NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

Mitomycin C on Surfaces 9212

C15H1 8N4O5 MW: 334.33 CAS: 50-07-7 

METHOD: 9212, Issue 1 EVALUATION: FULL Issue 1: 30 May 2020

OSHA: None established

NIOSH REL: None established

PROPERTIES: Solid; purple colored crystal; MP >360 oC [1]

SYNONYMS: (not all inclusive) Mutamycin; Ametycine; [6-Amino-8a-methoxy-5-methyl-4,7-dioxo-1,1a,2,4,7,8,8a,8boctahydroazireno[2',3':3,4]pyrrolo[1,2-a]indol-8-yl]methyl carbamate.

SAMPLING

SAMPLER: Polyester swab or filter paper wipes

SUFACE AREA: 100 cm2

PRESERVATIVE: None

SHIPMENT: Ship frozen using dry ice

SAMPLE

STABILITY: >8 days (see Evaluation Section for

discussion)

FIELD BLANKS: 2 to 10 per set

ACCURACY

RANGE 

STUDIED: 0.1 to 25 μg/100 cm2 (See Tables 1 and 2).

BIAS: None established.

OVERALL

PRECISION (SS�rrrr): See Tables 1 and 2.

ACCURACY: Full recovery (90 to 107%) was established 

by spiked matrix-matched solutions at up 

to 6 different concentration levels and 

using 3 surfaces (See Tables 1 and 2). The 

precision by relative standard deviation 

was no larger than 7.5% at any 

concentration level (See Tables 1 and 2).

MEASUREMENT

TECHNIQUE: HIGH-PERFORMANCE LIQUID 

CHROMATOGRAPHY - TANDEM MASS 

SPECTROMETRY

ANALYTE: Mitomycin C

HPLC - COLUMN: C18 (150 x 3 mm, 3.5 μm) equipped with same 

type guard column (4 x 2 mm) and 2-micron 

precolumn filter

MOBILE PHASES: A = 10/90/0.1% (v/v/v)

acetonitrile/water/acetic acid

B = 75/25/0.1% (v/v/v) acetonitrile/water/acetic 

acid

FLOW RATE: 0.4 mL/min (0.5 mL/min post run)

GRADIENT: Time (min) Mobile Phase Composition

 0 to 12 0 to 70% B 

 12 to 13 70 to 100% B

 3 to 16 100% B Flow increased to 0.5 mL/min

 16 to 18 100 to 0% B (re-equilibration, 0.4 

mL/min)

 18 to 24 0% B (re-equilibration)

INJECTION VOLUME: 5 uL

IONIZATION SOURCE: Electrospray at 3500 Volts and

positive scan mode, nebulizer gas at 

approximately 10 L/min flow

DETECTOR(MS/MS): Dwell time=100-200 msec

 Collision gas: Nitrogen or Argon

MULTIPLE REACTION MODE: 

Quantification mass transitions:

Mitomycin C = m/z 335 → 242, Porfiromycin = 349 → 256

TOTAL RUN TIME: Approximately 24 minute cycle time 

CALIBRATION: Standards from Mitomycin C solutions with 

internal standard

QUALITY CONTROL: At least one level of spiked sample prepared 

from a separately weighed stock solution

RANGE: 20 to 5000 ng/100 cm2

ESTIMATED LOD: LOD is dependent upon column and 

instrumentation. Instrumental LOD is equivalent 

to approximately 2 ng/100 cm2

; by lowest 

standard level LOD is equivalent to 20 ng/100 

cm2

PRECISION (SS�rr): See Table 1

APPLICABILITY: Mitomycin C is an antineoplastic drug often used in the operating room setting. This method measures the 

quantity of the target analyte from surface contamination. The method was tested on three “hard” surfaces likely to be found in 

operating rooms. (See Table 1.) 

INTERFERENCES: None found or identified. 

