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