Histamine quantitative test kit for seafood 61341 | Kikkoman

Product detail

The histamine content of tuna and of canned products containing histamine is determined by various methods such as HPLC, EIA and enzymatic colorimetric methods. Among these, the enzymatic colorimetric method is regarded as simple, fast to analyse and as reliable as the conventional methods.

1. Features of the histamine quantitative test kit for seafood

– Simple and safe: only the reagent has to be dissolved and mixed with the sample, and the high specificity of the enzyme means no hazardous chemicals such as strong acids or organic solvents are needed. Easy extraction: no clean-up is required, whereas the HPLC method needs clean-up first to remove interfering substances. Easy to use: no complex procedure such as HPLC or EIA is required.

– Fast: the reaction takes only 15 minutes, and for live fish samples the test takes about 1 hour from sample preparation to result. Third, it is accurate, because AOAC (PTM) certification has been obtained and the tests have been shown to correlate highly with other reference methods

2. Components of the histamine test kit

– Enzyme reagent × 6 bottles with a blue label

– Colorimetric reagent × 6 bottles with a red label

– Buffer solution × 3 bottles with a pink label

– Histamine standard (30 mL) × 1 bottle with a blue label

3. Equipment, consumables and instruments to prepare

– Sample grinder/blender (this instrument is not needed for fish sauce samples)

– Technical balance (able to weigh a 1 g sample)

– Heat-resistant plastic tubes with caps (50 ml heat-resistant centrifuge tubes with caps), rack

– Micropipette 100-1000 µl and blue 100-1000 µl tips, tip box (if available)

– Small chemical spatula

– Double-distilled water

– Filter paper No. 5C, filter funnel

– 15 ml centrifuge tubes, 50 ml centrifuge tubes, centrifuge tube rack

– Water bath (or a stove and a pot)

– 0.1 M EDTA 2Na solution, pH 8 / 0.5 M EDTA solution, pH 8

– DPM2-ABS spectrophotometer (Kyoritsu)

– Vortex mixer

– Medical gloves and heat-resistant gloves (if available)

– 50 ml measuring cylinder

– Stopwatch

* Note:

– Histamine is very readily adsorbed onto glass. Plastic tubes or similar should be used.

– This kit is a histamine test for live fish. However, by changing the sample treatment method it can be applied to other samples.

4. Test procedure of the Kikkoman histamine test kit

4.1. Reagents

– Distilled or deionised water

– 0.1 M EDTA 2Na 2H2O (pH 8): sample treatment buffer (A 0.5 M EDTA solution (pH 8) can be used as the sample treatment buffer by diluting it 5-fold).

– Dissolve 37.2 g EDTA 2Na2H2O in water to make 750 mL. Adjust the pH to 8.0 with potassium hydroxide solution. Make this solution up to 1 L with distilled water.

4.2. Instructions

1. Prepare the sample solution

– Put 250 µl fish sauce into a 50 ml test tube and dilute with the sample treatment buffer to 50 ml, then mix well with a vortex mixer. *200-fold dilution

– Filter the solution containing the histamine extract through filter paper (No. 5C).

– Take 1 ml or more and mix well. The sample is ready.

2. Prepare the reagents

– Dissolve the colorimetric reagent (red label) in 11 ml distilled water.

– Dissolve the enzyme reagent (blue label) in 6 ml buffer solution (pink label).

3. Measurement

(1) Prepare 4 test tubes and add the reagents according to the table below. Repeat 3 times for one sample

（Sample, sample blank, standard solution, reagent blank）×3＝12 tubes

	(1)Sample Es	(2) Sample Blank Eb	(3)Standard Solution Estd	(4) Reagent blank Ec
Water (mL)	－	－	－	0.5
Buffer (mL)	－	0.5	－	0.5
Sample solution (mL)	0.5	0.5	－	－
Histamine standard solution (mL)	－	－	0.5	－
Colorimetric solution (mL)	0.5	0.5	0.5	0.5
Enzyme solution (mL)	0.5	－	0.5	－