
individuals have the same combination of
identification code and password?
Figure
1. HaloPlex
workflow.
Whether ChemStation / OpenLAB Data Store uses the company’s Windows
NT logins
to validate users
or users administrated in
OpenLAB Shared Services, no two users can have the same user ID / password combination.
Only circular DNA targets are amplified, providing an enriched and barcoded
Are controls in place to ensure the uniqueness
Yes. ChemStation / OpenLAB Data Store requires users to authenticate with user ID and password.
amplification
that is ready
of eachproduct
combined identification
code andfor sequencing.
HaloPlex Delivers Superior and Uniform Coverage
A.
11.300 (b)
Are controls in place to ensure that the
identification code and password issuance is
periodically checked, recalled, and revised?
Yes. If using Windows domain authentication, password renewal interval is configured as part of the Windows password policy setup.
The administrator can define aBtime
. frame in which passwords are periodically revised automatically. Users are prevented from reusing
passwords.
If users are administrated in OpenLAB Shared Services, OpenLAB Shared Services can be configured such that user passwords are
automatically, periodically revised.
11.300 (c)
Are there loss management procedures in
place to electronically disable lost, stolen,
missing, or otherwise potentially compromised
tokens, cards, and other devices that bear
or generate identification code or password
information?
N/A. Neither OpenLAB Data Store nor ChemStation support devices that bear or generate identification codes, such as tokens or cards, at
this time.
11.300 (d)
Are transaction safeguards in place to
prevent unauthorized use of passwords
and/or identification codes?
Yes. ChemStation / OpenLAB Data Store can be configured such that only the user knows their user ID and password. Passwords are
always displayed as asterisks and are stored encrypted within the database so that even an administrator cannot see them.
Yes. The Windows security policy can be configured so that a user defined number of unauthorized access attempts locks out the
user account and sends email notification to a system administrator. The system audit trail documents general events such as logon
attempts to the computer as well as application or user changes, in the Windows Event log as a central audit repository for all security
information. This includes the system and computer ID along with the operator name and application identification, allowing for an
immediate check of the potential security leak.
Are transaction safeguards in place to detect
and report, in an immediate and urgent
11.300 (d) manner, any attempts at their unauthorized
use to delivers
the systemhigh
security
unit and, as uniformity, and sequencing coverage for any design size, and across all target regions. (A) Enrichment of all
Figure 2. HaloPlex
specificity,
If users are administrated in OpenLAB Shared Services, OpenLAB Shared Services can be configured such that a configurable number
appropriate, to organizational management?
exons within the
CFTR gene, for 5 different samples.of(B)
Contiguous
highly
regionfor
(~15kb)
enriched
with HaloPlex.
unauthorized
accessand
attempts
lockrepetitive
out the user account
an amount
of time determined
by the administrator and logs this event to
system activity log.
11.300 (e)
Are there controls in place to initially test
devices that bear or generate identification
code or password information to ensure that
they function properly and have not been
altered in an unauthorized manner?
N/A. Neither OpenLAB Data Store nor ChemStation support devices that bear or generate identification codes, such as tokens or cards,
at this time.
www.agilent.com/genomics/ngs
For Research Usewww.agilent.com/chem/openlab
Only. Not for use in diagnostic procedures.
This information is subject to change without notice.
This information is subject to change without notice.
© Agilent Technologies, Inc. 2012
© Agilent Technologies, Inc., 2012
Published in the USA, April 5, 2012
Published in the USA, October
15, 2012
5990-9855EN
5991-1470EN