Version Approver: James Edwards Version Approval Date: 25/04/2012. Version Author: Huma Zafar (BMS) Version Date: 20/02/2012. No.

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Title: Processing of Urine Samples Serial Number: OMB-LSOP 036 Version Number: 1.0 Version Approver: James Edwards Version Approval Date: 25/04/2012 Version Effective: Two weeks after release Version Author: Huma Zafar (BMS) Version Date: 20/02/2012 Area of Application: OMB Laboratories Relevance: OMB Laboratory Staff Date Details of Review Version number No. of pages Name of Reviewer Next Review 20/02/12 New SOP 1.0 Gareth Bicknell February 2013 YOU ARE INSTRUCTED TO READ THE FOLLOWING THOROUGHLY BEFORE PROCEEDING TO UNDERTAKE THE METHODS DESCRIBED. UNDER NO CIRCUMSTANCES ARE THESE INSTRUCTIONS TO BE AMENDED OR ALTERED IN ANY WAY OTHER THAN BY THE AUTHOR / APPROVER. Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 1 of 7

Table Of Contents 1 PURPOSE... 3 2 SAFETY INFORMATION... 3 3 DEFINITIONS... 3 4 REQUIREMENTS & RESPONSIBILITIES... 3 4.1 GENERALLY... 3 4.2 SPECIFICALLY... 3 4.3 SAMPLES... 4 5 REFERENCES & RELATED DOCUMENTS... 4 6 PROCEDURE... 4 6.1 PROCESSING AND CENTRIFUGATION OF BLOOD SAMPLES... 4 6.2 HANDLING OF EXCEPTIONS... 5 7 IMPLEMENTATION... 5 8 TRAINING REQUIREMENTS... 6 9 STAFF RECORD OF ACKNOWLEDGEMENT... 7 Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 2 of 7

1 Purpose 1.1 This SOP describes the standard processing of urine samples entering the OMB. 2 Safety Information 2.1 Human tissue samples are potentially hazardous to health and should therefore be handled with care and in accordance with the policies of the University of Oxford. 2.2 Please read the associated Control of Substances Hazardous to Health (COSHH) risk assessment before using this standard operating procedure. 2.3 Policy documents are held on the University of Oxford intranet. 2.4 Working with ultra low temperatures is hazardous; therefore, all procedures should comply with local safety rules specific to these chemicals, such as liquid nitrogen. 2.5 Personal Protective Equipment must be worn at all times when handling samples. 2.6 Bench surfaces should be decontaminated with 10% Virkon solution followed by 70% industrial methylated spirit (IMS) before and after every blood preparation. 3 Definitions 3.1 Not applicable. 4 Requirements & Responsibilities 4.1 Generally 4.1.1 OMB Laboratory Staff are responsible for processing and storing samples as quickly and safely as possible, in order to ensure the highest tissue quality. 4.2 Specifically Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 3 of 7

4.2.1 Individual staff are responsible for attending appropriate training, and for attending Occupational Health. 4.2.2 The OMB Manager is responsible for ensuring that laboratory staff attend appropriate training before beginning unsupervised work, and Occupational Health before beginning any work in laboratories. 4.3 Samples 4.3.1 Samples should be collected in urine pots given on the day of the patients visit at the pre-operative assessment clinic (POAC) or from the OMB urine sample boxes kept in wards A, D E and F at NOC. The ward samples will be collected in urine pots that were given to patients on their previous visit. 5 References & Related Documents 5.1 OMB-LSOP 038 Use of the Centrifuge 5.2 OMB-LSOP 007 Use of variable volume pipettes 5.3 OMB-LSOP 002 Acceptance of Human Tissue Samples into the OMB 5.4 OMB-LSOP 013 All Waste Disposal 5.5 OMB-LSOP 014 Chemical Waste Disposal 5.6 OMB-LSOP 015 Disposal of Speciments, and Clinical Waste Disposal 5.7 OMB-LSOP 023 Laboratory Disinfection 5.8 OMB-F 003 Sample Tracker and Laboratory Checklist 5.9 OMB-F 009 Sample Tracker and Laboratory Checklist (Externally Approved Projects) 6 Procedure 6.1 Processing and Centrifugation of Blood Samples 6.1.1 If urine cannot be processed immediately, it should be stored at 4 C for up to 4 hours (or up to a time limit requested by the client project). Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 4 of 7

6.1.2 Prepare cryovials with orange inserts: Ward urine samples maximum of eight cryovials POAC urine samples maximum of five cryovials 6.1.3 Mix the urine by inverting the urine pot a couple of times. 6.1.4 If the client project requests centrifugation, transfer the urine to centrifuge tubes and spin for 10 minutes at room temperature at 1500g, brake 2. See OMB- LSOP 038 for further guidance on the use of the centrifuge. 6.1.5 Carefully use a variable volume pipette to dispense 1000 µl aliquots of urine (or supernatant, if centrifuged) into the cryovials. See OMB-LSOP 007 for further guidance. 6.1.6 Screw the lid(s) of the urine pot/centrifuge tubes back on (to prevent spillage of excess urine). 6.1.7 Snap-freeze and continue check-in of samples as per OMB-LSOP 002. 6.1.8 Waste should be disposed of according to OMB-LSOP 013-015. 6.1.9 Disinfection should be performed as per OMB-LSOP 023. 6.2 Handling of Exceptions 6.2.1 If receiving a urine pot that clearly has leaked in its specimen bag, record this on the Sample Tracker (page 1 of OMB-F 003 or OMB-F 009) and in the sample inventory system. 6.2.2 Process the sample as usual, but clean the pot surface with 70% industrial methylated spirit (IMS) and make sure it has stopped leaking before processing. 6.2.3 If no urine is retrieved, make a comment on the Laboratory Checklist (page 2 of OMB-F 003 or OMB-F 009) as well as in the inventory system. Such samples should be retired without performing an inventory Split operation. 7 Implementation Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 5 of 7

7.1 This SOP is unlikely to have direct impact on the Quality Management System, other than a requirement to know, in advance, which procedural variations are required (and to record their use in the sample inventory). 7.2 Changes to the details of this SOP may, however, affect the quality of samples from a scientific perspective. 7.3 Changes are likely to be communicated rapidly. 8 Training Requirements 8.1 Basic laboratory skills. These may be enhanced by attending courses provided by the University and/or NHS. 8.2 All new members of staff must be inducted and their competency assessed by a senior member of the team. Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 6 of 7

9 Staff Record Of Acknowledgement 9.1 I understand the contents of this document. 9.2 I have received the training appropriate to the procedures and feel competent to undertake them. 9.3 My supervisor agrees that I am able to perform the work covered by this SOP. 9.4 I understand that further on the job or other training supervision may be required before working independently. 9.5 I understand that I may discuss my needs with my manager. Trainee Trainer (if applicable) Name Position Signature Date Signature Date Approved 25/04/2012 OMB-LSOP 036 Version 1.0 Page 7 of 7