Fluid
Technology
Solutions

FULL VALIDATION GUIDE

Pumpsil® Issue 12

Executive Summary
This is the full validation guide for Pumpsil tubing. It contains
additional hyperlinks to the test reports. The additional
hyperlinks are provided in section 8 page 10. This should
provide the reader with detailed test procedures and actual
test results.

CONTENTS

1 Introduction
2 Irradiation and autoclave conditions and working temperature
3 Chemical compatibility
4 Regulatory compliance statement and manufacturing conditions
a.
b.
c.
d.
e.
f.
g.

Materials of construction
Manufacturing environment
Country of origin
Compliance declaration summary
REACH legislation
RoHS
Shelf life storage conditions

5 Biocompatibility and physiochemical testing
a.
b.
c.
d.
e.
f.
g.
h.
i.
j.
k.
l.
m.

Summary table
USP <88>
USP <87>
ISO 10993-4
ISO 10993-6
Kligman maximisation test – ISO 10993-10
ISO 10993-5
USP<381>
ISO 10993-11
ISO 10993-10
USP <85>
USP <788>
European Pharmaceopeia 3.1.9

6 Extractables testing
7 Conclusions
8 Downloadable validation document links
11. Introduction
Watson-Marlow’s Pumpsil 913.A and 913.B series platinumcured silicone tubing is a high purity, ‘addition cured’
silicone which is USP Class VI and ISO 10993 compliant,and produced from a USP Class VI raw material.
It is manufactured in an ISO 14644-1 class 7 cleanroom.
The cleanroom is temperature and humidity controlled,thereby providing a stable extrusion environment.
Pumpsil 913.A and 913.B series platinum-cured silicone
tubing exhibits a number of key features:• Ultra-smooth bore to control protein binding 		and bacterial growth. Tight surface cure.
Non-tacky surface• Traceable with laser etching of the part 			number, lot number and use-by date along the 		length of the tube. Full traceability is therefore 		assured even when the tubing is removed from thebag• Excellent flow stability for accurate process 		
control• Very low extractables, non-toxic and Animal 		
Derived Content Free (ADCF)

Pumpsil is fully post-cured, a process which removes cyclic
siloxanes and other volatiles, resulting in an exceptionallypure tube. Another major benefit of post-curing is increased
cross-linking between molecules. These tighter molecularbonds maximise resistance to flexing stress. The tubing
fatigues more slowly and therefore retains its original
shape for longer and dispensed volumes remain accurate
throughout the tube life.
Watson-Marlow Fluid Technology Solutions (WMFTS) is theworld leader in niche peristaltic and sinusoidal pumps andassociated fluid path technologies. Founded on nearly 60
years of supplying engineering and process expertise andwith over one million pumps installed worldwide, our pumps
are tried, tested and proven to deliver.
WMFTS is a wholly owned subsidiary of Spirax-SarcoEngineering plc (LSE: SPX), a global organisation employing
approximately 4,800 people worldwide. Watson-Marlow
Fluid Technology Solutions comprises nine establishedbrands, each with their own area of expertise, but together
offering our customers an unrivalled breadth of solutions fortheir pumping applications.

• Comprehensive stock of a wide range of sizes22. Irradiation and autoclave conditions and working temperature
Pumpsil tubing may be sterilised using either of the following
methods:• Gamma irradiated to 50kGy• Autoclaved at 121C for up to 60 minutes

3. Chemical compatibility
A general guide on chemical compatibility of Pumpsil
tubing can be found on Watson-Marlow Fluid Technologies
Solutions website. wmfts.com/chemical

3

Working temperature range• -20C to +80C
The working temperature range for Pumpsil 913.A and
913.B series silicone tubing has been determined based onits use as a peristaltic tube in Watson-Marlow pumps.
For static (i.e. non-pumped) applications these limits may be
exceeded, but it is recommended that testing is carried out
to ensure suitability.

