Alkaline hematin D تحضير كيتس ال

Alkaline haematin D reagentتحضير ال 

1) Components:

==>  Sodium hydroxide (NaOH) …………… 4 g (Role = Formation of Alkaline Hematin D)
==>  Triton X-100 (or equivalent) ………… 25 g (Role = Detergent to dissolve lipid of the RBCs membrane to rupture it and release Hb)
==>  Distilled water ………………………………. 1000ml (Role = solvent for preparation of the reagent)

طبعا المكونات سهلة جدا ومفهاش اى حاجة محتاجة شرح

2) Function:

First the detergent dissolve the lipids embedded in the RBCs membrane leading to formation of pores in the membrane …… this leads to leaks of hemoglobin from these pores (i.e. hemolysis) ….. we do this, because we can’t measure the hemoglobin concentration while it is contained within the RBCs

Then, NaOH react with the released hemoglobin to form the Alkaline Hematin D-575 which is stable colored compound that can be measured photo-metrically by the spectrophotometer of colorimeter and whose concentration equals the real hemoglobin concentration   

لماذا تدمر المنظفات جدار كرات الدم الحمراء؟؟؟ اضغط هنا

كل اللى بيحصل … ان فالاول الdetergent وده ترجمته بالبلدى منظف وده كل مهمته انه يدوب الليبد او الدهون اللى فغشاء كرة الدم الحمرا عشان ده هيعمل ثقوب فيه وبالتالى هيخرج الهيموجلوبين برة الخلايا عشان طبعا احنا مشهتعرف نقيس الهيموجلوبين وهو جوة الخلايا …. وبعد كدة ال NaOH يتفاعل مع الهيموجلوبين ويكون مركب الalkaline hemtin D-575 وده اللى بنقيس تركيزه بالسبكتروفيتوميتر

3) Preparation:

==> Dissolve the sodium hydroxide in the distilled water in a clean conical flask.

==> Stir using a glass rod until the crystals have completely dissolved.

==> Add the Triton X-100 (or equivalent) and mix well.

Grade 1 filter paper whatman==> Filter the solution into a clean glass-stoppered reagent bottle, using Whatman No. 1 (or equivalent) filter-paper.

==> Label the bottle “ALKALINE HAEMATIN D REAGENT” and write the date.

==> Store at room temperature (20–25°C).

==> Check the quality of the solution (see the quality control below).

التحضير سهل جدا طريقته واللى مشعارف يترجمه ومشمفاهمه يكتبلى كومنت

NOTES:


==> Alkaline haematin D (AHD) reagent will keep for several months at 20–25°C. If a precipitate forms during storage, the reagent should be filtered before use.

==> Use filtered rainwater if distilled water is not available.

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Quality control of alkaline hematin D reagent

An alkaline haematin D standard solution (AHD standard) supplied by the central laboratory is used to test the quality of new batches of AHD reagent in peripheral level laboratories.

1) Fill a clean cuvette with distilled water. Place the cuvette in the cuvette chamber and adjust the haemoglobinometer or colorimeter to read zero at 540nm wavelength.

2) Replace the distilled water with AHD reagent. The haemoglobinometer or colorimeter should read zero.

3) Pipette 20ml of AHD standard into a test-tube containing 3 ml of the freshly prepared AHD reagent (1:150 dilution).

4) Measure the haemoglobin concentration of the AHD standard (اضغط هنا لتشاهد الطريقة).

5) Repeat the procedure using the previous batch of AHD reagent. Compare the results.

6) If the haemoglobin values differ by more than 5 g/l, discard the freshly prepared AHD reagent and prepare a new batch, paying attention to accurate measurement of the constituents and the cleanliness of the glassware.

The AHD standard stock solution will keep for 8 months at 4–8°C.

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