TECHNOLOGY APPLICATION

Application of Total Organic Carbon Analysis to Cleaning Validation

K. M. JENKENS* , A. J. VANDERWIELEN +, J. A. ARMSTRONG,
L. M. LEONARD, G. P. MURPHY, and N. A. PIROS,

Quality Control, The Upjohn Company, Kalamazoo, Michigan

ABSTRACT: Cleaning Validation is the process of assuring that cleaning procedures effectively remove residue From manufacturing equipment /facilities below a predetermined level. This is necessary to assure the
quality of Future products using the equipment, to prevent cross- contamination and as a GMP requirement. Currently, Cleaning validation sample are measured using HPLC or spectrophtometric Method of analysis
which are Often time consuming and subject to a number of interference. Total Organic Carbon (TOC) analysis is a new Method which has previously only been applied to measurement of carbon residues on production
surfaces for Biopharmaceuticals. We have applied the Toc analysis method to a number of traditional pharmaceuticals Products including antibiotics, steroids, and antinaiseant in addition to biopharmaceuticals. The
method offers Extremely low detection capability (ppm and ppb), rapid sample analysis time and therefore quick turn- around of Production equipment and facilities. TOC analysis is also applicable to on- line analysis.
The method allows the Measurement of extraneous materials such as process intermediates, cleaning agents, and protein materials notPossible by other methods.

Introduction area such as nuclear power plant , semiconductor

Cleaning validation has been an area of increased
regulatory and industry scrutiny. This attention is war-
ranted given the increased use of multipurpose equip -
ment and an overall concern for quality of product. For
both industry and FDA inspectors the concern is the
same_______assurance that equipment is clean and that
product quality and safety are maintained. Cleaning
validation provides such assurance. One of the key
components of any cleaning validation strategy are the
analytical methods use measure Residuals. The
overall goal is to assure that the product does not
become adulterated from any source whatever ; interme-
diates , excipients and cleaning agents must therefore be
considered . In most cases , however, the active drug
ingredients are still the overriding consideration .
The measurement of total organic carbon (TOC)
content of contamination by organic compound is a new
method applied to cleaning validation . The method has
previously been used for analysis of high purity water in
__________________________
Received December 20, 1994. accepted for publication May 22, 1995
Presented at the PDA Annual meeting , Philadelphia, PA November
1994
Author to whom correspondence should be addressed: Western
Production and for pharmaceutical grade water for
injection .
In 1990 the use of TOC for pharmaceuticals in
regard to cleaning validation was first reported(1).
since that publication the use of TOCfor biopharmaceu-
ticals has been widely used. The same characteristics
that make TOC applicable for biopharmaceuticals are
Appropriate for traditoional pharmaceuticals .TOC has
low level detection(ppb), rapid analysis time, is low cost
as compared to other methods, and can detect all
carbon – based residuals.
Theory__ TOC
TOC analysis involves the oxidation of carbon and the
detection of the resulting carbon dioxide produced from
oxidation. This oxidation can be induced by a few
common method including photocatalytic oxidation (2),
chemical oxidation (peroxdisulfate or oxygen) (3) and
High temperature combustion (4) . The work presented
In this paper was accomplished by use of the shimadzu
TOC 5000 analyzer.
The Shimadzu TOC -5000 (Shimadzu instruments,
Columbia Maryland ) is an elemental analyzer capabel
of independent measurement of TC (Total Carbon) ,
TOC (Total Organic Carbon) and IC (Inorganic Car-
 Michigan University, Kalamazoo, MI 49008 -3842. bon ) in water. The Shimadzu instruments measures
* Author to whom correspondence should be addressed: Sterile Manu- organic specices by combustion to produce CO2. An
Fecturing , The Upjohn Company Kalamazoo, MI,49001. NDIR(non dispersive infrared )detector is then utilized
Chemistry Department , Western Michigan University, MI , 49008 to measure the resulting CO2 produce from combus-
Chemistry Department , Illonois University, Normal, IL, 61761. tion. This combustion takes place at high temperature
$ R. Baffi, et. Al.,1991 . A Total Organic Carbon Analysis for
Vidating Cleaning Between products in Biopharmaceutical Manu-
Fcturing Journal of Parenteral Science & Technology, Volume
45 (1), Pages 13-19

PDA Journal of Pharmaceutical Science & Technology

And produces CO2 by the reaction described below. Measurement of TC (Tital Carbon)

