This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Marketing Notification No. 13A2X00197218106**Revised: March 2017 (6th edition)*Revised: January 2017 (5th edition)

This package insert must be read carefully prior to use.
Theophylline assay kit(Classification No.: 30417000)

patients who have not previously shown even mild
symptoms of toxicity, monitoring of the blood level oftheophylline is important during treatment. The rate ofclearance of theophylline from the blood may alsovary due to other factors. For example, clearance fromthe blood is delayed in obese patients, patients withliver disorders, and patients on a highcarbohydrate/low-protein diet. It is known thatclearance of theophylline from the blood is very slow
in premature infants,1） but is rapid in smokers.2）
Therefore, monitoring the blood level of theophylline
along with consideration of other clinical data is usefulfor adjusting the dosage of this drug so that theoptimum clinical response can be obtained without
symptoms of toxicity.

Nanopia TDM Theophylline
General Precautions
1. This product is for in vitro diagnostic use, and mustnot be used for any other purposes.
2. Clinicians should make a comprehensive clinicaldecision based on assay results in conjunction withclinical symptoms and other examination results.
3. This product should be used only as directed in thispackage insert. Reliability of results cannot be
guaranteed if there are any deviations from theinstructions in this package insert.
4. If the reagent accidentally comes in contact witheyes and/or mouth, rinse immediately with ample
water as first aid, and consult the doctor if required.
5. Carefully read the operating instructions for eachtype of automated analyzers prior to using thisproduct. Parameters for each type of analyzers areavailable, and can be requested from SEKISUIMEDICAL CO., LTD. if required.
6. Perform a quality control test prior to assay toensure accuracy.

Assay Principle
1. Assay Principle
When a certain amount of anti-theophylline
antibody is added and reacted with a sample,
consumption of the antibody depends on its content
in the sample. When theophylline-coated latex isadded, residual anti-theophylline antibody reactswith the latex and forms aggregates.
Since the extent of aggregation depends on thetheophylline concentration in the sample, thetheophylline concentration can be determined bymeasuring aggregation as the change ofabsorbance.

Description (Kit Components)
Component: Ingredients
THE Antibody Solution 1:
Anti-theophylline mousemonoclonal antibody
THE Latex Reagent 2:
Theophylline-coated latex

Sample (theophylline) + Anti-theophylline
antibody ──────▶ Antigen-antibody reaction
Unreacted
anti-theophylline
antibody+Theophylline-coated latex───▶ Aggregation by antigen-antibody reaction

Intended Use
Measurement of theophylline in serum or plasma

2. Features
1) Because a highly specific monoclonal
antibody is used, this product shows excellent
sensitivity and accuracy.
2) Liquid reagents, ready-to-use.
3) Applicable to various automated analyzers.

Theophylline (1,3-dimethylxanthine) is a methyl
xanthine derivative that is widely used for thetreatment of bronchial asthma, asthmatic bronchitis,and neonatal apnea. The main action of theophylline isrelaxation of bronchial smooth muscle. It also
activates the central nervous system and therespiratory center in the medulla oblongata, reduces
peripheral vascular resistance, stimulates the heart,and causes diuresis.
The clinical effects of theophylline are closely
correlated with its blood level. Symptoms oftheophylline toxicity usually occur at a concentration> 20 µL/mL in adults.2 However, mild symptoms oftoxicity may develop at a concentration > 15 μg/mL,depending on the person. Mild symptoms of toxicity
include anorexia, nausea, vomiting, headache, andnervousness. Severe symptoms of toxicity include anincreased heart rate, arrhythmia, cerebral seizure,respiratory arrest, and cardiac arrest. These symptoms
occur at concentrations > 40 µg/mL, but also at a lower
concentration in some patients.
Clearance of theophylline shows individual variation.
Because severe symptoms of toxicity may occur inProcedural Precautions ＊1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma, EDTA
plasma and citrated plasma) may be used.
2) Storage of samples
If serum or plasma samples cannot be measured
on the day of separation, store them as follows.
Avoid repetition of freezing and thawing.
2–8C: for tests within 7 days≤ -20C: for tests within 3 months
Bring samples to room temperature (15–30C)
before use.
3) Caution must be exercised, because aseparating agent, etc. in the blood collection
tube may affect assay values.4)14) Sampling should be performed after
removing insoluble matter from the sample.
Highly turbid specimens should becentrifuged before assay.
2. Interfering substances
1) Assay results are not affected by free bilirubin(up to 20 mg/dL), conjugated bilirubin (up to 20mg/dL), hemoglobin (up to 500 mg/dL),ascorbic acid (up to 50 mg/dL), formazinturbidity (up to 2500 FTU), or rheumatoid
factors (up to 450 U/L).
2) Because mouse antibody is used in the assay,
artifactual elevation of results may occur if thesample contains human anti-mouse antibody. In
this case, perform re-measurement by anothermethod.
3) Cross-reactivity
The following table summarizes drugs related
to theophylline and cross-reactivity between
theophylline and other drugs.

Substance
1,3,7-Trimethyluricacid
1,3-Dimethyluric acid
1,7-Dimethyluric acid
1,7-Dimethylxanthine
1-Methyluric acid
1-Methylxanthine
3,7-Dimethyluric acid
3-Methyluric acid
3-Methylxanthine7-(β-hydroxyethyl)theophylline
7-(2-Hydroxypropyl)
theophylline
7-Methylxanthine
8-Chlorotheophylline
Allopurinol
Ampicillin
Caffeine
Clindamycin
Diprophylline
Heparin
Hypoxanthine
Phenobarbital
Prednisone
Pseudoephedrine
Terbutaline
Theobromine
Urea
Uric acid
Xanthine
Xanthosine

Concentration
tested(µg/mL)
1000

Crossreactivity (%)

200
1000
1000
1000
1000
1000
1000
1430
1430

2.290.09
1.090.06
0.49
00.090.65
0.79

1818

0.58

1000
360
1000
2000
645
2000
2000
2000
1000
2000
2000
2000
2000
800
2000
1000
1000
1000

0.05
1.34
0.01
0.04
2.60
0.05
0.46
000.03
00.01
0
1.85
0
0.03
0.03
0.03

(manufactured by SEKISUI MEDICAL CO.,LTD.), and perform re-measurement.
Dosage/Administration (Assay Procedure) ＊1. Preparation of reagents
Reagent (1): THE Antibody Solution 1 is readyto use.
Reagent (2): THE Latex Reagent 2 is ready touse.
Before using this product, gently invert the THE
Latex Reagent 2 bottle to mix it thoroughly, andcheck that there are no bubbles.
2.
Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.
37C37CReagent
Sample Reagent
Measurement+(1)
(2)
2 μL
270
70 (Absorbance I※)
180 μL
30 μL
sec
sec
37CMeasurement
247 (Absorbance II※)sec
Calculation ofconcentration

0.02※

Absorbance I and II: Absorbance at 700 nm

Calibration material: TDM Calibrator for Nanopia(manufacture's assigned value)＊＊

Assessment of Assay Results ＊1. Reference standard range
The effective concentration of theophylline is 5–15μg/mL5) in adults and 5–10 μg/mL6) in infants.
However, the effective concentration range may
overlap with the toxic concentration range.
Therefore, interpretation of assay results should
also be based on the patient's clinical findings andother examination results.
It is recommended that the blood concentration oftheophylline should be measured in a sufficientnumber of samples for statistical analysis and thatits clinical effective concentration should bedetermined by each medical institution.
2. There may be reactions or interfering reactions
with non-target substances. If assay results appear
to be unreliable, repeat the measurement (ifnecessary, after dilution) or try another analyticalmethods.
Performance
1. Sensitivity
1) The change of absorbance of the standard
solution (0 μg/mL) per minute is 0.145–0.300.
2) The ratio between the change of absorbance perminute with the standard solution (0 µg/mL)
and that with another standard solution (2.5µg/mL) is 45–85 %.
2. Accuracy: 80–120 % of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 10 %(Test methods used for 1.–3. are in-house methods.)
4. Measurement Range7): (On Hitachi 7170S
automated analyzer)

3. Others
1) Always use TDM Calibrator for Nanopia
for calibration.
2) Precautions for assay range
If the concentration of a target substance in thesample exceeds the measurement range, dilutethe sample with a separately sold diluent20.2–40 μg/mL
5. Correlation7)
1) Serum N=60
r=0.993y=1.05x+0.51
Control method: Approved in vitro diagnostic(enzyme immunoassay)
2) Plasma N=77
r=0.999
y=1.06x–0.45
Control method: Approved in vitro diagnostic(enzyme immunoassay)
6. Standard Material
Theophylline (U.S. Pharmacopoeia)(The expiration date is printed on the outerpackage.)
Packaging
NamePackage
THE Antibody
1 × 36 mL
Nanopia
Solution 1
TDM
THE Latex Reagent
Theophylline
1 × 6 mL
2Constituent reagents are available in otherconfigurations. For further details please contact
SEKISUI MEDICAL CO., LTD.

Precautions for Use or Handling ＊1. Precautions for Handling (to Ensure Safety)
1) All samples used in the test should be handled asa material possibly infected with HIV, HBV,HCV, or other viruses. To prevent infection, use
disposable gloves and avoid mouth pipetting
during the test.
2) Sodium azide is added as an antiseptic agent in
the THE Antibody Solution 1 and THE Latex
Reagent 2. Therefore, if the reagent comes in
accidentally contact with eyes, mouth or skin,rinse immediately with ample water as first aid,
and consult the doctor if required.
2. Precautions for use
1) This product should be stored as directed,without freezing. Freezing can deteriorate thereagents, which can produce inaccurate results.
Therefore, avoid using the reagents which havebeen previously frozen.
2) Do not use expired reagents. Use of suchreagents cannot guarantee the reliability ofmeasurement values.
3) Do not replenish the reagents.
4) Do not mix materials from different kit lotnumbers.
5) Do not perform the assay under direct sunlight
3. Precautions for Disposal
1) Before disposal, used samples and theircontainers must be immersed in sodium
hypochlorite solution at a concentration ofgreater than 0.1% for longer than 1 hour or
autoclaved at 121ºC for 20 minutes.
2) To prevent infections from spilled samples orsolutions containing samples, wipe the spilledarea thoroughly with disinfectants such as
sodium hypochlorite solution at a concentrationof greater than 0.1%.
3) The reagents and treated samples should bediscarded as medical waste or industrial waste
according to the waste disposal regulations.
4) The reagents should be disposed of inaccordance with the Water Pollution Control act
or related regulations.
5) Sodium azide has been added as an antiseptic
agent in the THE Antibody Solution 1 and THE
Latex Reagent 2. It can react with lead or copper
pipes to produce the highly explosive metal azide.
Therefore, the reagent should be flushed withlarge amounts of water during disposal.
4. Other precautions
Do not use the containers for other purposes.

References ＊1) Ogilvie R. I. ： Clinical pharmacokinetics oftheophylline. Clinical Pharmacokinetics, 3 ：267−293, 1978.
2) Hendeles L., Weinberger M. M. ： Theophylline
therapeutic use and serum concentration
monitoring, in Taylor, W. J., Finn, A. L（. eds）：
Individualizing Drug Therapy ：PracticalApplications of Drug Monitoring, Gross
Townsend Frank, Inc., New York, vol. 1, 31−66,1981.
3) Aranda J. V., et al. ： Pharmacokinetic aspects oftheophylline in premature newborns, N. Engl. J.
Med., 295： 413−416, 1976.1)
4) Sawada T. et al.: J Med Pharm Sci, 51(1), 131−141,2004.
5) Japanese Society of Allergology: Asthma
prevention and management guidelines 2015, 116–136, Kyowa Planning, 2015.
6) Ministry of Health, Labour and Welfare,
Pharmaceutical and Food Safety Bureau, Safety
Division: Guidelines for the appropriate use ofdrugs for infantile bronchial asthma, 13, 2006.
7) In house data, SEKISUI MEDICAL CO., LTD.
Contact
SEKISUI MEDICAL CO., LTD.
international@sekisui.com
Manufacturer ＊SEKISUI MEDICAL CO., LTD.
1-3, Nihonbashi 2-chome, Chuo-ku, Tokyo, Japan

Storage and Shelf Life ＊1. Storage temperature: 2–8C
2. Shelf life: 16 months from the date of manufacture3