This package insert follows the Pharmaceuticals, Medical devices and Other Therapeutic Products Act of Japan.
In Vitro Diagnostics
Certification No. 227ADAMX00073000**Revised: December 2019 (4th edition)*Prepared: April 2017 (3rd edition)

This package insert must be read carefully prior to use.
Zonisamide assay kit(Classification No.: 42928000)

this drug states that measurement of the bloodconcentration is recommended for adjusting thedosage more appropriately.1)

Nanopia TDM Zonisamide

Assay Principle
1. Assay Principle
When a certain amount of anti-zonisamide
antibody is added and reacted with a sample,
consumption of the antibody depends on its content
in the sample. When zonisamide-coated latex isadded, residual anti-zonisamide antibody reacts
with the beads and forms aggregates. Since theextent of aggregation depends on the zonisamide
concentration in the sample, the zonisamideconcentration can be determined by measuring
aggregation as the absorbance.

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. Please read carefully the "PRECAUTIONS" section,
particularly under "Effects on Laboratory Tests" ofthe package insert of each drug, for information on
the influence to the assay result that medications
administered to the patient have. Please also read
carefully the "2) Cross-reactivity," under "2.
Interfering Substances," in the "Procedural
Precautions" section, as well as "2. Precautions forAssessment" in the "Assessment of Assay Results"
section, of this package insert.
4. This product should be used only as directed in thispackage insert. Reliability of results cannot beguaranteed if there are any deviations from theinstructions in this package insert.
5. 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.
6. 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.
7. Perform a quality control test prior to assay toensure accuracy.
8. ZNS Antibody Solution 1 contains human-derived
components that have been confirmed to be
negative for HBs antigens, HIV antibodies, andHCV antibodies. However, handle this reagent withgreat care in the same manner as the samples,because it is potentially infectious.

Sample (zonisamide) + Anti-zonisamide antibody───▶ Antigen-antibody reaction
Unreacted
anti-zonisamide
antibody+Zonisamide-coated latex beads───▶ Aggregation by antigen-antibody reaction2. Features
1) Liquid reagents, ready-to-use.
2) Applicable to various automated analyzers.
Procedural Precautions **1. Properties of Samples and Sampling Methods
1) Samples
Serum and plasma (heparin plasma andEDTA plasma) may be used.
2) Storage of samples(1) After separation of serum (plasma),samples may be stored for up to 14 days ina refrigerator. If samples cannot bemeasured within 14 days of serum (plasma)separation, store them at −20C or lowerand measure them within 1 month. Storedsamples should be brought to room
temperature (15–30C) before use.
(2) Samples may be frozen and thawed up totwice.
2. Interfering substances
1) Assay results are not affected by free bilirubin(up to 18.9 mg/dL), conjugated bilirubin (up to20.8 mg/dL), hemoglobin (up to 500 mg/dL),
formazine turbidity (up to 1410 FTU), or
rheumatoid factors (up to 500 IU/mL).
2) Cross-reactivity
The following table summarizes the crossreactivity with the zonisamide metabolites andother drugs.
(1) Zonisamide metabolites
If each metabolite is at the concentration
mentioned in the table below when
zonisamide-free samples and samples
containing zonisamide at low and highconcentrations are measured, crossreactivity is as follows.

Description (Kit Components)
Component: IngredientsZNS Antibody Solution 1:
Anti-zonisamide rabbit polyclonal
antibody
ZNS Latex Reagent 2:
Zonisamide-coated latex
Intended Use
Measurement of zonisamide in serum or plasma
Zonisamide is an antiepileptic drug that is used for thetreatment of partial, generalized, and mixed seizures inpatients with partial or generalized epilepsy. Adverse
reactions to this drug include sleepiness, anorexia, andataxia. Serious adverse reactions that have beenreported include toxic epidermal necrolysis (TEN),
hypersensitivity syndrome, etc. The package insert for1

2. Assay Procedure
This product is compatible with various types ofautomated analyzer. An example of the assay
procedure is indicated below.

Cross-reactivity (%)
Metabolite

Concent- ration
tested(µg/mL)

N-Acetyl
zonisamide
(NAZ)

500
100
50255

Zonisamide-freesample(0 μg/mL)

Samplecontaining alow
zonisamide
concentration(12 μg/mL)

Samplecontaining ahigh zonisamide
concentration(31 μg/mL)

0.20
ND
ND
ND
ND

0.20
ND
ND
ND
ND

0.20
ND
ND
ND
ND

37C Reagent 37C Measurement
Reagent(2)
Sample +(1) Approx. 75 μL Approx.(Absorbance
3 μL※
200 μL 5 min
1 min I )37C Measurement(AbsorbanceApprox. II※)
4 min
Calculation ofconcentration

Cross-reactivity (%)

Metabolite

Concent- ration
tested(µg/mL)

2Sulfamoyla
cetylphenol(SMAP)

500
100
50
25
5Zonisami
de-freesample(0 μg/mL)

