SOP for Handling of Biological Samples

OBJECTIVE
This SOP defines the procedure for collection and processing of blood samples, separation of plasma/serum from blood samples, storage and transfer of plasma/serum/whole blood samples.
 

SCOPE

This SOP is applicable to PK/PD with/and Phase I, II, III and IV studies.

 

PRECAUTIONS 

• Proper care should be taken while handling the blood sample/plasma/serum during transfer to avoid spillage. (appoint the trained Phlebotomist from Sponsor/Hospital if Applicable) 
• Correctness of the vaccutainer type and capacity and to be used for the collection as per the study protocol should be checked.

 

PROCEDURE
A. Procedure for Blood Sample Collection

• The Phlebotomist/Nursing Staff/designated person will arrange the labeled vacutainer/centrifuge tubes in the racks in each blood sample collection station as per the sampling time point.
• The Phlebotomist/Nursing staff/designated person will ensure the availability of the required medical accessories like labeled vacutainers/centrifuge tubes with appropriate anticoagulant as mentioned in the study protocol, syringes, cotton, tourniquet, vacutainer/centrifuge tube stands, needles, gloves and blood sample collection formats, or any other requirement(s) as specified in the study protocol, on the sample collection table before the start of the blood sample collection activity.
• The Phlebotomist/Nursing staff/ designated person of standby collection station will check the pre-labeled vacutainers/centrifuge tubes and will arrange them in sequence on the rack for each sample collection station and will enter the details in ‘Pre-Sample Collection Vacutainer/Tube Check format’ Intravenous cannula will be inserted in the fore arm of the subject for collection of the blood samples for Pharmacokinetic study.

• The Subject custodian will call the subject(s) by their subject number in sequence and will direct them to the respective sample collection station, at least 2 minutes before the scheduled blood sample collection time.
• Bed-side blood sample collection will be done as per requirement or if stated in the study protocol.
• The Phlebotomist/Nursing staff/designated person will verify the vaccutainer/centrifuge tube label for the study code, sampling time and the subject number and will also verify the identity of the subject by checking photograph and subject number on the Identity card and the subject number on the wrist band, before the start of each blood sample collection.
• The sample collection timings will be staggered as per the dosing time of each subject. Phlebotomist/Nursing staff/designated person will record the schedule time of each sample time point mentioned in the ‘Blood Sample Collection Details’ of the CRF for each subject.
• The Phlebotomist/Nursing staff/ designated person will discard the initial saline mixed blood (approximately 0.3 mL or volume as per Study specific protocol) from the intravenous canula using a 2 mL syringe before collecting the blood sample.
• The Phlebotomist/Nursing staff/ designated person will secure the disposable (10 mL/5 mL/2 mL) syringe to the intravenous cannula, will open the three-way stop-cock and will collect the blood sample on the scheduled time within two minutes (or as mentioned in the study protocol The volume of the blood sample to be collected will be as per the Study Protocol. After collection of blood samples, the vacutainer/tube with samples will be shaken mildly for mixing with the anticoagulant in the vaccutainer/tube.
• After collecting the blood sample, the 3-way stop cock will be closed and the blood sample will be transferred into the respective vacutainer/centrifuge tubes of the subject.
• At the time of transfer of blood samples the Phlebotomist/Nursing staff/ designated person will ensure that the samples are transferred into the correct vacutainer/centrifuge tubes of the respective subject.
• After each sample, 1 mL isotonic saline solution will be injected into the 3-way stop cock to avoid blockage of cannula. The 3-way stop cock will be kept closed till the next scheduled sample collection of the subject.
• The Phlebotomist/Nursing staff/ designated person will record the actual time of each sampling time point for each subject and will sign and date in the format for ‘Blood Sample Collection Details’ of the CRF.
• In case of cannula blockage observed during the sample collection, the blood sample will be collected by direct vein puncture using a syringe with needle, thereafter the subject can be recanulate, if required. Any deviation in the sample collection time of more than two minutes (or as mentioned in the study protocol) and the reason for the same (canula block/canula replaced/direct prick/poor blood flow) will be recorded in the ‘Blood Sample Collection Details’ of the CRF of respective subject.
• The Phlebotomist/Nursing staff/designated person will transfer the vacutainer/centrifuge tubes with blood samples at each sampling time from each sampling station(s) to the sample processing area.