4. Regulation compliance statements and manufacturing conditions
4a		

Materials of construction

4e		

Pumpsil is made of polydimethylsiloxane (PDMS)
structure with a high surface area silica filler toprovide enhanced physical characteristics. The
tube contains less than 1% catalyst residue, whichitself is primarily a silicone carrier for the platinum
catalyst. Platinum is not detected in the aqueous
extracts from the finished tube (see section 6).

4b		

All raw materials, compounds used in the
manufacturing process and the final Accusil
product comply with the REACH regulation. Noneof the chemicals used in the manufacture of Accusil
are on the candidate list of substances of 2008 or
the list of Substance of Very High Concern (SVHC).

4f		

Manufacturing environment

4g		

Country of origin

Shelf life storage conditions
The use-by date of the tubing is included on the
bag and box label. To maintain the performanceof the tubing throughout its life, tubing should
be stored in a cool, dry environment away fromdirect sunlight. The optimum storage temperature
is between 18-21C (65-70F). However, normal
warehouse conditions of 5-30C (40-86F) are
acceptable. Wherever possible, original packaging
should be maintained. Stock should be rotated ona first in, first out (FIFO) basis.

Pumpsil is manufactured in Falmouth, Cornwall,
United Kingdom.

4d		

RoHS
In compliance with the restriction of hazardous
substances (RoHS) directives, no listed substances
are used in the manufacture of Accusil.

Pumpsil is manufactured according to current Good
Manufacturing Practices (cGMP) in an ISO 14644-1
Class 7 cleanroom.

4c		

REACH legislation

Compliance declaration summary
Table 1 details the different substances that are notpresent in the raw material, manufacturing process
or final composition of Pumpsil.
For full compliance statements please refer to thecompliance summary sheet available from thewebsite or WMFTS representative.

The performance of any tubing beyond its use-bydate, or which has not been stored according to
the recommendations outlined above, cannot be
assured.

Table 1: List of compliance statements for Pumpsil and substances not present in the manufacture of Pumpsil
Named substance/compliance statement

Raw material

Manufacturing process

Final product

Gluten---Animal Derived Content Free (ADCF)

-

-

-

Melamine---

Phthalates

-

-

-

Bisphenol A (BPA)

-

-

-

Latex

-

-

-

Allergens (as defined by FDA CFR 21.164.110)

-

-

-

Presence of heavy metals

-

-

-

‘-’ denotes not present or not added

4

5. Biocompatibility and physiochemical testing
5a		

Summary table
Table 2 contains a summary of all the compendial
testing and ISO qualifications that Pumpsil has
been evaluated for. Full test methods and results
are available on request.

Pumpsil has passed a number of compendial and ISO
testing, a summary of the results are disclosed within.

Table 2: A summary of the compendial and ISO tests performed on Pumpsil
Test reference

5

Test description

ResultISO 10993-11

Systemic toxicity

PASS

ISO 10993-10

Intracutaneous injection – test for irritation and skin sensitization

PASS

EP 3.1.9

European Pharmacopoeia 3.1.9 silicon elastomer for closures andtubingPASS

ISO 10993-4

Hemolysis test – Autian method

PASS

ISO 10993-10

Kligman maximisation – Test for irritation and delayed typehypersensitivity

PASS

USP <87>

In vitro cytotoxicity testPASS

ISO 10993-5

Quantitative MEM elution– ISO. Biological reactivity part 5, in vitro
cytotoxicity

PASS

USP <381>

Physicochemical tests on elastomeric closure materials

PASS

ISO 10993-6

Short term intramuscular test

PASS

USP <88>

USP Class VI summary certificate

PASS

USP <85>

Limulus Amebocyte Lysate (LAL) Bacterial Endotoxin Assay

REPORT

USP <788>

USP Particulate/ Microscopic particulate count analysis test

REPORT5b USP <88> Biological Reactivity Tests,
		 In Vivo, Post Gamma Irradiation 					 samples

5d

The hemolysis test assesses the potential forindirect contact of a given sample with blood tocause the rupture of erythrocytes (red blood cells).

USP Class VI Plastics Test assesses the potential
toxicity of a given test article systemically,
intracutaneously and through implantation.