OTHER METHODS: This method is from the one described by B’Hymer [2,3]. There are numerous literature methods for this 

compound, but standardized methods from governmental or other agencies are lacking.
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 2 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

REAGENTS:

1. Mitomycin C (CAS no. 50-07-7) reference 

standard 0.10 mg/mL stock solution in 

acetonitrile. Store in a freezer at typical 

temperatures (-10 to -20 °C)

2. Porfiromycin (CAS no. 801-52-5) reference 

standard 0.05 mg/mL stock solution in 

acetonitrile. Store in a freezer at typical 

temperatures (-10 to -20 °C)

3. Porfiromycin spiking solution made 0.25 

μg/mL in acetonitrile. Store in a freezer at 

typical temperatures (-10 to -20 °C)

4. Acetonitrile, HPLC grade or better*

5. Acetic acid, glacial, HPLC grade or better*

6. Ammonium hydroxide, ACS grade or 

better*

7. Citric acid, monohydrate, ACS grade or 

better

8. Isopropanol, HPLC grade or better*

9. Water, HPLC grade or better

10. Autosampler needle rinse solution, 50/50% 

(v/v) acetonitrile/water

11. Extraction solvent 20/45/35% (v/v/v) 

acetonitrile /isopropanol/water. Mix the 

components and then add enough citric 

acid monohydrate to make 0.01 M. Adjust 

the apparent pH of the mixture to 7.0 by the 

addition of ammonium hydroxide. 

12. Mobile phase A (10/90/0.1%, v/v/v) 

acetonitrile/water/acetic acid, filtered

13. Mobile phase B (75/25/0.1%, v/v/v)

acetonitrile/water/acetic acid, filtered

14. Nitrogen, UHP or better

*See SPECIAL PRECAUTIONS.

EQUIPMENT:

1. High-performance liquid chromatograph 

(HPLC) equipped with a tandem mass 

spectrometric detector with data 

collection system

2. HPLC column (C18, 150 x 3 mm, 3.5 μm 

particle size)

3. HPLC guard column (C18, 4.0 x 2.0 mm)

4. Autosampler capable of maintaining 8 °C 

5. Pre-column filter, 2 μm 

6. Pipettor with disposable tips in the 1000 

μL volume delivery range

7. Chemical orbital shaker

8. pH meter

9. Filter paper disc, 55 mm, Grade 42

10. Polyester swab, 13 mm head width

11. Filter discs, polyvinylidene difluoride 

(PVDF), 0.22 μm 

12. Polypropylene jar, 125 mL wide mouth

13. Polypropylene tubes, 15 mL 

14. Metal spatula

15. Volumetric flasks, glass: 10, 50, 100 and 

250 mL

16. Autosampler vials, amber glass, with caps 

and septa

17. Disposable glass pipets

18. Sintered glass filtering apparatus and 1-L 

side- armed Erlenmeyer flask

19. Filters, glass microfiber, size to fit filtering 

apparatus, 0.7 μm 

20. Analytical balance, to ± 0.00001 g

21. Template: 10 cm x 10 cm hole. Template 

made of relatively rigid disposable 

cardstock (highly recommended) or sheet 

of PTFE.

22. Wire cutters or other device capable of 

snipping the heads off the swabs. 

23. Gloves, chemotherapy-safe, powderless

24. Waste containers, chemotherapy 

approved

 

SPECIAL PRECAUTIONS: Acetic acid, acetonitrile and isopropanol are flammable; handle with care and 

use in a chemical fume hood. Handle all chemicals using the required safety precautions. Reagents with 

manufacturer expiration dates should be observed. Mitomycin C and porfiromycin are carcinogens [4] and 

should be handled and disposed of properly [5,6]. 
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 3 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

SAMPLING:

1. Identify the surface area to be sampled. Wearing a clean pair of powderless, chemotherapy-safe

gloves, place a 10 x 10 cm (100 cm2

) template on the surface and secure with tape. If the template

cannot be placed on the targeted surface, measure 100 cm2 and outline with tape. For irregular shaped

objects, measure the length and width in order to calculate and record the surface area.