Samplecontaining alow
zonisamide
concentration(12 μg/mL)

2.60
11.30
2.90
3.70
9.30

2.30
1.30
ND
ND
ND

Samplecontaining ahigh zonisamide
concentration(36 μg/mL)

ND
ND
4.40
5.70
4.20
ND: Not detectable※

Calibration material: Zonisamide(Manufacture's assigned value)

Clonazepam
Diazepam
Ethosuximide
Phenobarbital
Topiramate
Valproic acidAcetaminophen

Carbamazepine

Cyclosporin
Erythromycin
Ibuprofen

Concentration tested(µg/mL)
0.5
100
1000
400
250
1000
20012050200400

Drug name
Salicylic acidSulfamethoxazole
Sulfisoxazole
Caffeine
Lamotrigine
Phenytoin
PrimidoneTrimethoprim
Theophylline
HeparinLevetiracetam

Calibrator

Assessment of Assay Results
1. Reference standard range
The therapeutic range of the blood concentration ofzonisamide depends on the severity of epilepsy andvaries between individual patients. It has beenreported that the therapeutic range is 10–30 μg/mL
and the standard value may be approximately 20μg/mL. However, it has also been reported that therange is 10–40 μg/mL.2)
It is recommended that the blood concentration ofzonisamide should be measured in a sufficientnumber of samples for statistical analysis and thatits therapeutic range should be determined by each
medical institution.
2. Precautions for Assessment
There may be reactions or interfering reactions with
non-target substances. If assay results appear to beunreliable, repeat the measurement (if necessary,after dilution) or try another analytical methods.

(2) Other drugs
If each drug is at the concentration shown in thetable below when the zonisamide concentration isapproximately 12 or 36 μg/mL, cross-reactivity is≤ 2%.
Drug nameAbsorbance I and II: Absorbance at 700 nmConcentration tested(µg/mL)
500
400
1000
100300200100
202508500 U/L100※Based on the Clinical and Laboratory Standards
Institute (CLSI) EP7-A2, cross-reactivity wascalculated by the following equation.
Cross-reactivity (%) = ({[Concentration ofzonisamide in the sample after adding themetabolite or drug] – [Concentration ofzonisamide in the sample without the metabolite orconcomitant drug]} / [Concentration of themetabolite or drug in the sample]) × 100
3. Others
1) Always use Zonisamide Calibrator forcalibration.
2) Precautions for assay range
If the concentration in the sample exceeds themeasurement range, dilute the sample with theZonisamide Calibrator A and perform remeasurement.

Performance
1. Sensitivity(1) The change of absorbance per minute with thestandard solution (0.0 µg/mL) is ≥ 50 mAbs.
/min.
(2) The difference between the change ofabsorbance per minute with the standardsolution (0.0 µg/mL) and that with anotherstandard solution (80.0 µg/mL) is ≥ 35 mAbs.
/min.
2. Accuracy: 80–120 % of the expected assay value
3. Within-run Reproducibility:
Coefficient of variation ≤ 15 %(Test methods used for 1.–3. are in-housemethods.)
4. Measurement Range3) (On Hitachi 7180
automated analyzer)
3.0–80.0 µg /mL
5. Correlation3)
1) Serum
N=66
r=0.997
y=1.06x+0.22
Control method: Approved in vitro diagnostic(latex agglutination method)
2) Plasma
N=61 r=0.990
y=1.04x+0.66
Control method: Approved in vitro diagnostic(enzyme immunoassay)
6. Standard Material

Dosage/Administration (Assay Procedure)
1. Preparation of reagents
Reagent (1): ZNS Antibody Solution 1 is readyto use.
Reagent (2): ZNS Latex Reagent 2 is ready touse.
Before using this product, gently invert theZNS Latex Reagent 2 bottle to mix it thoroughly,and check that there are no bubbles.

2Zonisamide (in-house standard material)

Packaging

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 ZNS Antibody Solution 1 and ZNS 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) After completion of measurement, this productshould be tightly stoppered and stored in arefrigerator.
5) Do not mix materials from different kit lotnumbers.
6) 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 orautoclaved 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) Sodium azide has been added as an antiseptic
agent in the ZNS Antibody Solution 1 and ZNS
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.

Name
ZNS Antibody
Nanopia TDM Solution 1
Zonisamide
ZNS Latex Reagent
2Package1 × 20 mL
1 × 8 mL

References ＊＊1)
Package Insert of Excegran Tablets 100mg andExcegran Powder 20%.
2) The Japanese Society of Therapeutic Drug
Monitoring. [Guideline for therapeutic drug
monitoring (TDM) for antiepileptic drug 2018].
Tokyo: Kanehara Shuppan; 2018. pp. 16-17.
3) In house data, SEKISUI MEDICAL CO., LTD.
Contact
SEKISUI MEDICAL CO., LTD.
international@sekisui.com

ManufacturerSEKISUI 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: 15 months from the date of manufacture(The expiration date is printed on the outerpackage.)3