Reaction of Catalytic Combustion The TC combustion tube is filled with TC catalyst and
Heated to 68oC. Carrier (purified air) is supplied
(Oxidation of Organic Compound) into this tube after the flow rate is adjusted to 150
CaHbNcO + O2 - aCO2 __ +b/2H2O +cNO ml/ min by pressure and mass flow controllers and
Mistened by a humidifier.
When the sample is introduced in to the TC combus-
(Decomposition of Carbonate) tion tube via yhe sample injector (ASI- 5000), the TC
MeCO3 __ CO2 +MeO component in the sample (comprising TOC and IC ) is
(Decomposition of Bicarbonate) combusted /oxidized to from CO2.The carrier gas with
The combustion product (CO2 ) from the combustion
MeHCO __ CO2 + 1/2Me2O +1/2H2O flow through the IC reaction vassel and is cooled and
dried by a dehumidifier. It then passes through a
Sharp peak heights are obtained by instantaneously halogen scrubber into the sample cell of the non-
converting all carbon compound toCO2 as described. dispersive infrared detector ( NDIR ), where CO2 is
the TOC- 5000 uses paek area as the actual measure- detected.The NDIR outputs a detection signal ( analog
ment method. signal ) wich generates a peak, the area of wich is
Calculated by a data processor.
The peak is proportional to the TC concentration in
TOC/ TC/IC/ NPOC the sample . If a calibration curve expressing the rel-
The Shimadzu TOC- 5000 instrument(fig. 1) IS ca- ationship between peak area and the TC concentration
pable of performing total organic carbon analysis (TOC), is obtained using a TC standard solution, the TC con-
total carbon analysis ( TC ), inorganic carbon analysis entration of the sample may be determined. This is an
( IC) and non –purgable carbon analysis ( NPOC). External standard method of calibreation. The standard
each method is described further and are related by the curve used can be either polygonal or a liner least
following sample equation. Squares fit.

TC = TOC +IC
 NPOC = TOC + purgables

Figure1____ Schematic of the TOC 5000 instrument. The Schematic was reproductive from the instrument operating procedure with the
Permission of the Shimadzu corporation.

Vol. 50, NO. 1 / January __ February 1996

Measurement of IC ( Inorganic Carbon )
 TABLE I
Analysis Method ___ Common Uses
The sample is introduced via sample injector (ASI- 5000)Into the IC reaction
Cleaning
vessel ( containing IC reagent-
Drug Agent Biopharma-
Phosphoric acid ), through which carrier gas is following formtiny bubbles.
Method Residue Excipient Residue ceutical
Only the IC component in the
sample is the decomposed to from CO2 which is detected
upon reaching the NDIR. The concentration of IC is HPLC X X X
determined in the same manner as that of the TC except TLC X X
that an IC calibration curve is used to determine sample spectrophoto-
concenteration.Carbon in the form of carbonate and metric X X X
Hydrogen carbonate may be measured as TOC X X X X
ELISA X
electropho-
Resis X
PH X
Conductivity X
Measurement of TOC ( Total Organic Carbon) Gravimetric X X

The TOC concentration may be determined by sub-

Tracing the IC concentration , obtained as described
Above under measurement of IC ( Inorganic Carbon ) from
The TC concentration as describedunder measurement
Of TC ( Total Carbon ). This subtraction is performed by TABLE II
The TOC-5000 and reported as the toc value. Analysis Method _______ Advantages/ Disadvantages

Method Advantages Disadvantages

Measurement of NPOC ( non- purgable Organic Carbon
The TOC concentration may be determined directly HPLC highly specific Long analysis time
Using another procedure. In this case , the sample is moderate to high expensive
acidified beforehand and then spraged automatically sensitivity
with purified gas to remove the entire IC component. TLC Quantitative
The is then analyzed to obtained the TC concentro- Moderate to high visual endpoint
Tion as described under the section Measurement of TC Senstivity Long sample prep
(Total Carbon). This method is refrred to as the NPOC Fairly inexpensive
Method. Spectrophoto- Moderate to high Not quantitative
NPOC specifically refers to non- volatile organic car- metric specificity
bon which is not eliminated by ovaporation during the High sensitivity
sparging process. Volatile organic compound s such as method
organic solventwhich are not soluble in water TOC Broad spectrum Non- specific
are eliminated from a sample at room temperature via Low level detec- Aqueous soluble
sparging . The organic carbon which is evaporated during tion samples only
sparging is referred to as POC ( purgable Organic on- line capability
Carbon ). There is however an increase in analysis time Rapid sample
When performing NPOC on sample since the purge turn- around
Time with gas ( high purity air ) is 2 minutes or more minimal sample
Prep
ELISA specific for bio- Long sample turn
 Pharmaceutical around
Very sensitive very expensive
Labor intensive
Problems with
Advantages for cleaning Validation TOC analysis offers some distinct advantages for
Cleaining validation over other traditional method listed denatured pro-
In Table I. For each of these method s the common uses tein
Are listed in terms of residuals they detect. Of all the Electrophoresis Specific for bio- Very expensive
Methods TOC has the broadest range of uses since it can pharmaceutical Labor intensive
detect any carbon- based residuals. Table II list some of Moderate sensitive Problems with
the distinct advantages and disadvantages of the other denatured pro-
methods used for cleaning validation. The most signify- tein
can differences among the methods are in the areas of Long sample turn
cost and detection capabilities.HPLC has the capability around
for the lowest level of detection,but is also quite costly. PH Rapid Non- specific
Spectrometric methods ( UV ) and thin- layer chromatog Inexpensive Water soluble only
raphy (TLC ) are lower cost methods but at the expensive On- line capability Only useful for
of detection capability. Of the methods applied to Cleaning agents
biopharmaceutical, ELISA is a high cost, time- consum- Limited sensitivity
ing method. The electrophoresis method, polyacryl- Visual Detection Immediate result Not quantitave
amide gel electrophoresis (PAGE) is an extremely Good for genral Subjective
sensitive assay method. This method is primarily used Screening

PDA Journal of Pharmaceutical Science & Technology

Background
Material Description Particales ( PPM ) % Recovery

Quartz Wool+ Spun quartz Excessive 0.00 91.72

Absorbond TM * Hydroentagled poly- 30million particales/m2 0.00 29.01
ester
Cotton Prewashed in methanol High 9.71 66.29
And chloroform
Foam cubes* 5/ 16” foam cubes Low 61.9 __
Metrigard TM* Ulterafine glass fiber High 0.00 __
Acrylic binder
Clean cotton TM * sealed edge cotton 146 million particales/m2 5.50 48.83
Blend
Sterile Wipe HS II TM * Nonwoven cellulose/ 145 million particales/m2 7.42 74.43
Polyester blend
Allpha Wipe TM * 100% continuous fila- 30 millio particales /m2 0.0 72.89
Ment double knit
Polyester
Sterile WipeLP TM * 100% continuous fila- 30 million particales /m2 0.0 68.95
Ment polyester
Sterile Wipe LP 10 TM* 100 % continuous fila- 5 million particales/m2 0.0 82.21
 Ment polyester with
Thermal order seal
Sterile Wipe HS TM * Nonwoven cellulose and 165 million particales/m2 17.64 ___
Polypropylene com-
Posite
Beta Wipe TM* 70% polypropyl- 165 million particales/m2 10.78 ____
Ene / 30% cellulose

* Material obtained from the Texwipe Company; particale count is from Rexwipe literature.
** Material obtained from Gelman sciences.
+ Material obtained from Shimadzu through Giangarlo scientficCompany, Inc.

for the separation of protein based upon molecular
weight and can detect both variant and contaminating
proteins, yet identification is somewhat ambiguous . TOC
provides a method which is relatively inexpensive.rapid
in analysis time, and has low level detection ( ppm __ ppb ).
Since it detect all carbon based residues it can detect
Drug, excipient and cleaning agent residues. Given the
rapid analysis time (3__ 5 ) and the availability of
compact instrumentation it can be applicable to on- line
analysis.
There are some limitation of the TOC method. Only
Water soluble compounds can be analyzed by the method.
It is an Nonspecific method; however, that may not be a
Serious limitation since most of the result are negative
Due to the fact that the cleaning process is optimized and
The surfaces are clean. In these cases a specific method
Such as HPLC , TLC or any of the other specific assay
Methods are an unnecessary and added expense in terms
of assay time , cost and downtime. TOC provides an
immediate answer for validationand for on going moni-
torning at a lower cost. It provides assurance that the
surface of rinsate monitored is clean to a low residual
Level.
Assay development
Swab Challenge
As noted in the resent FDA guide to inspection of
cleaning process, “There are two genral types of
sampling that have been found acceptable. The most
desirable is the direct method of sampling the surface of
the equipment” (5 ). It was therefore desireable to de-
velop of swabbing technique for TOC analysis. The
swabbing technique involvesthe use of a swabbing
material, often saturatedwith a solvent, and physically
sampling the equipment or facility surface. In the case of

TABLE IV
Sample Fillter Materials __ Background
Millipore
Millipore Millipore Sterile Gelman Gelman Gelman
Millex LCR Millex- HV Millex- HA Acordisc GHP Acordisc
Filter 0.05um 0.45um 0.45um 0.45um Acirdisc CR

Composition Hydrophilic Durapore PVDF Cellulose PVDF Hydrpphilic PTFE

PTFE Polypropylene
Background 0.0* 3.2 0.0* 184.9 11.5 0.0
( ppm carbon )
% Recovery 106.9 101.1 100.2

• Background was noted below the detection limit of 1.4 ppm

TABLE V
Swab / Rinse Recovery

Drug Drug Swab % Recovery

Category Formulation Type/ indication Agent Swab/ Rinse

Biopharmaceuticals Sterile Recombinant protein Water 68 79

Sterile Immunosuppressant Tween 80/ Water 52 83
Antinauseant Dry product __ Tablet Motion sickness Water 83 103
Antibiotics Dry product __ Tablet Acidified Water 83 102
Sterile solution Anaeribic bacteria semi- Water 68 94
synthetic antibiotic
Dry product __ powder Broad spectrum antibiotic Water 84 103
Fluid
Sterile solution Gram- positive antibiotic Water 50 98
Dry product __ tablet
Steroids Sterile solution Steriodal research com- Acidified water 73 90
Pound
Sterile solution Glucocorticiod Water 68 89

TOC only water or agents with low carbon background
Can be used for swabbing. These agents can include
Acids, bases, low carbon surfactants and wetting agents
(Tween 80 ). In adition, theswab it self must be of such a
Material that it does not contribute significant back-
Ground. Both of these criteria present a challenge. A
Number of swabbing material were evaluated for use
With TOC. In all cases HPLC grade water was used as a
Solvent choice for the analysis. Table III lists material
Evaluated as possible swab, their composition, rated
Partical count information and the background level of
Carbon determined for two swab in 10.0ml of high
Purity water. In adition, the recovery of each swab was
Tasted using spiked sample of a water soluble drug
( methscopolamine bromide ). This evaluation demon-
Strated that quartz wool at the highest recovery with no
Detectable background. However, the quartz wool mate-
Rial left a residue of particulates on swabbed surfaces
Ans was therefore determined unacceptable.
The Alpha Wipe sterile Wipe, and sterile Wipe LP 10
Swab (The Texwipe Company, Upper saddle River,
NJ ) demonstrated the next best recovery with no signify-
Cant background. Since these swabs are a knitted polyes-
work. The alphawipe ( Texwipe Cat. No. TX 1004 ) was
used as the swab material since it was less costly than the
Sterile wipe and sterile wipe LP10which is the same
material only sterilized. The strile Wipe LP10 has a
manufacture-rated partical level less then the other two
Swab only due to the thermal border seal. This was not
significant since all swab material would be cut into
smaller size, thereby eliminating any advantage of a
sealed border. Alpha Wipe swab are prepared by cit-
ting sheet of the material into 2cm by 2cm square
which are then used as swab. NO pretreatment of the
material is necessary prior to use.
Sample Filtration
Since large insoluble particalesfrom swabbing surfaces
could be detected by TOC and cause mechanical prob-
lems in the sample injection system, sample filtration
Was necessary. Various filters from Millipore ( Millipore
Crop., Bedford, MA ) and Gelman ( Gelman Sci., Ann
Arbor, MI ) were tasted background. The results
( Table IV ) demonstrated that only the Millipore LCR,
Millipore HA, and Gelman CR filters had background
 Ter material there was no residue detected on the reading of 0.0 ppm carbon. For each of these filters
Swabbed area at the ppm level. This material was chosen studies were conducted to confirm that absorption of the
As the swabbing materialfor all further development drug did not occur on the filter material. The Millipore

TABLE VI
Swab / Rinse Recovery Over Range

Drug Drug Swab/ Rinse Swab / Rinse Swab / Rinse Swab/ Rinse
Category Type/ Indication Agent Slope Intercept Correlation
Coef.
Biopharmaceuticales Recombinant protein Water 0.884 1.004 __ 5.199 0.570 0.996 0.996
Immunosuppressant Tween 80/water 0.585 1.048 5.158 22.609 0.996 0.998
Antinauseant Motion sickness Water 0.758 1.119 3.882 5.138 0.999 0.999
Antibiotics Acidified water 0.834 1.031 3.454 _ 2.552 0.999 0.999
Anaerobic bacteria Water 0.668 0.988 _3.473 _6.705 0.999 0.999
Semisynthitic antibi-
Otic
Broad spectrum- antibi- Water 1.318 0.991 12.208 0.558 0.999 0.999
Otic
Gram- pissitive antibiotic Water 0.614 1.009 _ 0.569 _9.029 0.993 0.999
Steroids Steroidal research com- Acidified Water 0.765 0.981 3.122 _1.557 0.999 0.999
Pound
Glucocorticoid Water 0.769 0.962 _7.230 1.112 0.999 0.999

PDA Journal of pharmaceutical science & Technology

TABLE VII

Limit of
Drug Rinse Detection
Drug Category Type / Indication Agent (ppm range)

Biopharmaceuticals Recombinant protein Water 14

Immunosuppresant Tween 80/Water 6
Antinauseant Motion sickness Water 4
5
Antibiotics Anaerobic bacteria semisynthetic Water 8
Antibiotic
Broad spectrum-antibiotic Water 1
Gram positive-antibiotic Water 2
Steroids Steroidal research compound Acidified Water 7
Glucocorticoid Water 1
LCR and HA and Gelman CR filters all performed
Equally well in this regard . In the case of protein the
Gelman CR and GHP Acrodisc filers provided low
Background without absorption of the protein material.
Swab recovery
Six 1.0ml- sample of various drugat 225 mcg/ ml
Were spiked on stainless steel surfaces and allowed to
Dry. These sample were spiked onto an area approxi-
Mately225 cm2. swabbing was conducted using Alpha-
Wipes at the swab wetted with the slected swabbing
Agents. Two separate swab held with a stainless steel
Forceps were used to swab the 225 cm2 surface both
Vertically and horizontly.
The swab were placed in acid washed vails (Eagle
Picher, Miami, Oklahoma,) and 10 ml; of diluent (HPLC
Grade water ) was added. After extraction by shaking for
Ten minutes the solution was passed through a filter
( Millipore , Gilman, CR or GHP Acordisc )to remove
Any particulates. Swabsample were then analyzed by
The total carbon analysis method described. The average
Recovery from stainless steelsurfaces for the drugs
Tasted and the swabbing agent used are listed in Table V.
Swab recovery were greater than 50% in all cases,
Which was considered acceptable.
Determination of Swab and Rinse Recovery
The FDA guide to inspectionfor cleaning validation
Notes that “The firm should challenge the analytical
Method in combination with the sampling method ( S)
TABLE VIII
Comparison to HPLC
Drug
Drug Category Type/ Indication
Antinauseant Motion sickness
Antibiotics Anaerobic bacteria
Semisynthtic antibi-
Otic
used to show that contaminants can be recoverd from
the equipment surfaces and at what level, ie; 50 %
recovery 90%.” This reinforces importance of deter-
Mining the actual swab and rinse recovery of the
Method. These recovery valuesare then used in the final
calculation.
Rinse Recovery
Six 1.0ml sample of various drug at 225 mcg / ml
were spiked intostainless steel beaker and allowed to
dry. 20 ml of the solution used for swabbing was used to
rinse the beaker. This solution was collected and
filtered prior to analysis. Rinse recovery data for various
Drug compounds are listed in Table V. Although rinse
recovery values from 79__103%, recovery was
generally in the 90 % range of better.
These recovery values are considered a worst case
since the sample are allowed to dryon the surfaces. The
Swab and rinse recovery values are used in the final
Calculation for sample taken from production surfaces.
Linearity of detector response
Linearity of detector response was measured for
Potassium hydrogen phthalate standards diluted from a
1000 ppm stock solution. Potassium hydrogen phthalate
C8H5O4K ( Sigma, St. Louis, Mo. )provides a pure carbon
Source which was used a standard. The range of
detection measuredfor Linearity wasfrom 20 % to 400 %
of the 1.0mcg/cm2 level for each drug tasted in the ppm
range. The detector responsefor a typical analysis was
HPLC
TOC Average Average
Recovery RSD Recovery RSD
83.3 4.2 55 3.1
68 14 39 7.5
 Gram positive – antibiotic 50 9.9 86 6.4
Steroids Steroidal research com-
Pound
Glucocorticoid 71.6 8 72.4 7

TABLE IX
Comparison to UV Total protein methods
TOC UV Total
Drug Drug Average protien
Category Type Indication Surface Recovery RSD Average
Recovery RSD

Biopharmaceutical Immunosuppreseant Glass 51 19.5 21 32

Stainless steel 52 19 16 30
Recombinant protein Stainless steel 68 10.8 55 5.7

Linear over the range as determined by peak area
Response. The correlation for the detector response was
Always greater than 0.95 with an intercept that was not
Significantly different from zero. All standard curves are
Prepared using potassium hydrogen phthalate and
Sample are analyzed against these stored curves. Check
Solution over the range of testing are used to ensure
Accuracy of the curves.
Recovery Over Range
Recovery over the range of 20% to 400% of various
Drugs at 1.0 mcg/m2 was determined for both the
Swabbing and rinse method.For the swab method 1.0ml
Sample at 20% , 50%, 100%, 150%and 400% of the 1.0
Mcg/cm2 levelwere spiked onto 15cm by 15 cm areas on
Stainless steel sheet and allowed to dry. High purity
Water was used as a swabbing agent and Alpha Wipes
Were used as the swab. These result demonstrate that
the recovery was linear for all drugs tasted.
For the rinse method 2.0ml sampleat 20% , 50% ,
100%, 150%. And 400%of the 1.0mcg/cm2levelwere
Spiked into stainless steel beakers and allowed to dry.A
20.0ml rinse solution of HPLC grade water was added
To each beaker for recovery. The solution was agitated
And collected for analysis. The analysis of the rinse
Solution demonstrated that the recovery was linear
( Table VI ).
value of the intercept plus three times the estimated
standard daviation of the slop ( 6 ) . Table VII lists the
determined detection capability is only limited by the
tasted. These detection level were determined only for
the ppm range of the instrument. In the ppb range of the
instrument the detection capability is only limited by the
background of the process water used in that environ-
Ment . HPLC grade water used in the lab has a carbon
Background of 120__ 200ppb. The background of produc-
tion- purified rinse water will vary with the source and
Treatment . Analysis of production equipment rinses in
the ppb range should take account the background
carbon level of the water used for rinse.
Comparison to Other Methods
Comparison of the TOC method to existing HPLC
Methods for each drug was conducted to assure equiva-
Lency and controlled tested. Sample of drugs (1.0 ) ml
spiked onto 225cm2areas of stainless steel at a concen-
tration of225mcg/ml and allowed to dry.These areas
were swabbed and separate sample were analyzed by
both the HPLC and TOC method. Table VIII lists the
average recovery and RSD for six seprate sites ana-
lyzed by each method. In genral, the result were either
comparable or the TOC method had greater recovery
than the JPLC method.

Limit of Detection
Due to the background contributed by the Alpha Wipe
Swabs, the limit of detection for swabbing material. The swab
Ground contributed by the swabbing material. The swab
Background ranges typically from 0__15ppm . The swab
Background value from each run is used to subtract this
Background from all swab sample. Thereis no pretreat
Ment of the swab . The swab are handaled with powder –
Free class 100 clean rooms latex gloves (QRP, Tucson,
AZ ) and acid washed stainless steel foreceps. The gloves
Are powder – free , have low particulates and thereby do
Not contribute to the background for the swabbing
Agent.
In the case of proteins the TOC method was comp-
Pared to spectrophotometric total proteins method.
These total proteins methods both used the lowery method
for total proteins determination. The lowry method
suffers from interference due to surfactants and other
organic compounds (7 ). Table IX list the everage recover-
ery and RSD for each method. In all cases the TOC
method resulted in higher average recovery. One signify-
TABLE X
Production Monotoring Environmental Background
_________________________________________________
Results
Surface area ppm/225 cm2
_________________________________________________

To determine the rinse limit of detection , solution Sterile filling pump 0

Were prepared at 10% , 70% and 200% of the 100% level Sterile holding tank 2
( 225 mcg drug ), spiked into 1.2 liter stainless steel Sterile trays 3.8
Beakers and allowed to dry. The beaker were rinsed Blender interior 0
With 10.0 ml rinse solution (water ) and analyzed for Blender lid 4
Total carbon. The results were graphed and the intercept Loading chute 0
Calculated. The detection limit is defined as the absolute Blending Rm Floor 60
Module Floor 177
___________________________________________________

PDA Journal of pharmaceutical science& Technology

TABLE XI TABLE XIII

Freez Dryer Trays Sterile Manufecturing Tank __ protein
_______________________________________________________ __________________________________________________
Equipment Antibiotic Steroid Equipment Run 1 Run 2
Part Result (ppm ) Result ( ppm) Part Result (ppm) Result (ppm)
_______________________________________________________ __________________________________________________
Tray 1 interior 2 9 Tank interior 4 10
Tray 1 lid 5 3 Tank lid <1 4
Tray 2 interior <1 3 Tank discharge port <1 6
Tray 2 lid <1 <1 Qs rod 13 7
Tray 3 interior 3 <1 ___________________________________________________
Tray 3 lid 1 4
________________________________________________________
Cant advantage to the TOC method is that it detect both
Native and denatured protein.
Environmental Background
To determined areas where TOC analysis was feasible
It was necessary to determined the background for produc-
Tion surfaces. Background sample were taken from
Equipment in the sterile operation and dry products
Manufacturing area. In the sterile area swb sample
Were taken from filling pumps, holding tanks, and sterile
Trays ( Table X ). In the dry products area, samples were
Taken from the interior of blenders, the lid and the loading
Chute. In both area sample were taken from the
Module and facility floor . The equipment surfaces all
Had background carbon levels from 0__4 ppm over a 225
Cm2 area. Floor surfaces were in the range from 60__ 177
Ppm of carbon over a 225 cm2 area. This data demon-
strates that TOC would not be optimal for monitoring
non product contract surfaces such as the facility; how-
ever, it is appropriate for the equipment. This was not
viewed as a serious disadvantage since facilities can be
visually inspected and are usually only required to meet
less stringent resodue limit.
Production Sampling
The TOC method developed were used for cleaning
Validation monitorings in various areas of production. In
The sterile operation the TOC analysis for antibiotics
Steroids and proteins were used to monitor sterile
Operation equipment such as bulk freez dryer trays ,
Sterile manufacturing pumps and sterile manufacturing
Tanks. Tables XI, XII, XIII, XIV, and XV list data for
Each of these equipment surfaces. In all cases a 225 cm2
Area was swabbed using Alpha Wipe and the swab were
Tested by TOC analysis. In all cases the equipment
Monitored met test in regard to residual monitoring
Limits.
TABLE XII
Sterile Manufecturing Pump___ Protien
_____________________________________________________
Equipment Part Result ( ppm )
_____________________________________________________
In the fluid manufacturing area the TOC analysis
Method for an antibiotic was used to validate a new
cleaning process. Table XIV shows the data from this
Analysis which indicates high result for the tank interior
And impeller from two separate monitoring. This data
of the shaft which was identified as a cleaning problem
TOC analysis has also been applied in the dry prod-
ucts area. Table XV lists the monitoring result for a
feeder used in one of our micrinizing operations. The
result were all 10ppm or less for each 225 cm2 area
monitored by swabbing except for one sample site
These result demonstrate that TOC analysis of swab
samples can be effectively used for cleaning validation of
equipment in the sterile, fluid,/ ointments and dry prod-
ucts manufacturing/ production area.
Determinetion Cleaning Agent
The FDA guide to inspection to cleaning validation
notes the importance of determining residual cleaning
agents with their statement “ As with products residues, it
is imported and it is expected that the manufacturer
Evaluate the efficiency of cleaning process for the
the removel of residues. Detergents are not part of the
manufacturing process and are only added to facilities
cleaning during the cleaning process” (5). Because of
these concern residual cleaning agents should be exam-
ined in the cleaning process. TOC is one possible
analytical method to detect residual cleaning agents
since they are carbon based.
The carbon content of cleaning agent was deter-
mined by making up 100 ppm solution of each of the
cleaning agents thought to contain carbon , and determin-
ing their carbon content using the shimadzu TOC-5000.
The TC content of the samples were measured and used
the determine thr present carbon in each of the cleaning
agents tested. The TOC analysis works best under acidic
Condition due to the platinum catalyst in the combus-
Tion. In circomastances where the sample run are
TABLE XIV
Fluid Manufacturing Tank ___ Antibiotic
________________________________________________
Equipment Run 1 Run 2
Part Results ( ppm ) Results (ppm)
Filter housing 9 ________________________________________________
Filter housing top 3 Tank interior 7 82
s.s. pipes 3 Impellor 66 9
Rotors 9 Tank discharge port 15 10
Rotor housing 6 ________________________________________________
______________________________________________________

Vol. 50, No. 1 / January_ February 1996

TABLE VX CIP/COP have lead to further use in the solid dose,

Micronizing Feeder ___ Antibiotic fluid / ointments and bulk drug operation.
_______________________________________________________ TOC has the capability for no-line testing of such
Equipment Run 1 Run 2 system. This would provide advantages such as rapid
Part Results (ppm ) Result (ppm ) response in term of resultand provides a means to
_______________________________________________________ troubleshoot the cleaning process. A stainless steel
Mixing vessel was used to simulate a manufacturing
Feeder top 0 1 vessel. High purity water recirculated through the vassel
Hopper 6 1 was tested to residual carbon using a CSM-5000 continu-
Lower feed chamber 9 0 ous sampling module attached to the Shimadzu TOC-
Feed screw 2 0 5000. After a period of time during which the recycled
Feed screw housing 16 0 water in the vessel was tested, a standard carbon
Blade 5 0 solution was spiked into the vessel . This tested was
________________________________________________________ performed by NPOC analysis. The use of on-line TOC
Could be for no-line quality control, cleaning process
Neutral or slightly acidic injection of 0.2 N HCL are run auditing validation, and/ or troubleshooting the clean
Approximately every 20 samples, but since many of the ing process during method development, on-line TOC
Cleaning agents have PHs above 7, Hcl injection were would allow eximanition of the point at which the bulk
Run more often. The results of this study, including the of the residual is removed by the cleaning process
Ppm carbon determined and percentage carbon for each thereby providing optimization of the cleaning cycle in
Prepared solution, can be seen in table XVI. The major terms of wash and rinse time.
Components of each cleaning agents are listed for each A study of different cleaning agents rinse cycle was
Trade name cleaning agent. Most of the cleaning agents conducted using this on-line approach. For three differ-
Tested could be detected at the 2_20 ppm carbon range ent rinse cycle the total carbon, chloride ion and
For a 100 ppm solution. The only exception was Delvak, phosphate level was measured by TOC and ion Chroma
Krystall and sentol which could only be detected at tography in the rinse water from the process. This data
Solution concentration of 1000 ppm. Those cleaning ( Table XVII ) showed that two short cycle were more
Agents with low carbon percentage may not be suitable effective thanone longer rinse cycle of removing both
Candidates for detection by TOC. Other methods such drug and cleaning agents. This type of testing allows the
As ion chromatography may be more appropriate in such ability to actually study the effectiveness of the cleaning
Cases. Process and thereby optimize the process.