NOTE: Mitomycin C does show some light sensitivity (see Stability under Evaluation of Method

section), so reasonable steps to minimize light exposure should be taken.

2. Apply 250 μL of extraction solvent to the area inside the template by use of an automatic pipet,

moving it over the surface in order to spread the solvent as much as is possible. The surface can be

wiped immediately. Reapply an aliquot of extraction solvent within the template for each wiping

instance.

NOTE: If the surface to be sampled is not amenable to the application of the extraction solvent (such as

a wall or doorknob/handle or other surfaces where the solvent would run off), the solvent may 

be applied to the swabs or filter paper and used to wipe. Apply 250 μL of the extraction solvent 

to each of the three swabs or wipes in this case.

3. If using the polyester swab procedure:

a. Apply even pressure with the swab and wipe using an overlapping S-pattern in the horizontal

direction within the template. Invert the swab exposing the unused side, then wipe the same

surface using an overlapping S-pattern in the vertical direction. Ensure that you have wiped the

entire area inside the template. Cut, snip, or break the swab at the indentation above the

swabbing material and drop into the 125-mL wide-mouth jar.

NOTE: The wide-mouth jar should be pre-labelled.

b. Repeat the process two more times and combine all three swab wipes into the wide-mouth jar.

NOTE: Gloves and the template are not to be changed during these repeated samplings of the

same area.

c. Record the time, date, location and condition of the area that was sampled along with the name of

the analyst performing the sampling. Photographs are useful when evaluating surface results.

4. If using the alternative filter paper procedure:

a. Grasp the opposite edges of the filter paper disc producing a flat area with which to wipe the

surface. Apply even pressure with the paper disc and wipe using an overlapping S-pattern in the

horizontal direction within the template. Then wipe the same surface with the same side of the 

paper disc using an overlapping S-pattern in the vertical direction. Ensure that you have wiped 

the entire area inside the template. Use caution not to touch the area being sampled with your 

gloved hand while collecting the sample. Place the filter paper in the 125-mL wide-mouth jar. 

NOTE: The wide-mouth jar should be pre-labelled.

b. Repeat the process two more times and combine all three filter wipes into the wide-mouth jar.

NOTE: Gloves and the template are not to be changed during these repeated samplings of the

same area.

c. Record the time, date, location and condition of the area that was sampled along with the name of

the analyst performing the sampling. Photographs are useful when evaluating surface results.

5. Remove gloves and discard into trace/residual chemotherapy waste containers, often known as the

yellow containers in hospital settings (or follow applicable guidelines [5,6]). Wear a new pair of clean

gloves for each new sample.

6. Use a new disposable template for each new sample. The used template can be discarded discard into

trace/residual chemotherapy waste containers, often known as the yellow containers in hospital

settings (or follow applicable guidelines [5,6]). The use of disposable templates is recommended as it is

very difficult to properly clean reusable templates, especially in a field setting

7. Wash hands thoroughly after sampling is completed.
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 4 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

8. To collect field blanks, remove unexposed wipes (3) or swabs (3) from their packaging and place into

pre-labelled 125-mL wide-mouth jars. Add 750 μL of extraction solvent to the field blanks to mimic the

volume used in the sampling procedure. If desired, the jars may be secured with a sealing film or tape

before shipment.

9. Ship the wide-mouth jars in a packing container that is designed for dry-ice storage and transport.

Ship express for delivery to the laboratory.

10. Upon receipt in the laboratory, inventory, and store the samples in a freezer at typical temperatures

(-10 to -20 °C or colder) until the time of analysis.

SAMPLE PREPARATION:

11. Remove the sample jars from storage and add extraction solvent to make 9.0 mL of volume [0.75 mL

was used to wipe the surface, so 8.25 mL would be added assuming minimum evaporation from the

jar].

12. Add 1.0 mL of the porfiromycin internal standard spiking solution to the sample jar.

NOTE: At this point, the swab heads or filter paper discs should be covered in solvent.