Pumpsil were tested in accordance with
ISO 10993-4, 2002, Biological Evaluation of
Medical Devices – Part 4: Selection of Tests forInteractions with Blood.

Samples of the Pumpsil were gamma irradiatedat 45 – 55kGY and tested in accordance with
USP32, NF 27, <88>, biological reactivity tests,
In Vivo. This included the immersion of the test
articles in the following solutions: USP 0.9% NaCl,cottonseed oil, 1 in 20 ethanol in NaCl solution andpolyethylene glycol 400 at 70C for 24 hours.
Results: Pumpsil extracts and implants
showed no toxicity, therefore Pumpsil passed
USP Class VI testing.

5c		
USP <87> Biological Reactivity Tests, In
		 Vitro, Post Gamma Irradiation samples
USP Biological reactivity tests, In Vitro, poststerilisation samples Samples of Pumpsil were
gamma irradiated at 45–55kGy and tested inaccordance with USP43, NF38, , Biologicalreactivity tests, In Vitro. The biological reactivity ofa mammalian monolayer, L929 mouse fibroblast
cell culture in response to Pumpsil was determined.
Samples of Pumpsil tubing, positive control (rubber)and negative control articles were prepared at 37°C
for 48 hours. Biological reactivity was rated on ascale ranging from Grade 0 (no reactivity) toGrade 4 (severe reactivity).

ISO 10993-4 Hemolysis

Results: Per ISO 10993-4, Pumpsil isconsidered non-hemolytic.

5e		
		

ISO 10993-6 Short term Intramuscular 		
Implantation Test
The purpose of this test is to evaluate the solidmaterial in direct contact with living tissue.
Pumpsil was tested in accordance with
ISO 10993-6. Samples of Pumpsil (1mm x
1mm x 10mm) and the negative control plastics
were evaluated. The test sites were examined
for inflammation, encapsulation, necrosis,
haemorrhage and discolouration macroscopically.
Results: Pumpsil did not demonstrate
any difference as compared to the control
implant sites when in contact with tissue for
two weeks. Therefore Pumpsil passed therequirements for ISO 10993-6.

Results: No reactivity (grade 0) was exhibitedby the cell cultures when exposed to Pumpsil.
Therefore Pumpsil is not cytotoxic andpassed the requirements of USP biologicalreactivity tests.

6

5f		
ISO 10993-10 Kligman Maximisation 				 Testmaintenance medium on the cell cultures was
replaced by extracts of Pumpsil, or control article.
The cell cultures were incubated for 48 hours at
37C ±1°C. Biological reactivity was evaluated by aphoto spectrometer at 450nm wavelength.

		 The purpose of this test is to detect the allergenic 				potential of a test article.

Pumpsil was tested in accordance with
ISO 10993-10. Samples of Pumpsil were extracted
in USP 0.9% NaCl for injection and cottonseed oil
at 70C for 24 hours and then injected intradermally.
After two weeks, an additional topical application
was introduced to the site of intradermal injections.
Results: The sites that were exposed to thetest articles and negative control showedno signs of erythema or edema. Therefore,
Pumpsil is deemed not to contain any allergy.

Results: Pumpsil showed no signs ofcytotoxic activity. Therefore Pumpsil passed
the requirements of ISO 10993-5.

5h		
		

5g		
ISO 10993-5 Biological evaluation of 				
medical devices- part 5: tests for in vitro
		 cytotoxicity

USP <381> Elastomeric closures for 		
injections - physiochemical tests
Extracts of Pumpsil were prepared according to therequirements of USP 32, NF 27, Chapter 381 asdirected under physiochemical tests. The results ofthe tests are summarised in Table 3.
Results: Based on the evaluation criteria
mentioned below, Pumpsil tubing meetsthe requirements of the USP <381> section
physiochemical tests.

SO 10993-5 Biological evaluation of medical		devices—tests for In Vitro cytotoxicity. The
biological reactivity of a cell culture, in response
to extracts from Pumpsil was determined. The
Table 3: List of physiochemical tests that form USP <381>
Test

7

Test Result

Evaluation Criteria (Type One closures)

ResultAppearance ofsolution

0.09NTU< 6NTU< less intense than matching fluid O

PASS

Acidity or alkalinity

0.1ml NaOH to produce blue colour< 0.3ml NaOH to produce a blue
colourPASS

Heavy metals

Less intense colour than standard solution

Standard solution containing 2ppm
Lead (Pb)PASS

Reducing
substances

0.2ml

< 3ml

PASS

Absorbance

0.1763ppm< 0.2ppm

PASS

Extractable Zinc

0.0021ppm< 5ppmPASS

AmmoniumLess intense colour than standard solution

Standard solution not more than 2ppm
PASS
of Ammonium present.

Volatile Sulfides

No black stain

Less intense colour than standard
solution

PASS

5i		
		

ISO 10993-11 guidelines for the 			
Systemic Injection test

5k		
		

The purpose of the systemic injection study is
to screen test articles extracts for potential toxic
effects as a result of a single dose systemic
injection. Samples of Pumpsil tubing wereextracted using 0.9% sodium chloride for injection,
cottonseed oil, 1 in 20 ethanol in NaCl solution orpolyethylene glycol 400 at 70C for 24 hours.

USP Limulus amebocyte lysate (LAL) bacterial
entoxin assay Endotoxins are lipopolysaccharide
complexes in gram negative bacterial cell walls.
The limulus ameobocyte lysate (LAL) Kinetic
Chromogenic assay is used to determine thequantitative level of endotoxin present within our
products. Pumpsil tubing was tested in accordance
to the requirements of USP 85. Pumpsil (22.0
cm² surface area) was extracted in 50mL of
LAL reagent water at room temperature for 60
minutes. LAL-lysate was added to each sample
well on an automated microtiter plate and thentransferred to a plate reader where the absorbanceof each well was read at 405 nm and 37 °C at set
timepoints. A standard curve was generated using
software based on reaction time versus endotoxin
concentration which was then compared to theendotoxin concentration and reaction time of thesample extract.

Results: Pumpsil meet the requirements of
ISO 10993-11 guidelines for the systemic
injection test.

5j		
		

USP <85> Limulus amebocyte lysate 		(LAL) bacterial endotoxin assay

ISO 10993-10 guidelines for 			
intracutaneous Injection test
The intracutaneous test is designed to evaluate
local responses to the extracts of Pumpsil following
intracutaneous injection. Pumpsil tubing is
extracted using 0.9% NaCl for injection, cottonseedoil, 1 in 20 ethanol in NaCl solution or polyethylene
glycol 400 at 70C for 24 hours.

Results: Accusil extracts had an EU/ml valueof <0.011 EU/cm² which is comparable to thelevels of endotoxin observed in water
for injection.

Results: Pumpsil meet the requirementsof ISO 10993-10 guidelines for the
intracutaneous injection test.

5l		
		

USP <788> Particulate testing/			
microscopic particulate count analysis
testThis test is used to determine a level of particulates
measuring 10 micron (µm) or less and25 micron (µm) that may be present in any given
drug product.
A length of Pumpsil tubing was filled with low
particulate water and shaken 20 times. The
extraction fluid was then recovered and theparticles were measured using light obscuration.
Results: Extracts from a Pumpsil tube
contained 1187 particles > 10µm and 8
particles > 25µm

85m

European Pharmacopoeia 3.1.9

Results: Based on the results of the tests,
Pumpsil tubing meets the requirements of EP
3.1.9 section Physiochemical tests.

Extracts of Pumpsil were prepared in accordancewith the requirements of European Pharmacopoeia,
2009, Chapter 3.1.9 Silicone elastomer for closuresand tubing. The results of tests are summarised inTable 4.

