3.4.1.2 Reagents

Solution A. Pararosaline was prepared by dissolving 1 g powder in 20 ml double

2.Solution B. Sodium nitrate (4%) was prepared by dissolving 400 mg in 10 ml

3.Solution C. Alpha naphthyl acetate was prepared by 50 mg powder in 2.5 ml

5.Harris haematoxylin. (500 ml) was prepared by dissolving 2.5 g powder in 50 ml ethyl glycol added to a day before prepared supersaturated solution of alu (90 g in 500 ml double distilled water) and sodium iodide or potassium iodide (20 mg). The stain was stirred overnight, filtered and stored in a dark

Bone marrow from both femurs of mice was collected in PBS, washed thrice and smeared over the slides. Air dried slides were fixed in freshly prepared fixative 30 s at 4°C and dipped in double distilled water thrice. Air dried slides were incubated at room temperature in the following freshly prepared filtered solution. 1.2 ml solution A and 1.2 ml solution B was mixed well and allowed to react for 1 min after which solution C was added and was made up to 50 ml solution by

Slides were incubated in above solution for 45 min at 37°C. After incubation slides were washed in double distilled water for 10 min and counter stained with haematoxylin for 1 min. After staining slides were washed in water for long time and observed under microscope (100�, oil immersion) for scoring positive and

3.4 Determination of the effect of Xancus pyrum on enzyme levels in cisplatin

Liver homogenates were made in ice cold Tris buffer (0.1 M pH 7.4) and was used for the estimation of SGOT, SGPT, urea and creatinine. Serum was also used to

Alanine aminotransferase (ALT) catalyses the transamination of L-alanine and α-ketoglutarate to form pyruvate and L-glutamate. Pyruvate so formed is coupled with 2,4-dinitrophenyl hydrazine (2,4-DNPH) to form a corresponding hydrazone,

Pyruvate þ 2, 4 � DNPH ⇄ corresponding hydrazone brown colour ð Þ (2)

L � alanine þ α � ketoglutarate ⇄ pyruvate þ L � glutamate (1)

a brown coloured complex in alkaline medium and this can be measured

distilled water and 5 ml HCl. Gently warmed solution is filtered and stored in

dark at 4°C.

bottle.

3.3.4.3 Procedure

phosphate buffer (pH 7.4).

treated animals

3.4.1.1 Principle

colorimetrically [13].

56

estimate all the above parameters.

negative alpha esterase cells out of 4000 cells.

3.4.1 Estimation of SGPT (Span Diagnostics Ltd., Surat, India)

double distilled water.

Invertebrates - Ecophysiology and Management

glycol monoethyl ether.

4.Phosphate buffer (pH 7.4)


Working reagent preparation:

