Pharmaceutical Related

7.7 USP-Specified TOC System Suitability Test (1) - TOC

■Explanation ■TOC System Suitability Test Data by

The United States Pharmacopeia (USP) specifies the use of USP Method
Total Organic Carbon (TOC) for management of organic The TOC system suitability test was conducted using
impurities in purified water (PW) and water for injection the Shimadzu TOC-LCPH combustion catalytic oxidation
(WFI). According to the USP, the TOC analyzer to be used type analyzer by the procedure outlined in Table 7.7.1.
for these analyses must satisfy the TOC system suitability The instrument was calibrated beforehand using aqueous
testing requirement, and must be capable of detecting solutions of potassium hydrogen phthalate with carbon
TOC at concentrations below 0.05 mg/L.Here, using the concentrations of 0 and 0.5 mgC/L, respectively. The
Shimadzu TOC-LCPH combustion catalytic oxidation type TOC system suitability test data are shown in Fig. 7.7.1.
analyzer, we introduce examples of TOC system suitability According to the USP, the detection rate is to be evaluated
testing and measurement of TOC at a concentration below using the analyzer response values, but here, the measured
0.05 mg/L. concentrations were used instead.
The result indicated a 100.1 % detection rate with respect
■TOC System Suitability Test Specified in USP to the system suitability test solution (1,4-benzoquinone
The TOC system suitability test indicated in the USP aqueous solution), thereby satisfying the system suitability
specifies the use of two types of USP reference standards test requirement. (Table 7.7.2)
(sucrose and 1,4-benzoquinone). Sucrose is used as a test
solution standard, and 1,4-benzoquinone is used as the ■Analytical Conditions
system suitability test solution.In addition, calibration Instrument : Shimadzu TOC-L CPH Combustion
of the TOC analyzer is specified to be conducted using Catalytic Oxidation Typse Analyzer
a method that is suitable for that instrument. The test Catalyst : High-sensitivity catalyst
procedure is shown in Table 7.7.1.
Injection Volume : 816 μL
Measurement Item : TOC (=NPOC: TOC by acidification/
Table 7.7.1 TOC System Suitability Test Procedure Specified in USP
sparging)
TOC system suitability test procedure Calibration Curve : 2-point calibration curve using 0-
(1) Measure the TOC in distilled water (distilled water 0.5 mgC/L potassium hydrogen
used for preparing test solution), This value is phthalate aqueous solution
indicated as rw.
(2) Measure the TOC in the sucrose standard solution
(0.50 mg/L carbon concentration). This value is
indicated as rs.
(3) Measure the TOC by the system suitability test
(1,4-benzoquinone solution with 0.50 mg/L carbon
concentration). This value is indicated as rss.
(4) The system suitability test requirement is satisfied if:
detection rate = 100 (rss - rw) / (rs - rw) is 85 % - 115 %

Fig. 7.7.1 TOC System Suitability Test Data

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7.7 USP-Specified TOC System Suitability Test (2) - TOC

Table 7.7.2 Results of TOC System Suitability Test ■Analytical Conditions

(1) Distilled water TOC value rw = 0.0205 mg/L Instrument : Shimadzu TOC-L CPH Combustion
(2) Sucrose standard solution TOC value rs = 0.5173 mg/L Catalytic Oxidation Type Analyzer
(3) System suitability test solution (1,4-benzoquinone Catalyst : High-sensitivity catalyst
aqueous solution) TOC value rss = 0.5176 mg/L Injection Volume : 816 μL
(4) System suitability test solution detection rate Measurement Item : TOC (= NPOC: TOC by acidification/
= 100 (rss - rw) / (rs - rw) sparging)
= 100 (0.5176 - 0.0205) / (0.5173 - 0.0205) Calibration Curve : 2-point calibration curve using
= 100.1 % 0-0.5 mgC/L potassium hydrogen
phthalate aqueous solution
Sample : 0.025 mgC/L potassium hydrogen
■TOC Measurement Below 0.05 mg/L phthalate aqueous solution
The USP specifies that TOC analyzers to be used must be
able to detect TOC at a concentration below 0.05 mg/L.
Table 7.7.3 Measurement Data for TOC Concentrations Below
To verify this, we measured a potassium hydrogen 0.05 mg/L (50 μg/L)
phthalate aqueous solution with a TOC concentration of
0.025 mgC/L. The results are shown in Fig. 7.7.2 and TOC Value Coefficient of
Sample Name
[mgC/L] Variation (CV) [%]
Table 7.7.3. Because the distilled water used to prepare
the sample contained TOC components as impurities, Potassium hydrogen
the measurement resulted in a higher concentration of phthalate aqueous 0.047 2.66
solution
0.047 mgC/L, with a resulting coefficient of variation
(CV) of 2.66 %. Since the coefficient was within 10 %, a
CV value commonly associated with a concentration close
to the lower limit of quantitation, the Shimadzu TOC-
LCPH combustion catalytic oxidation type analyzer clearly
satisfied the USP requirement for TOC measurement of
concentrations below 0.05 mg/L.

Fig. 7.7.2 Measurement Data for TOC Concentrations Below

0.05 mg/L (50 μg/L)

98
 Pharmaceutical Related

7.8 Cleaning Validation by TOC Analyzer (1) - TOC

■Explanation ■Preparation of Residue Measurement Sample

To ensure quality control and safety in manufacturing In order to evaluate the cleaning validation sampling
facilities within the pharmaceutical industry, it is methods, residue measurement samples were created by
important that cleaning validation be conducted following applying various types of pharmaceutical products and
the cleaning of production-related equipment. Cleaning their constituents to stainless steel pots. The aqueous
validation ensures that the quantity of residual substances and non-aqueous substances that were used are listed in
collected from the surfaces of the equipment is within Table 7.8.1. The aqueous substances and non-aqueous
permissible limit. Depending on the sampling method and substances were dissolved in water and ethanol or acetone,
measurement method used for this cleaning validation respectively, and the solution concentrations were adjusted
using a TOC analyzer, the following 3 types of methods to 2000 mgC/L (= carbon concentration of 2000 mg/L).
are available. 100 μL of each solution was then applied to a 5 cm by 5
(1) Rinse sampling–TOC measurement method cm squares area on the surface of a stainless steel pot, and
(2) S w a b s a m p l i n g – a q u e o u s e x t r a c t i o n – T O C the respective solvents were dried out to produce residue
measurement method measurement samples. Thus, the amount of carbon in
(3) S w a b s a m p l i n g – d i r e c t c o m b u s t i o n c a r b o n the sample at each application site was 200 μg. Among
measurement method these, Gentacin ointment (aminoglycoside antibiotic) and
Here we introduce the features of each of these methods, Rinderon ointment (corticosteroid) were prepared based
using the TOC-LCPH total organic carbon analyzer in the on determination of their carbon concentrations using the
measurement of residual pharmaceutical products and Shimadzu total organic analyzer system including the solid
their constituent substances. sample combustion unit.

Table 7.8.1 Sample Types

Substance Name Solubility in Water Solvent Used in Solution Preparation
Tranexamic acid Soluble Water
Anhydrous caffeine Soluble Water
Isopropylantipyrine Insoluble Ethanol
Nifedipine Insoluble Acetone
Gentacin ointment Insoluble Ethanol
Rinderon ointment Insoluble Acetone

■(1) Rinse Sampling–TOC Measurement Method

The Rinse Sampling – TOC Measurement method is a The results are shown in Table 7.8.2. Water-soluble tranexamic
technique in which the final rinse water used in the cleaning of acid and anhydrous caffeine had high recovery rates as
a production equipment unit is used as the TOC measurement expected. Moreover, water-insoluble isopropylantipyrine and
sample. This method is suitable for systems that cannot easily nifedipine had high recovery rates. However, recovery rates
be disassembled, such as CIP (clean-in-place) equipment of Gentacin ointment and Rinderon ointment were both low,
and narrow tubing. However, sampling is considered to be at less than 20 %. From these results, it is clear that evaluation
difficult if the residues are not soluble in water. To evaluate of the rinse water using this method is unreliable due to the
the recovery of the various substances when using the Rinse variation of recovery of substances which are not readily
Sampling – TOC Measurement method, 100 mL of pure water soluble in water.
was transferred to the stainless steel pot with the patch of
dried sample, and after stirring with a stirrer for 15 minutes to
prepare the rinse solution, TOC measurement was conducted.
■Analytical Conditions
Some of the measurement data are shown in Fig. 7.8.1. Instrument : S h i m a d z u T O C - L C P H To t a l
Since the carbon content in each of the residue measurement Organic Carbon Analyzer
samples is 200 μg, the TOC concentration would be 2 mgC/ Catalyst : High sensitivity catalyst
L if all of the sample were to dissolve in the water. Now, for Measurement Item : TOC (=TOC by acidification
the blank, measurement was conducted in the same way using sparge processing)
water that was transferred to the stainless steel pot, which in Calibration Curve : 2-point calibration curve using
this case had no patch of dried sample applied to its surface. 0-3 mgC/L potassium hydrogen
The measured blank concentration was subtracted from each phthalate aqueous solution
TOC concentration, and then compared to the theoretical value Injection Volume : 500 μL
of 2 mgC/L to determine the rate of recovery.
99
7.8 Cleaning Validation by TOC Analyzer (2) - TOC

Table 7.8.2 Measurement Results for Rinse Sampling – TOC Measurement Method
TOC Concentration Recovery Rate,
Substance Name
[mgC/L] [TOC Conc. – Blank / Theoretical Conc.]
Blank 0.030 -
Tranexamic acid 2.14 105 %
Anhydrous caffeine 2.19 108 %
Isopropylantipyrine 2.20 109 %
Nifedipine 2.17 107 %
Gentacin ointment 0.117 4.35 %
Rinderon ointment 0.333 15.2 %

Tranexamic Acid Isopropylantipyrine Gentacin Ointment

Fig. 7.8. Measurement Data Using Rinse Sampling – TOC Measurement Method

■(2)Swab Sampling–Water Extraction–TOC Measurement Method

The Swab Sampling –Water Extraction–TOC Measurement way by wiping the stainless pot which had no sample
method, as illustrated in Fig. 7.8.2, consists of wiping the applied before conducting extraction. The measured blank
inside surface of the production apparatus with a fibrous concentration was subtracted from each TOC concentration,
swab material, extracting the adhering material with water, and then compared to the theoretical value of 2 mgC/L to
and conducting TOC measurement of the extract solution. determine the rate of recovery. The results are shown in
Since the residue is physically wiped off from a fixed area Table 7.8.3. Water-soluble tranexamic acid and anhydrous
of the surface of the apparatus using the swab material, and caffeine had high recovery rates as expected. Moreover,
then analyzed, the sampling efficiency is high. However, water-insoluble isopropylantipyrine and nifedipine had high
because water is used for extraction of the residue, residues recovery rates of about 90 %. However, recovery rates of
that are insoluble in water are difficult to extract. Accordingly, Gentacin ointment and Rinderon ointment were both low, at
cleaning evaluation with respect to these residues may be less than 10 %. From these results, it is clear that evaluation
difficult for the same reason as that described with respect to of the rinse water using this method is unreliable due to the
difficultto-dissolve substances in the (1) Rinse Sampling – variation of recovery of substances which are not readily
TOC Measurement method. To evaluate the recovery of the soluble in water.
various substances when using the Swab Sampling – Water
Extraction –TOC Measurement method, the sample, which
was applied to a stainless steel pot, was wiped off with a 5 cm
■Analytical Conditions
by 5 cm squares piece of fibrous swab material, which was Instrument : Shimadzu TOC-LCPH Combustion
then placed in a glass jar containing 100 mL of pure water. Total Organic Carbon Analyzer
The residue was then extracted by stirring with a stirrer for Catalyst : High sensitivity catalyst
1 hour, after which TOC measuremet was conducted. Some Measurement Item : TOC (=TOC by acidification
of the measurement data are shown in Fig. 7.8.3. Since the sparge processing)
fibrous swab material (Alpha 10 obtained from Texwipe Co.) Calibration Curve : 2-point calibration curve using
that was used is made of polyester, very little organic material 0-3 mgC/L potassium hydrogen
is extracted from the swab itself. Since the carbon content in phthalate aqueous solution
each of the residue measurement samples is 200 μg , the TOC Injection Volume : 500 μL
concentration in the extraction solution would be 2 mgC/L if
Swab Material : 5 cm by 5 cm squares piece of
all of the sample were wiped off. Texwipe Alpha 10 swab material
For the blank, measurement was conducted in the same washed in pure water and dried
100
 Pharmaceutical Related

7.8 Cleaning Validation by TOC Analyzer (3) - TOC

Swab Sampling Water Extraction Volume Adjustment TOC Measurement

Fig. 7.8.2 Swab Sampling–Water Extraction–TOC Measurement Method

Table 7.8.3 Measurement Results for Swab Sampling–Water Extraction–TOC Measurement Method
TOC Concentration Recovery Rate,
Substance Name
[mgC/L] [TOC Conc. – Blank / Theoretical Conc.]
Blank 0.059 -
Tranexamic acid 2.19 107 %
Anhydrous caffeine 2.23 109 %
Isopropylantipyrine 1.90 92.2 %
Nifedipine 1.86 89.9 %
Gentacin ointment 0.093 1.70 %
Rinderon ointment 0.208 7.45 %

Tranexamic Acid Isopropylantipyrine Gentacin Ointment

Fig. 7.8.3 Measurement Data Using Swab Sampling–Water Extraction–TOC Measurement Method
■(3)Swab Sampling–Direct Combustion Method
The Swab Sampling – Direct Combustion method, as illustrated in wiping the stainless pot which had no sample applied. The measured
Fig. 7.8.4, consists of wiping the inside surface of the production blank value was subtracted from each TC value, and then compared
apparatus with a piece of inorganic quartz glass filter paper to the theoretical value of 200 μg to determine the rate of recovery.
swab material, and then conducting measurement using a direct The results are shown in Table 7.8.4. A high recovery rate of about
combustion carbon measurement system. The swab material with the 100 % was obtained for all the substances, regardless of whether they
adhering residue is merely placed in the sample boat, and the carbon were watersoluble or water-insoluble.
content is measured directly by the TOC analyzer with the connected
SSM-5000A Solid Sample Combustion Unit. By using this method, ■Analytical Conditions
water-insoluble residues that are difficult to extract in water can also Instrument : Shimadzu TOC-LCPH Total Organic Carbon
be collected, and measurement can be quickly and easily conducted Analyzer + SSM-5000A Solid Sample
without the need for any pretreatment, such as sample extraction, Combustion Unit (IC circuit bypass using
etc. To evaluate the recovery rate of the different types of substances system with cell switching valve set)
using the Swab Sampling –Direct Combustion method, we used Cell Length : Short cell
the quartz glass filter paper swab material to wipe off the sample Ssm Carrier Gas : 400 mL/min oxygen gas
adhering to the stainless steel pot, placed the swab in the SSM- Measurement Item : TC
5000A sample boat, and conducted TC measurement. Some of the Calibration Curve : 1-point calibration curve using 30 μL
measurement data are shown in Fig. 7.8.5. Since the carbon content of 1 % C glucose aqueous solution
in each of the residue measurement samples is 200 μg, the TC value Swab Material : Advantec QR-100 quartz glass
would be 200 μg if all of the sample were wiped off. filter paper (diameter 45 mm) heat-
101 For the blank, measurement was conducted in the same way by
treated at 600 °C for 15 minutes
7.8 Cleaning Validation by TOC Analyzer (4) - TOC

Quarts glass filter paper

Swab Sampling Transfer to sample boat Carbon measurement using SSM-5000A

Fig. 7.8.4 Swab Sampling –Direct Combustion Method

Table 7.8.4 Measurement Results for Swab Sampling –Direct Combustion Method
Recovery Rate,
Substance Name TOC Value [ C]
[TC Value – Blank/Theoretical Value]
Blank 0.00 -
Tranexamic acid 202 101 %
Anhydrous caffeine 201 100 %
Isopropylantipyrine 210 105 %
Nifedipine 212 106 %
Gentacin ointment 200 100 %
Rinderon ointment 209 104 %

Tranexamic Acid Isopropylantipyrine Gentacin Ointment

Fig. 7.8.5 Measurement Data Using Swab Sampling–Direct Combustion Method

■Conclusion
The measurement methods used here and their respective evaluation using these methods would be difficult for
recovery rates are summarized in Table 7.8.5. When using substances with low recovery rates. In contrast to that,
the Rinse Sampling–TOC Measurement and the Swab high recovery rates were obtained for all the substances
Sampling–Water Extraction–TOC Measurement method, when using the Swab Sampling–Direct Combustion
substances that do not easily dissolve in water were found method, regardless of whether the substances were water-
to include those that had high recovery rates, and those soluble or water-insoluble, thereby permitting residue
that had low recovery rates. This may be due to differences evaluation. Therefore, this method is considered to be an
in the strength with which the substances adhere to the effective measurement method for conducting cleaning
stainless steel pot. Accordingly, it is probable that residue validation.

Table 7.8.5 Summary of Measurement Results

Recovery Rate
Substance Name Solubility in Water Swab Sampling –
Rinse Sampling – TOC Swab Sampling – Direct
WaterExtraction – TOC
Measurement Method Combustion Method
Measurement Method
Tranexamic acid Soluble 105 % 107 % 101 %
Anhydrous caffeine Soluble 108 % 109 % 100 %
Isopropylantipyrine Insoluble 109 % 92.2 % 105 %
Nifedipine Insoluble 107 % 89.9 % 106 %
Gentacin ointment Insoluble 4.35% 1.70 % 100 %
Rinderon ointment Insoluble 15.2% 7.45 % 104 %
102