Table 4: List of physiochemical tests that form EP 3.1.9- Silicone elastomer for closures and tubing
Test
Appearance ofsolution

Test result

Evaluation criteria

ResultClear, 1.344NTU

Clear,
Turbidity < RS I (3NTU)

PASS

2.5ml 0.01M NaOH (blue colour)≤ 2.5ml of 0.01M NaOH change to blue

Acidity or alkalinity
1.0ml 0.01M HCl (yellow changes toorange)9≤ 0ml of 0.01M HCl change to yellow toorangePASS

Relative density

1.16 g/ml

1.05 – 1.25 g/ml

PASS

Reducing
substances

0.8ml

Diff. between sample and blank ≤ 1.0ml

PASS

Substances
0.32%
soluble in hexanes

3%

PASS

Volatile matter

0.17%< 2 % ( Pt cured)

PASS

Mineral oils

Less fluorescence than 1ppm
standard

Less fluorescence than 1ppm standard

PASS

Phenylated
compounds

Max absorbance < 0.009A.u.

Absorbance < 0.4A.u. (between 250 and
340nm)

PASS

PlatinumLess coloured than Pt reference(30ppm)Less coloured than Pt reference (30ppm)

PASS6. Extractables testing

7. Conclusions

Sections of Pumpsil tubing were subjected to extraction
in multiple solvents at controlled temperatures. The test
materials was extracted in a 6cm2 : 1ml surface area tovolume ratio. The solvent extracts were then analysed forvolatile, semi volatile, non-volatile and metallic extractables
using High pressure liquid chromatography – diode array
detector - mass spectrometry (HPLC-DAD/MS), Direct
injection gas chromatography – mass spectrometry (DI-GC/
MS), Headspace gas chromatography – mass spectrometry(HS-GC/MS) and Inductively coupled plasma – mass
spectrometry (ICP/MS).

Pumpsil has been shown to pass a number ofcompendial and ISO testing summarised in this guide.
For further information with full compliance statements
and the test reports, please contact your WMFTS
representative.

33 elements were evaluated by ICP MS including thoselisted in ICH Q3D and USP 232 guidelines.
Results: These studies have shown that there are
low levels of extractables from the tubing extracts of
0.01N NaOH, 0.01M HCl and WFI solutions. WMFTS
can provide further information and assistance in theevaluation of extractables data for risk assessment
purposes.

8. Downloadable validation document links
Section Number

Title of test

Link to the report (Click hyperlink to open)

ISO 10993-11/ISO 10993-10 – Systemic
toxicity and intracutaneous injection
USP<88> Biological reactivity tests, in vivo

USP Class VI & ISO 10993 parts 6, 10 & 11

5b/5e/5i/5j

5c/5g

USP<87> Biological reactivity tests, in vitro
ISO 10993-5 – Quantitative MEM Elution in vitro
cytotoxicity

USP 87/ISO 10993-5 Qualitative MEM
ISO 10993-5 Quantitative MEM

5d

ISO10993-4 Hemolysis

ISO 10993-4

5f

ISO10993-10 Kligman Maximisation

ISO 10993-part 10 ( Kligman Maximisation)5h

USP <381> Elastomeric closures for injections

USP 3815k

USP <85> Limulus ameobocyte lysate ( LAL)
bacterial endotoxin assay

USP 855l

USP <788> Particulate testing

USP 7885m

European Pharmacopoeia 6.8, Chapter 3.1.9

EP 3.1.9

6

Extractables testing

Extractables

10FluidTechnology
Solutions

BIOTECHNOLOGY AND PHARMACEUTICAL SOLUTIONS

Watson-Marlow Fluid Technology Solutions
Watson-Marlow Fluid Technology Solutions supports
its customers locally through an extensive global
network of direct sales operations and distributors

Copyright © 2022 Watson-Marlow Fluid Technology Solutions
PUMPSIL FULL VALIDATION GUIDE issue 12

wmfts.com/global

Disclaimer: The information contained in this document is believed to be correct but Watson-Marlow Limited
accepts no liability for any errors it contains and reserves the right to alter specifications without notice.
It is the users responsibility to ensure product suitability for use within their application. Watson-Marlow,
LaserTraceability and Pumpsil are trademarks of Watson-Marlow Limited.

11

A Spirax-Sarco Engineering plc company