To verify the effect of temperature and humidity to the heparin coating / Document#2005-061
Terumo Confidential / Page 1 of 9
Report of the verification of the effect of temperature and humidity to the heparin coating

Study Number

2005-061

REPORT APPROVAL
Prepared by / Date
Jun Yamashita
R&D Engineering / Date
N/A / N/A
Manufacturing / Date
Subhan Khan
Quality and Regulatory Affairs / Date
John Toigo
  1. Purpose:

The purpose of this studywas to verify the effect of temperature and humidity to the heparin coating under the cyclic testing. The cyclic testing condition s followed the format established by IEC, and the temperature rate of change followed the EN45502.

These worst case conditions will provide the minimal amount of heparin activity while still within this range.

  1. Reference/Attachments:

2.1 The following documentation is used in conjunction with this study:

2.1.1Preparation of Heparin Solution #809653 rev. E

2.1.2 Preparation of 0.1% PEI Solution #809650 rev. E

2.1.3 Preparation of 0.03% Dextran Sulfate Solution #809651 rev. E

2.1.4 Inflow conduits/ outlet pipes heparin coating process #809657 rev. D

2.1.5Stainless Steel Disc Drawing #809684 rev. B

2.1.6Titanium DiscDrawing #809685 rev. B

2.1.7Thrombin Inhibition Assay (TIA) #812079 rev. A

2.1.8THI Sterilization Criteria Procedure #811891rev. A

2.1.9 Parts cleaning procedure# 809401 rev. D

2.1.10LAL test products and process monitoring Procedure #811177 rev. A

2.1.11 Heparin Aging study #2004-022

2.1.12 IEC 62-2-38 Basic environmental testing procedures

2.1.13 EN45502-1: 1997 AIMD General Requirements for Safety, Marking, and

Information to be provided by the manufacturer

2.2The following equipment and tooling is used for this study:

2.2.1Spatula,Spoon, Hemostats

2.2.2Disposable plastic weighing trays

2.2.3Teflon coated stirring bar p/n 811285

2.2.4250 ml Beakers, Cole-Parmer p/n A-06010-55

2.2.5800 ml Beakers, Cole-Parmer p/n A-06010-75

2.2.6Personal protective equipment (Face shields, safety glasses, nitrile gloves,

cleanroom gowns, hairnets)

2.2.7Scale,Calibration ID – 27-0026-01

2.2.8Magnetic stirrer model# PC-510 27-0032-01

2.2.9Magnetic stirrer model# S46415 27-0032-02

2.2.10Heparin temperature controller, Calibration ID – 27-0001

2.2.11Thermometer, Calibration ID – 27-0193-01

2.2.12pH meter, p/n 27-0212-01

2.2.13Coating process holding oven and thermometer 27-0031-01

2.2.14Chemical fume hood 29-0203-02

2.2.15Mixing station 27-0034-00

2.2.16Heparin hood 27-0034-01

2.2.17Ultrasonic cleanermodel# 08892-01

2.2.18LAL station model# SHKE3000

2.2.19Temperature and humidity chamber PJ-349

3. Deviations to Protocol:

The evaluation method to determine heparin activity, thrombin inhibition assay, was changed from the previous method of #78-8067-8152-8 to #812079 rev A. Therefore the acceptance criteria have changed from greater than 0.008 units/cm2 of heparin activity to the following specification.

Titanium and Stainless Steel > 0.010 U/mL

THI procedures 811891A Sterilization Release Criteria and 812079A Thrombin Inhibition Assay are in a pre- released state. Protocol indicates they have already been released. This will not effect the testing performed in this protocol.

Graph 1, 2 and 3 (General cyclic test condition cycle A, B and C) is not detailed condition.

The cyclic test was conducted as following condition.

  1. Traceability:

4.1.All the materials used during the process performance qualification are listed below.

Material / Part number / Lot number
Polyethyleneimine / 801208 / 08503CC
Dextran Sulfate / 801209 / 2324B55-100G
Sodium Chloride / 801211 / 043832
Citric Acid Monohydrate / 801210 / 040033
Sodium Hydroxide / 801212 / 031355
Sodium Cyanoborohydride / 801214 / 12807
Periodate Oxidized Heparin / 809962 / 6351007252
Stainless Steel Disc / 809684 / 66159
Titanium Disc / 809685 / 80544
Sterile 70% Isopropanol / 810182 / 3302044
Sterile water for Irrigation, USP / AB-7139-09-19 / 16-136-JT

4.2.All equipment used during the making of the samples are listed Section 2.2.

4.3.Manufacturing records and data sheets for this study are attached.

4.4.All equipment used during the making of thesesamples are listed Section 2.2.

4.4.1.Sample preparation

Sample discs were cleaned according to parts cleaning procedure.

Sample discs were strung on a wire, where each disc was separated by spacersso that the surfaces of the discs did not interfere with one another.

4.4.2.Heparin coating

Sample discs were coated according to the designated procedure under the nominal process parameter.

4.1.3LAL Extraction

Sample discs weresubmerged in USP sterile water for irrigation for one hour and thoroughly dried to simulate the LAL test.

4.1.4 Sterilization

Samples were packed in Tyvek pouches and sent for sterilization.

4.1.5 Cyclic testing

Samples were sent to Shonan R&D center in Japan, and were processed temperature and humidity cyclic test. Samples operating record is attached.

  1. Results:

Table 1: Summary of results, values for heparin activityare in U/mL unless otherwise noted.

Titanium disc / Stainless Steel disc
Non- Cyclic tested (n=10) / Cyclic tested (n=10) / Non- Cyclic tested (n=10) / Cyclic tested (n=10)
Average (U/mL) / 0.031485 / 0.019255 / 0.029004 / 0.014490
Standard Deviation / 0.001790 / 0.002988 / 0.002079 / 0.001670
Maximum / 0.033463 / 0.022690 / 0.030666 / 0.016935
Minimum / 0.027280 / 0.013450 / 0.023496 / 0.011829
Acceptance criteria / Greater than 0.01 U/mL / Greater than 0.01 U/mL

Figure 1: Effect of cyclic testing (temperature and humidity changes) on heparin activity for

Titanium substrate

Figure 2: Effect of cyclic testing (temperature and humidity changes) on heparin activity for

Stainless Steel substrate

Figure 3:Heparin activity summary and data distribution for titanium Non- Cyclic tested

Figure 4:Heparin activity summary and data distribution for titanium Cyclic tested

Figure 5: Heparin activity summary and data distribution for Stainless SteelNon-Cyclic tested

Figure 6:Heparin activity summary and data distribution for Stainless SteelCyclic tested

  1. Conclusions:

Based on the results detailed in section 5 it can be seen from figures1 and figure 2 that temperature and humidity changes have effect on heparin activity degradation.

It can alsobe seen for each substrate, that heparin activity exhibits a normal distribution(α=.05) except for Stainless Steel Non- Cyclic tested. One Stainless Steel sample of Non- Cyclic tested is lower than the others (nine data), however the heparin activity was still 130% higher than the acceptance criteria.

Review of the results detailed in section 5 indicate that for both Non - Cyclic tested and Cyclic tested samples the acceptance criteria of 0.01 U/mL for heparin activity is exceeded in all cases for both substrates.

Therefore it can be concluded that under the cyclic test, the existing heparin coating process is able to deliver heparin activity levels that can be sustained for Pump Product Requirement Specification already defined. ( Temperature 0-50 degree C, Humidity 10-95%RH)

Appendix

Raw Data