13. Tightly cap the jar and place on the orbital shaker for swirling. Swirl for 30 minutes at 100 rpm.

14. Remove and filter an aliquot of the sample through the 0.22 μm PVDF filter disc into a 15-mL

polypropylene tube.

15. Transfer approximately 1 mL of each filtered sample solution into an amber HPLC autosampler vial.

Protect from light sources.

CALIBRATION AND QUALITY CONTROL:

16. Prepare a mitomycin C dilute standard solution by diluting 5.0 mL of the stock solution to 100 mL with

acetonitrile (approximately 5 μg/mL mitomycin C concentration).

17. Mitomycin C standard solutions. The 5 μg/mL concentration solution is diluted in extraction solvent to

make 2, 5, 10, 50, 100, 200, 400 and 500 ng/mL mitomycin C solutions.

18. Transfer 1.0 mL of each mitomycin C solution from step 17 into separate HPLC autosampler vials.

19. Add 0.100 mL of the internal standard spiking solution to each vial. These standards are equivalent to

the extract solutions which would result for wiping surfaces prepared at levels of 20 to 5000 ng/100

cm2 mitomycin C.

20. Prepare at least one blank sample of the extraction solvent without analyte spikes or internal standard

and at least one extraction solvent blank with 0.100 mL internal standard added.

21. Prepare at least one quality control (QC) standard of mitomycin C using separately prepared stock

solutions. A 50 ng/mL equivalent spike level is suggested, and more than one level can be used if

desired. Run enough QC samples to be greater than 5% of the sample set. QC values should normally

be within ±20% of the spiked values. If not, the batch is considered out of control, the batch data

discarded, and corrective actions taken before more specimens are analyzed.

MEASUREMENT:

22. Set the high-performance liquid chromatograph according to the manufacturer’s recommendations

and to the conditions listed on the first page of this method. Following each injection, a needle rinse

with 50/50% (v/v) acetonitrile/water is required to eliminate sample carry-over by the autosampler.

23. Set the mass spectrometric detector to multiple reaction mode (MRM) according to the manufacturer’s

recommendations and the conditions listed on the first page of this method. Example conditions are

summarized below and should be optimized for a specific instrument for mitomycin C and

porfiromycin:
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 5 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

MS/MS conditions using positive electrospray ionization and nitrogen as the collision gas

Analyte Parent ion

(m/z) 

MS1 

Resolution

Daughter

ion (m/z) 

MS2 

Resolution

Dwell

(ms)

Fragmentor

(V)

Collision

Energy 

(V)

Cell

Voltage 

(V)

Mitomycin C 335 unit 242 unit 200 97 8 4 

Porfiromycin 349 unit 256 unit 200 97 6 4 

MS/MS conditions using positive electrospray ionization and argon as the collision gas (mass span at 

1.0)

Analyte Parent ion (m/z) Daughter ion (m/z) Dwell (ms) Cone (V) Collison Energy (V)

Mitomycin C 335 242 100 17 15 

Porfiromycin 349 256 100 17 16 

NOTE: Differences in detector parameters and their optimal values reflect use of different models of MS 

detectors. Models used can be found in Tables 1 and 2 and the method backup data report [2].

24. Inject 5 μL of each sample extract, field blank, standard, QC standard, and blank. A sample

chromatogram is shown in Figure 1.

25. Measure the peak areas of the mitomycin C analyte and that for the internal standard in the

chromatograms. Divide the peak area of the analytes by the peak area from the internal standard (IS).

26. Prepare calibration curves of the peak area mitomycin C/Area IS versus the concentration of the

standards for the analyte. The standard curve should have a coefficient of determination (r2

) of equal

to or greater than 0.98 to be acceptable for use. Furthermore, when each standard is plugged back

into the calibration equation, the value should be within ±20% of the expected.

CALCULATIONS:

27. Determine the concentration of the mitomycin C in the extracts from the surface area (10.0 mL

solution for the 100 cm2 wiped surface area) from the curves obtained in step 26. The results can be

expressed as ng/100 cm2 mitomycin C.

EVALUATION OF METHOD:

This method was evaluated and described in detail by B’Hymer [2,3]. A general summary of this previously 

published information is given below.

Accuracy and Precision. Several recovery studies using two columns over several days demonstrated the 

accuracy and precision of this test method. These matrix matched samples were made to mimic matrices 

that would be found in the field. Blank surfaces were wetted and the surfaces sampled as in Steps 3 or 4 of 

the Sampling Section. The solutions containing the three wipes (filter paper or swab heads) were then 

spiked with varying amounts of the analyte and carried through the analytical process. A primary recovery 

experiment using the optimized conditions with filter paper matrix matched solution fortified with 

mitomycin C was performed to demonstrate the general accuracy and precision of the HPLC conditions 

using the Agilent mass spectrometer. These data are presented in Table 1. Average recoveries ranged from 

93 to 105% for the six fortified levels of mitomycin C investigated. A similar recovery study using the swab 

matrix matched solution fortified with mitomycin C was performed using the Waters Xevo mass 

spectrometer. These data are presented in Table 2. Average recoveries ranged from 90 to 107% for the four 
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 6 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

fortified levels of mitomycin C investigated. These accuracy and precision numbers fall well within 

parameters established by guidelines suggested by the US Food and Drug Administration [7].

The second recovery study was performed from surfaces with mitomycin C and these data are presented in 

Table 3. Both filter paper and Texwipe® polyester swabs were evaluated. Some general conclusions can be 

drawn from these data. Recoveries from the vinyl surface were generally lower owing to the porous nature 

of the surface. The two types of wipes worked similarly on stainless steel and showed less than quantitative 

recoveries at the two lowest concentrations tested. The polyester swab gave higher recoveries and greater 

precision than that of the filter paper when used on the Formica surface at the lower concentrations. 

Similar to the stainless steel, recoveries at the two lowest concentrations still average less than 75%.

The two high concentration points (10 and 25 μg/100 cm2

) studied for both recovery studies were outside 

the standard calibration curve range. For these levels, a 1 to 10 dilution of the filtered extract solution was 

performed to demonstrate that the method was still accurate at higher levels of mitomycin C surface 

contamination. 

Linearity. All calibration curves used during the development of this method were linear, had correlation 

of determinations (r2

) of 0.99 or greater, and had y-intercepts near zero. The concentration range was 

equivalent to 20 to 5000 ng/100 cm2 mitomycin C surface contamination. Calibration curves were run at 

the beginning and end of all sample batch runs; calibration curve slope drift was found to be minimal 

during this method’s development.

Specificity. The optimized chromatographic conditions developed for this method, along with the 

tandem mass spectrometric detector, proved to be specific and have no major interferences. The mass 

transition ion selected for mitomycin C had the greatest response and the predominant daughter ion was 

chosen.

Robustness. Two Zorbax Rx-C18 HPLC columns of different manufacturing lots were used during the 

recovery studies. Accuracy and precision did not appear to be affected; therefore, the method appears to 

be reproducible with any functioning Zorbax Rx column. Two HPLC-MS/MS systems were investigated for 

use with this method and found to give reasonable recovery results. 

Stability. Mitomycin C is a pharmaceutical product, and as such, its stability in solution has been well 

established in the chemical literature. It can be easily hydrolyzed under acidic or basic conditions, but it is 

generally stable in a pH range between 7 and 8. The acetonitrile-isopropanol-water ammonium citrate

system developed for the current method was found to be a good matrix for sample stability, and it was 

experimentally determined to be superior to the propylene glycol-water mixtures reported in patented 

pharmaceutical formulations [2,8]. Mitomycin C solutions stored in the acetonitrile-isopropanol-water 

extraction solvent at room temperature in darkness and those held at 8 oC in the autosampler were found 

to degrade only 3 to 4% after 8 days (n=3); the optimized extraction solvent also gave excellent recoveries 

during the surface wipe testing. Stock solutions of mitomycin C stored in 100% acetonitrile were found to 

degrade only 2 to 3% after 3 to 6 months of storage at -15 oC. Mitomycin C did show some light sensitivity. 

Sample solutions (n=3) made in the extraction solvent degraded 12% after 8 days exposed to window 

sunlight and the laboratory fluorescent lights. From these experiments, it was deemed advisable to make 

an effort to minimize light exposure of mitomycin C solutions. Amber autosampler vials were used for all 

recovery studies while developing this method. While no sample holding time studies were performed, 

two pieces of information would seem to be relevant in this matter. The first is that solid mitomycin C is 

known to be stable for at least one year when stored frozen [9-11]. The second is that solutions of 

mitomycin C in the extraction solvent have been shown to be stable for greater than 8 days at 8 oC [2]. 

Collected samples are dissolved in the extraction solvent on the surface of the filter paper or swab and 

then frozen. One can safely state that the samples would be expected to be stable for at least 8 days and 

since they are frozen and more like their solid state, it would be expected that they would be stable for 

much longer timeframes.
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 7 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

Range. This method should be considered accurate for the estimation of mitomycin C surface 

contamination within the standard curve range. Field samples at higher levels can be diluted to a 

concentration within that range for analysis.

REFERENCES:

[1] The Merck Index [2013]. The Merck Index, 15th Edition. The Royal Society of Chemistry: Cambridge,

UK.

[2] B’Hymer C and Connor TH [2014]. Backup data report, Draft NIOSH Method 9212.

[3] B’Hymer C, Connor T, Stinson D, Pretty J [2015]. Validation of an HPLC-MS/MS and wipe procedure

for mitomycin C contamination. J Chromatogr Sci 53:619-624.

[4] IARC [1975]. IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Man:

Some naturally occurring substances. Vol. 10. International Agency for Research on Cancer, Lyon,

France.

[5] USP [2016]. USP General Chapter <800> Hazardous drugs – handling in healthcare settings. The

United States Pharmacopeial Convention, Rockville, MD,

https://www.usp.org/compounding/general-chapter-hazardous-drugs-handling-healthcare

[6] EPA [1976]. Resource Conservation and Recovery Act (RCRA), Part 266, Subpart P. The United States

Environmental Protection Agency, Washington, DC. www.ecfr.gov/cgibin/retrieveECFR?gp=&SID=c94567294dff611654af7a3944a91d69&mc=true&r=PART&n=pt40.29.26

6#sp40.29.266.p

[7] FDA [2018]. Guidance for industry: bioanalytical method validation. U.S. Food and Drug

Administration, Rockville, MD. www.fda.gov/media/70858/download.

[8] Velpandian T, Saluja V, Ravi AK, Kumari SS, Mathur R, Biswas NR, Ghose S [2005]. Evaluation of the

stability of extemporaneously prepared ophthalmic formulation of mitomycin C. J Ocul Pharmacol

Ther 21:217-222.

[9] Enzo Life Sciences [2018]. www.enzolifesciences.com/BML-GR311/mitomycin-c.

[10] BPS Bioscience [2018]. www.bpsbioscience.com/mitomycin-c.

[11] Cayman Chemical [2018]. www.caymanchem.com/pdfs/11435.pdf.
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 8 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

organizations or their programs or products. Furthermore, NIOSH is not responsible for the content of

these websites. All web addresses referenced in this document were accessible as of the publication date.

FIGURE 1

Chromatogram of a sample taken from a spiked matrix solution. The mitomycin C and porfiromycin

solution concentrations are approximately 20 ng/mL.

METHOD WRITTEN (REVISED) BY:

Clayton B'Hymer, PhD, NIOSH/HELD/CBMB and Thomas H. Connor, PhD, NIOSH/DSI/REB. Final editing 

performed by Dale A. Shoemaker, PhD, NIOSH/HELD/CBMB.

