Chemical tests to evaluate semen
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CHEMICAL TESTS TO EVALUATE SEMEN
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Chemical tests available are
DNA estimation
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The available literature shows that the quantity and quality of DNA content of sperm alters during storage, seasonal variation and aging of the male.
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This alteration of DNA content may be co-related with fertility.
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Gledhill (1966) reported that in infertile bulls the amount of DNA content may not vary but the quality of basic protein alters.
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Ascorbic acid content
Procedure
- This method was described by Roy et al., 1950.
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Take 0.2 ml of fresh semen in a sterile test tube.
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Add 0.8 ml trichlor acetic acid.
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Mix the contents well.
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Filter the mixture through Whatman filter paper No. 1.
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Add 0.05 ml of 2,6 dichlorophenol indophenol solution in a centrifuge tube.
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Titer it against protein free filtrate of semen in a micropipette.
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Check the end point where the pink color disappears.
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A blank solution of 0.2 ml of standard ascorbic acid plus 0.8 ml of 10% trichlor acetic acid is titrated against 0.05 ml of 2, 6 dichlorophenol indophenol dye.
Titration reading of standard
Ascorbic acid concentration = ------------------------------------ x 1.2
Titration reading of unknown
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Preparation of dye
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Extract 100 mg of 2, 6 dichlorophenol indophenol through a filter paper with 25 ml of boiling distilled water. It can be kept as stock solution. This can be stored in refrigerated temperature. On the day of estimation 5 ml of stock solution is made up to 100 ml by adding freshly boiled and cooled distilled water.
Standard ascorbic acid
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Stock solution can be prepared by dissolving 60 mg of ascorbic acid in 5% acetic acid and the volume 100 ml. For routine use 1 ml of the the stock solution is diluted with 5% actetic acid to 100 ml. Thus 1.2 gm of ascorbic acid are present in 100 ml of solution.
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Last modified: Monday, 4 June 2012, 10:08 AM
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