• The Technician/designated person will receive the vacutainer/centrifuge tubes with blood samples and check for the time point, receiving time, total number of samples received, and record the details in ‘Centrifugation and Separation of Biological Samples Record’.
• In case of requirement of whole blood or serum, the sample will be processed as specified in the respective Study Protocol.
• The Technician/designated person will place the vacutainer/centrifuge tubes in the centrifuge machine.
• The Technician/designated person will set the speed (RPM), time and temperature (as mentioned in the study protocol) of centrifuge machine and will operate the centrifuge machine as per the Standard operating procedure for Centrifuge machine. The details of the set parameters and the centrifugation start and end time will be recorded in ‘Centrifugation and Separation of Biological Samples Record’.
• After completion of the centrifugation, the Technician/designated person will remove the vacutainer/centrifuge tubes carefully (without disturbing the contents) from the centrifuge machine and will arrange it in vacutainer/centrifuge tube stand.
• The Technician/designated person will arrange the pre labeled vials of respective sampling hour sequentially in the rack/tray.
• The Technician/designated person will harvest the plasma/serum from each centrifuged vacutainer/centrifuge tube using micro pipette with disposal tips without disturbing the sediment layer and will transfer the harvested plasma/serum to respective subject’s pre labeled vials into two aliquots as ‘Replicate’ (1 mL) and ‘Analytical’ (remaining quantity) sample. Or as specified in the Study Protocol, and will cap the vials.
• The Technician/designated person will record the observation [like hemolysed samples (H), missing sample (M), subject not reported (N)], or any other observation as remark in the ‘Centrifugation and Separation of Biological Samples Record’.
• The Technician/designated person will pack these vials with plasma/serum samples of each sampling time point into labeled zip-lock bags (each for Analytical and Replicate samples) of all the subjects, and store in the deep freezer maintained at suitable storage temperature as specified in Study Protocol.
• The Technician/designated person will record details of samples stored in the deep freezer.

 

 
B. Precaution to be taken for Photo sensitive and Temperature Sensitive Drug Product.

• For study of Photosensitive molecules, appropriate measures will be taken (like use of sodium vapor lamp/covering the glass windows with dark paper) to avoid exposure to light while performing all the procedures done in sample collection and processing area.
• The Technician/designated person will use pre labeled amber colored vials for storage of Plasma/Serum samples of light sensitive drug product.
• For temperature sensitive drug products, the following measures will be taken or will follow the measures/procedures mentioned in the respective Study Protocol.
• The blood sample will be collected into pre-chilled vacutainer /centrifugation tube. This vacutainer/centrifugation tube with blood samples will be placed in ice water bath till it is centrifuged. After centrifugation at the set parameters the plasma/serum will be harvested using micropipettes with disposable tips into pre-labeled vials as
• ‘Analytical’ and ‘Replicate’ sample, these vials containing the harvested plasma/serum will be placed into deep-freezer maintained at a temperature specified in the Study Protocol.
• The Phlebotomist/Nursing staff/designated person will record the sample time point deviation in the ‘Deviation Reporting Form’. The responsible Physician/designated person will also give the details of sample time point deviation, missing samples and the delay in sample time for ambulatory samples to the Statistical Department for considering in the statistical analysis.

 

 
C. Transfer of samples from the Clinical Facility to the Bio-analytical Facility.

• Study Coordinator/designated person will take the authorization from Clinical investigator/designated person for transfer of biological samples to the bio-analytical facility and will get the name and address of the bio-analytical facility where the sample has to be transferred.
• Study Coordinator/designated person or the Contract Courier person (who will be shipping the samples) will arrange insulated boxes with appropriate coolant before initiation of the transfer activity.
• Study Coordinator/designated person or the Contract Courier person will place the temperature monitoring device to record the temperature during the transfer of biological samples.
• Study Coordinator/designated person will identify and remove the biological sample boxes/polybags from the deep-freezer and will record the retrieval of samples in respective format.
• Study Coordinator/designated person will transfer these labeled samples
• boxes/polybags as per the specified requirements, into the insulated box with sufficient coolant to maintain the storage condition of the biological sample during transportation and will seal the box (es).
• Study Coordinator/designated person will record the details of samples to be transferred in the format for ‘Transfer of Biological Samples from Clinical facility to Bio-analytical Facility’.
• Study Coordinator/designated person or the Contract Courier service person will paste appropriate labels {e.g. warning label as “Biological Samples”, “Handle with Care”, “Light Sensitive samples” (if any)} and will also paste label mentioning the complete address of the bio-analytical facility where samples are to be transferred, on the box. If Applicable.
• Study Coordinator/designated person will handover the samples/insulated box(es), the format for ‘Transfer of Biological Samples from Clinical facility to Bio-analytical Facility’ and the covering letter with the details of the samples, to Contract Courier service person or the person shipping the samples (in case the samples are hand delivered).
• The Contract Courier service person will sign the transfer format as ‘Received by’ and will write the time of receipt. The Contract Courier person will also fill the details in the shipment tracking form, and submit a copy of it to the Study Coordinator/designated person.
• In case the samples are hand delivered the person receiving the sample at bio-analytical facility will verify details of the sample from the covering letter and/or from the transfer format and sign as ‘Received by’ write the time of receipt on the format and will acknowledge the receipt of the samples.
• A copy of this transfer format will be maintained in the Study File at the Clinical site.
• The person receiving the sample at bio-analytical facility/Contract Courier service person will remove the temperature monitoring device and the print of the temperature record during transportation will be maintained along with the Sample transfer format. If it is genetic samples (Please submit the Material transfer agreement and Data Transfer Agreement to the Institution/IEC.

ANNEXURE
Nil

REVISION HISTORY
Nil

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