Disclaimer: Mention of any company or product does not constitute endorsement by the National 

Institute for Occupational Safety and Health, Centers for Disease Control and Prevention. In addition, 

citations to websites external to NIOSH do not constitute NIOSH endorsement of the sponsoring 
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 9 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

Table 1

Recovery of Mitomycin C Spikes Added to Filter Paper Matrix-Matched Extracts

(Analysis was performed using an Agilent Model 6430 Mass Spectrometer)

Surface Equivalent Level

(μg/100 cm2

) 

Mean

(n=10) 

% Recovery % RSD 

(n=10) 

Stainless Steel 0.100 0.104 104 7.5

0.200 0.202 100 3.3

2.00 2.01 100 1.0

4.00 4.08 102 2.7

10.0 9.81 98 2.5

25.0 23.2 93 5.1

Vinyl 0.100 0.105 105 3.0

0.200 0.201 101 2.3

2.00 1.99 100 3.1

4.00 4.06 102 2.6

10.0 9.82 98 2.7

25.0 23.8 95 5.9

Formica® 0.100 0.103 103 3.3

0.200 0.200 100 2.7

2.00 2.02 101 3.0

4.00 4.05 101 1.8

10.0 9.66 97 2.0

25.0 23.6 94 5.4

Notes: This recovery study was performed using two different Zorbax columns. RSD is relative standard 

deviation.
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 10 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

Table 2

Recovery of Mitomycin C Spikes Added to Texwipe® Swab Matrix-Matched Extracts

(Analysis was performed using a Waters Xevo TQ Mass Spectrometer)

Surface Equivalent Level

(μg/100 cm2

) 

Mean

(n=5) 

% Recovery % RSD 

(n=5) 

Stainless Steel 0.100 0.090 90 1.3

0.200 0.187 94 1.4

2.00 2.02 101 4.1

4.00 4.20 105 2.1

Vinyl 0.100 0.105 105 2.4

0.200 0.200 100 1.3

2.00 1.99 99 1.0

4.00 3.96 99 1.1

Formica® 0.100 0.107 107 1.7

0.200 0.214 107 0.8

2.00 2.06 103 1.2

4.00 4.21 105 0.8
Mitomycin C on Surfaces: METHOD 9212, Issue 1, dated 30 May 2020 - Page 11 of 11 

NIOSH Manual of Analytical Methods (NMAM), Fifth Edition

Table 3

Recovery from Spiked Material Surfaces

Surface Filter Paper Wipe Polyester Swab

Level

(μg/100 cm2

) 

Mean

(n=5) 

% Recovery % RSD Level

(μg/100 cm2

) 

Mean

(n=5) 

% Recovery % RSD 

Stainless Steel 0.100 0.068 68 6.3 0.100 0.071 71 6.4

0.200 0.124 62 10.8 0.200 0.122 61 5.6

2.00 1.75 88 3.6 2.00 1.77 89 3.6

4.00 3.67 92 3.8 4.00 3.76 94 2.5

10.0 9.80 98 8.3 10.0 9.76 98 1.6

25.0 23.9 96 3.1 25.0 24.8 99 2.6

Vinyl 0.100 0.058 58 7.9 0.100 0.062 62 6.2

0.200 0.115 58 12.6 0.200 0.109 54 3.7

2.00 1.12 56 7.9 2.00 1.32 66 4.3

4.00 2.02 51 8.0 4.00 2.39 60 5.9

10.0 6.25 63 3.4 10.0 5.32 53 8.2

25.0 14.8 59 7.6 25.0 14.0 56 5.1

Formica® 0.100 0.030 30 47 0.100 0.073 73 7.9

0.200 0.065 33 6.0 0.200 0.126 63 5.3

2.00 1.53 77 8.5 2.00 1.74 87 1.2

4.00 3.27 82 1.2 4.00 3.65 91 1.7

10.0 9.05 91 2.0 10.0 9.26 93 2.9

25.0 24.0 96 3.7 25.0 24.3 97 0.7

Note: These data were collected using the Agilent HPLC-MS/MS system.