On-line Capability
CIP (cleaning in place) and COP (clean out of place)
System were first introduced in the dairy industry.
These system are used more frequently in our industry.
They provide a consistent level of cleaning. A few years
Ago they were primarily used in the parentral drug
Areas. However, recent advances and decreased costs in
Conclusions
Total Organic Carbon analysis provides a new method
for cleaning validation. The methods offer s low level
detection, rapid analysis time, and detects all carbon

TABLE XVI
Cleaning Agent Carbon Content
Cleaning Concentration Cleaning Agent (ppm) Percent
Agent (ppm) Composition Measured Carbon
Airkem F/ H 98.80 Quaternary ammonia 7.420 7.510
Adept 105 Potassium hydroxide 4.363 4.16
Alconox 100.2 Phosphates sodium carbonates 12.66 12.64
sulphonates
CIP 100 109 Potassium hydroxide 4.809 4.41
Delvak 962 Sodium hydroxide tripolyphos- 14.37 1.49
phate sodium carbonate
Grease Ease 104.8 Sodium metasilicate butyl cel- 11.45 10.92
losolve
Krystall (immiscible in water) 925 Oil base 1.229 LT 1
Resource 102 Sodium hydroxide 2.976 2.92
SD-20 114 Sodium phosphate sulfonates 4.984 4.37
etyhoxylated amide
Sentol 1135 Phosphoric acid 4.625 LT 1
Spray Nine 109 Sodium metasilicates 4.704 4.32
SDT 99.60 Sodium dichlorotriazene 15.71 15.77
trione
Sprex 102.3 Sodium carbonate sodium sili- 10.48 10.24
cates
TBQ 104.0 Alkylamine ammonium chlo- 19.32 18.58
ride

PDA journal of pharmaceutical Science & Technology

TABLE XVII References

Cleaning Process ___ Rinse Study
____________________________________________________________ 1. R. Baffi, et al; “ A total organic carbon analysis method for validat-
Rinse Duration TOC CI Ion Phosphate ing cleaning between products in biopharmaceutical manufectur-
 Cycle (minutes) (ppm) (ppb) (ppm) ing, “Journal of parentralScience and Technology,45 (1), 13 _19 ___________________________________________________________
(1991).
Run 1 4 50 994 6.7 2. R.W. Mathews, M. Abdullah, and G. K.- C.Low, “ Photacatalttic
Run 2 (cycle 1) 2 16 698 2.6 oxidation for total organic carbon analysis,” Analytica Chimica Acta
Run 2(cycle 2) 2 6 220 1.4 233, 171__179 (1990).
_________________________________________________________________________ 3. ASTM Method D4 129__82, “ Determination of dissolved organic
Carbon,” ASTM book of standard, Philadelphia, Pa. (1984)
based residual at a moderate costs. The method provide 4. Y. sugimura, Y. Suzuki, “ A high temperature catalytic oxidation
Linear detection over the range studied and acceptable method for the determination of non-volatile dissolved organic
recovery for various pharmaceutical tested. Data from carbon in seawater by direct injection of a liquid sample,” Marine actual production monitoring demonstrate that TOC
Chemistry 24, 105__131 (1988).
can be used effectively for cleaning validation, In short 5. FDA, “ Guide to inspection of cleaning validation process
TOC method can be developed, validated and utilized (Division of Field investigation, office of regional operarions,
for pharmaceutical cleaning validation. Office of rtegulatory affairs, july 1993).
6. J. K. Tylor, “ Quality assurance of chemical measurements,”
Lewis publisher, Chelsea, MI, p. 78__84, 1987
7. Lowry, et al; Journal of bio.Chem; 1951, (193) , p. 265__ 275