Rouging in pharmaceutical

water system
Per-Åke Olsson, Alfa Laval
Webinar, ISPE Singapore affiliate, 16 Feb, 2022
Rouging in water system
Introduction of speaker
Per-Åke Olsson
Global Industry Manager, Pharma & Biotech, Alfa Laval

Alfa Laval AB, 2002- (Lund, Shanghai, Lund)

Astra Zeneca R&D Lund, Sweden, 1997-2002
NiMe Hydrid AB, Mönsterås, Sweden, 1993-1997

Member of the Pharmaceutical Technology Europe editorial advisory board between 2006 and 2008,
and has been a speaker and chairman at several BioPharma conferences and seminars in Asia,
America and Europe

MSc, Mechanical Engineering, University of Lund, Sweden 1987-1993

eMBA, University of Warwick, UK 2000-2004

telephone: +46 722 171 226

email: [email protected].
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Rouging in pharmaceutical water system

• Water storage & distribution loops

• Introduction to rouge
• Regulatory stance
• Rouge formation
▪ and how to avoid or minimize risk of rouging
• De-rouging
▪ Recommendations on how to remove rouging
• What does guidelines say?

Objective is to give you a brief understanding of the phenomenon of Rouging

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Water storage & distribution loops

Water Feed water Pre treatment Final treatment

Generation

Storage &
Distribution

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USP water specification

WFI PW
Conductivity µS/cm @ 25 °C* 1,3 1,3 * Conductivity levels depending on measured temperature

TOC, ppb < 500 < 500

Bacteria, cfu/100ml 10 10 000** ** Purified water specified as 100 cfu/ml

Endotoxin, max 0,25 EU/ml Not spec.

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Rouge

Rouging is the formations of iron oxide/ hydroxide originating from

external sources due to destruction of the passive layer.
Class I Rouge
Migratory rouge

These forms of iron oxides are not soluble in blood and may cause
Class II Rouge
clogging if introduced to the blood system. In situ oxidation of non-passive surfaces

Rouge is also a good growing ground for bacteria and biofilm. Class III Rouge
Black oxide produced by hot oxidation
However, regulatory authorities has no written position directly
addressing rouging.

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Rouge classification

Class I: ”Limonite”, FeO (OH), iron / ferrous oxide

▪ Rouge that originates from up-stream sources;
migrating;non-adherent. Reddish / orange in color
Class II: Hematite, Fe2O3, iron / ferric oxide
▪ Rouge that forms in place, within the affected system; result
of damaged surface; chloride or halide attack; Integral with
the surface; more frequent on machanically polished
surfaces.
Spectrum of orange, red, blue, purple, grey, and black
Class III: Magnetite, Fe3O4, iron / ferrosoferric oxide
▪ Rouge that forms in high temperature steam systems.
Extremly stable form; reformulate after acid removal.
Gold, blue, or black and ”powdery” or glossy in appearance

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Poll 1 & 2
1. Do you have rouge in your facility?
▪ No
▪ Yes
▪ Don’t know

2. If you have rouge in your facility, which class is it? (multiple choices)
▪ Class I
▪ Class II
▪ Class III
▪ Don’t know

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ISPE
Baseline guide, Volume 4, Water and steam systems [Ref. 3]

Chapter 10, Rouge and stainless steel

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Rouging Guidance - ISPE

10.2 Regulatory stance:

Concern should be focused on whether or not the presence of rouge may be detrimental to the drug
products, rather than its repercussions on capital equipment protection.
The FDA has no written position addressing rouging, its existence, or presence in high purity water, steam,
and process systems. Their criterion is to meet established standards of quality for those systems.
21 CFR (Code of Federal Regulations) Chapter I, Part 211, Subpart D – Equipment,
Section 211.65(a) – Equipment construction.
….shall not be reactive, additive, or absorptive so as to alter the safety, identity, strength, quality, or purity
of the drug product beyond the official or other established requirements.”
Section 211.67(a) – Equipment cleaning and maintenance.
Equipment shall be cleaned, maintained……..
The USP neither identifies rouge as a contaminant nor proposes alert and action limits or methods for
detecting rouge product streams; the US Pharmacopeia usually does not address design or material
criteria directly, but rather indirectly by defining limits for the components that ultimately will enter the
human body

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USP <788> & <789> [Ref. 4]

• No visible particles allowed

• Limits for sub-visible particles in (787), 788 & 789

<788> Particulate matter in injections <789> Particulate matter in ophthalmic solutions

Test 1.A — Solutions for parenteral infusion or
solutions for injection supplied in containers with a
nominal content of more than 100 mL.

Diameter Diameter
≥ 10 μm ≥ 25 μm ≥ 10 μm ≥ 25 μm
Number of 25 per mL 3 per mL Number of 50 per mL 5 per mL
particles particles

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Rouging Guidance - ISPE
10. Rouge and Stainless steel

10.4 Rouge Formation:

………… For further details on these classifications, methods to remediate the presence of rouge
(de-rouging, and considerations that affect the amount of rouge formation during fabrication and
operation of a system, refer to ASME BPE

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Poll 3
3. Does your company have rouge guidances/limitations for water systems?
▪ No
▪ Yes
▪ Don’t know

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Rouge Formation

Stable system
Cr2O3

Class I formation

Cr2O3

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Passive layer - protective

10-50 nm i.e 0.01-0.05 µm Metal oxide (Cr2O3)

or 100-500 Å

Alloy (Fe, Cr, Ni, Mo, etc.)

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Passivation Guidance
ISPE & ASME BPE

ISPE [Ref. 3] – Some of the most common passivation treatments include the use of nitric
acid, phosphoric acid and phosphoric acid blends, ammoniated citric acid, and mixed chelant
systems. While passivation helps to build a good, thick passive layer on the product contact
surface, however, it has been observed that after the system is put back on service, the
passive layer will be the same that corresponds to the natural interaction between product and
product contact surface, making passivation a short time effect (Reference 30, Appendix 1),
and showing no general benefit to the corrosion resistance of the base metal.
ASME BPE [Ref. 5] – Process contact surfaces of austenitic stainless steel, except those
covered by section SD-4.3.1 (Process gases), should be passivated before being placed in
service.
Passivation of electropolished surfaces is not required unless the surface has been altered
(e.g., welded or mechanically polished) or exposed to external contamination after
electropolishing.

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 ASME BPE passivation requirements

NONMANDATORY APPENDIX E NONMANDATORY APPENDIX H

PASSIVATION PROCEDURE QUALIFICATION ELECTROPOLISHING PROCEDURE QUALIFICATION

2019 ASME BPE Table D-2-1 & D-2-2

Standard Copyright © 2019 by The
American Society of Mechanical Engineers

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Causes of rouge
Rouging exposure tests, Mathiesen et. al. [Ref. 1]

Atmosphere:
Atmospheric air show less rouging, CO2, N2 and other
halides show increased rouging

Materials:
No difference between different stainless steel alloys

Polishing:
Higher degree polishing, especially EP show less rouging

δ-ferrite:
Different δ-ferrite content did not show any difference on
rouging (but erosion effects)

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Causes of Rouge

Material
- Sulfur content for welding (pitting corrosion)
- Chromium content (passive layer)
- (Low delta ferrite to minimise erosion)
Fabrication
- Machining (destroy the passive layer)
- Polishing (surface contamination)
- Welding (galvanic & intergranular corrosion)
Process Environment
- Systems with CO2 ,N2 , Chlorides, or other halides
- Elevated temperature (>60 °C)
- System with physical stress and or erosion

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Steel grades

Material Grade Chemical composition (wt %, balance Fe)

Material C Si Mn P S N
number Standard max. max. max. max. max. max. Cr Ni Mo
1.4404 DIN-EN 10088-1 0,030 1,00 2,00 0,045 0,015b) 0,11 16,5-18,5 10,0-13,0 2,00-2,50
1.4435 DIN-EN 10088-1 0,030 1,00 2,00 0,045 0,015b) 0,11 17,0-19,0 12,5-15,0 2,50-3,00
316L1) ASTM A 270 0,035B) 1,00 2,00 0,045 0,030 - 16,0-18,0 10,0-14,0 2,00-3,00
0,005 -
316L1) 2) ASTM A 270 S2 0,035B) 1,00 2,00 0,045 0,017 - 16,0-18,0 10,0-14,0 2,00-3,00

1) UNS designation S31603 in accordance to ASTM A 270

2) In accordance to ASTM A 270 supplementary requirements S2 for Pharmaceutical quality tubing

Impurities! Not controlled by material standards

– Contamination, welding problems, corrosions, etc.

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Sulphur content

A low sulphur content (0,0% - 0,005%) gives a wide weld pool and it is difficult
to get a full penetration of the base material.

A high sulphur content (0,020% - 0,030%) gives micron-size pits after electro
polishing and can cause pitting corrosion (Sulphur and Manganese form
manganese sulphide inclusions)

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Surface Guidance
ISPE & ASME BPE

ISPE [Ref 3] – It is common industry practice to use sanitary tube OD sized materials for high
purity water applications with interior finishes ranging from improved finish of 30 microinch (0.8
micrometer) Ra to mirror-like surface finishes of better than 10 microinch (0.3 micrometer) Ra.
After mechanically polishing or electropolishing, the polishing compounds should be confirmed to
have been completely removed, so as not to accelerate corrosion, or contaminate the water.
Though the value of high quality finishes is undecided, finishes in the range of 25 microinch (0.6
micrometer) Ra are most common.
Systems operating at ambient temperature or with infrequent sanitization may require a smoother
surface finish.
ASME BPE [Ref. 5] – When compendial water systems are constructed of metallic materials, the
surface finish should be less than or equal to 25 µin. Ra or 0.6 µm (see Part SF) and may be
internally electropolished.

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 Product contact surfaces
When to use which surface finish

Bacteria after cleaning Residues after cleaning

0.7 12
Bacteria (CFU/cm2)

Residues (µg/cm2)
0.6 10
0.5
8
0.4 Mechanically
6
0.3 polished surface
4
0.2

0.1 2

0 0
MP Ra<0.8 MP Ra<0.4 EP Ra<0.4 MP Ra<0.8 MP Ra<0.4 EP Ra<0.4

MP=Mechanically Polished
EP =Electro Polished Electro polished
surface

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Cleaning after polishing

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Temperature – Rouging - Bacteria

"While aquatic thermophiles capable of withstanding the hot water temperatures used
to sanitize pharmaceutical water system exist in nature, they do not exist in high
purity water systems because of the absence of their essential nutrients [98].
Designing sanitization cycles or monitoring techniques to kill these thermophiles, or
detect their presence, is considered unnecessary and an inefficient use of energy and
analytical resources."

ISPE Baseline Guide Water and Steam, Third edition, chapter 13, page 237. [Ref. 3]

José E. Martinez, "Hyperthermophilic Microorganisms and USP Hot water Systems",

Pharmaceutical Technology February 2004, pp.50-65 [Ref. 8]

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Temperature – ISPE [Ref. 3]
13.5.1.3 Heat sanitization with hot water
In high purity water, biofilm growth typically is either minimal or absent above approximately
45°C. Temperatures above approximately 50°C usually are hostile and slowly lethal. Higher
temperatures such as 65°C and 80°C often are used for hot water sanitization. EU GMP
(Annex 1, #59) (Reference 40, Appendix 1) suggests using greater than 70°C. The higher
the temperature, the quicker will be the microbial death……

….water distribution systems operating ≥ 65°C (149°F) generally are considered to be self-
sanitizing; 80°C (176°F) is an often-used target temperature.

Care should be exercised with 65°C sanitizations since heat losses can cause a
temperature drop of as much as 10°C to 15°C across a metal POU valve to downstream
moist surfaces, yielding minimally sanitizing conditions on those distal surfaces.

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Temperature – ISPE [Ref. 3]
13.5.1.4 Heat sanitization with steam
Steam can be used to sanitize water systems, although it is typically no more effective in
system microbial control than hot water and considerably more complicated. It increases
component wear, increases rouging in SS systems, and may have higher safety concerns
than hot water

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Bacteria classes
Growth rate vs temperature for five environmental classes of prokaryotes

Extreme Extreme
Moderate
thermophilic thermophilic
thermophile
bacterium archaeon

Mesophile
GROWTH RATE

Psychrophile

0 10 20 30 40 50 60 70 80 90 100 110 120

TEMPERATURE °C

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Bacteria and Temperature

Ex. of microorganisms that could exist in water systems:

Bacterium Optimum (°C)/Upper (°C)
Escherichia coli 37/45 Gram-negative
Staphylococcus aureus 30-37/45 Gram-positive
Pseudomonas maltophilia 35/41 Gram-negative
Pseudomonas aeruginosa 37/42 Gram-negative
Pseudomonas fluorescens 25-30/39 Gram-negative
Listeria monocytogenes 30-37/45 Gram-positive
Campylobacter jejuni 37-42/45 Gram-negative
Clostridium perfringens 37/50 Gram-positive
Shigella spp. 35-37/47 Gram-negative

Conclusion:
Microorganisms found in water systems can’t survive in temperatures > 50 °C

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Biofilm

• More than 99% of all microbial activity occurs in the biofilm

• The biofilm is a favorable habitat for bacteria as it allows
access to nutrients
• The complex biofilm matrix also works as a defense against
stress, e.g. temperature, shear forces, chemicals, etc.

Time

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Biofilm - Growth

Biofilm attach and grow more easily:

▪ When the surface roughness increase (Ra >0,8µm/30 μin)
▪ On hydrophobic, nonpolar surfaces (such as PTFE and other plastics)
▪ At low shear forces
▪ In crevices, pockets, and dead-legs
• Nutrition gets trapped
• Bacteria gets trapped

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Biofilm - Control

To minimize the biofilm:

▪ Minimise nutrition in the system
Hygienic design (right surface finish, no pocket, dead-legs or crevices)
▪ Secure high shear forces (velocity and hygienic design)
▪ Avoid hydrophobic nonpolar surfaces such as plastics

To remove the biofilm:

▪ Disinfectants in high concentration
▪ Drying the system completely

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Poll 4 & 5
4. If your facility have WFI distribution systems, which Temperature is it operated at?
▪ > 50°C
▪ > 65°C
▪ > 75°C
▪ > 85°C
▪ Don’t know

5. If your facility heat sanitize PW water systems, what is the sanitization temperature?
▪ < 75°C
▪ > 75°C
▪ > 85°C
▪ Steam
▪ Don’t know

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ASME BPE [Ref. 5]
NONMANDATORY APPENDIX D ROUGE AND STAINLESS STEEL

Considerations for reducing rouge formation

▪ System fabrication, Table D-2-1
▪ System operation, Table D-2-2

Evaluation methods to measure rouge

▪ Process fluid analysis, Table D-3.1-1
▪ Surface analysis, Table D-3.2-2

Methods to remediate the presence of rouge in a system

▪ Rouge remediation processes, Table D-4.1-1

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Reducing rouge formation - ASME BPE

2019 ASME BPE Table D-2-1 & D-2-2

Standard Copyright © 2019 by The
American Society of Mechanical Engineers

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Measure rouge - ASME BPE

2019 ASME BPE Table D-2-1 & D-2-2

Standard Copyright © 2019 by The
American Society of Mechanical Engineers

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Rouge remediation - ASME BPE

2019 ASME BPE Table D-2-1 & D-2-2

Standard Copyright © 2019 by The
American Society of Mechanical Engineers

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ASME BPE, De-rouging - Summary

Recirculation or soak or gel (40°C -80°C, 2-12 hours)

▪ Phosphoric acid (5%-25%)
▪ Citric acid (3%-10%)
▪ Phosphoric blends (5%-25%)
▪ Sodium hydrosulfite (-10%)
▪ Electrochemical cleaning (with phosphoric acid)
▪ (Class II rouge) Oxalic acid (2%-10%) - may etch the surface!

Followed by immediate deionized water rinse and passivation

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Poll 6
6. If you de-rouge your water system, which chemicals are you using?
▪ Phosphoric acid
▪ Citric acid
▪ Phosphoric blends
▪ Sodium hydrosulfite
▪ Electrochemical cleaning
▪ Others
▪ Don’t know

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References and further readings
Rouging
1. Mathiesen et al. Using exposure tests to examine rouging of stainless steel, Pharmaceutical Engineering,
June/August 2002. Vol. 21 No. 4
2. ISPE Critical Utilities D/A/CH COP. Rouge in pharmaceutical water and steam systems. Pharmaceutical
engineering, July/August 2009. Vol. 29 No. 4
ISPE
3. ISPE Baseline Guide Vol 4: Water & Steam Systems 3rd Edition, 2019
USP
4. NF Dec 2020 (USP-NF May 2021)
ASME BPE
5. ASME BPE-2019, Bioprocessing Equipment, The American Society of Mechanical Engineering, June 10, 2019
Bacteria & Biofilm
6. Anne M. Bronikowski et al. Evolution, 55(1), 2001, pp. 33 40
7. Frank Riedewald and Aidan W. Sexton, Biofilms Survival and growth of bacteria in compendial high purity water
systems: Pharmaceutical engineering Vol.27 No 1.
8. José E.Martínez, Hyperthermophilic Microorganisms and USP Hot Water Systems, Pharmaceutical Technology
February 2004

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Contacts
For questions or support

CH Lew , Sales Manager, KHI Engineering Pte Ltd

Mobile : +65 9831 4356
[email protected] | www.khiee.com

Maznah Ismawi, Sales Manager, Alfa Laval SIN/PH

Mobile: +65 9646 4934
[email protected]

Agus R. Adisuwondo, Regional Pharma Manager, Alfa Laval Indonesia

Mobile: +62 8111 3510301
[email protected]

Per-Åke Olsson, Global Industry Manager, Pharma & Biotech, Alfa Laval Corporate
Mobile +46 (0)722 171 226
[email protected]

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Poll 7 & 8
7. Would you be interested in another ISPE webinar from Alfa Laval?
▪ Yes
▪ No
▪ Maybe

8. Please check webinar topics that you might be interested in? (multiple answers)
▪ Microbial control in water systems
▪ Stainless steel and welding in pharmaceutical systems
▪ Hygienic system design
▪ Mixing and agitation in Biopharmaceututical
▪ Other
▪ None

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