Semen Fructose Estimation in Bulls, Pigs, and Man by Dr K Chaudhry

Semen Fructose in Fertility Clinics - Digital book Semen fructose in Bulls and Man - Review Article on Researchgate Hospitals Around The World
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Semen Fructose Estimation in Fertility Clinics For Bulls, Pigs, and Men
By Dr K Chaudhry
FIRST Author of Jaypee Brothers
Developer & Maker of Stable Indole 3-Acetic acid Reagent

HISTORY :- As Biochemistry Reagent Manufacturer, I found most reagents deteriorating by oxidation. I stabilized many reagents with Thiourea and other antioxidants. I used Tap water in most colorimetric reagents. With time I began using, Thiorea and Benzoic acid as default additives in Tap water used for Colorimetric reagent making.
Around year 1999, with my intuitive mind, I encountered Indole 3-acetic acid Reagent stable for just a few hours. I worked overtime on Chemistry of Indole 3-acetic acid. I made Indole 3-acetic acid stable at Room Temperature. Dr Subhash Bhargava had been my Ideal and Guide. He tried the reagent and commented as PERFECT. The reagent was soon picked for regular use by Dr Rakesh Sharma, Dr Madhu Gupta, Dr Madhu Dua, Dr Jitender Bhatia; later by Dr Sandeep Dogra (Jammu), Dr Pran Nath Seth.
During 2019, on Researchgate a scientist mentioned CASA as Standard equipment for Fertility assessment of Semen. I may be wrong but CASA is perhaps not used in any Medical College in India. CASA testing is available at some commercial laboratories in Delhi, Hyderabad, Indore, may be less than a dozen more. Semen Fructose Drop Test is much more specific than CASA Testing with multiple parameters. Also Semen Fructore is Time sensitive. Transportation time may be strong deterrant for CASA Testing. Reproduction Scientists may suggest a laboratory around to receive Free Trial Kit. Entire reagent needs would be supplied Free for Research projects. Some reputed Limited companies marketed kits from my OEM packs. Facing too many issues from Government in consumer sale within India and overseas, I would concentrate on OEM Supplies worldwide. Recommendations from Reproduction Scientists solicited.

Semen Fructose Estimation
For colorimetric determination of fructose in semen

Introduction
Fructose in presence of hydrochloric acid, forms a pink coloured complex with indole-3-acetic acid. The complex has maximum absorbance at 500-530 nm. The product is now open to public use. Laboratories may prepare and use reagents. Diagnostic companies may manufacture and distribute; no royalty payment expected.

SOURCE : https://www.researchgate.net/post/Semen_Fructose_Estimation_in_Animals_and_Man-Infinitely_Stable_Indole_3-acetic_acid_reagent

Semen Fructose Estimation in Animals and Man - Infinitely Stable Indole 3-acetic acid reagent

I am grateful to friends across the globe for patronage to Semen Fructose Estimation Kit developed by me and manufactured by my family enterprise Maa Vaishno diagnostics. Perhaps marketing and procurement problems have been a bottleneck in use of this method of immense importance in cattle and human infertility.

On National Republic Day of year 2020, I am pleased to declare my Reagent Formulation open to public use.

Diagnostic laboratories may prepare and use my Reagent.

Diagnostic companies may manufacture and distribute; no royalty expected.

Method described below and also viewable at http://indianmedical.net/fructose

Reagents 1. DoctorKC' Fructose Reagent - Stable Indole 3-acetic acid Reagent. The reagent is infinitely stable at room temperature.

2. Hydrochloric acid LR

3. Standard : 200 mg/dl 5 ml

Procedure

For Semen Fructose Drop Test in evaluation of Sperm Activity, Standard reading is not requird.

1. Pour over 0.005 ml sample, 5 ml Hydrochloric acid. 2. Add 0.1 ml Fructose Reagent. Mix by inversion. 3. Incubate at 50ºC for exact 50 minutes. 4. Cool and read absorbance of Sample and Standard against Blank adjusted to zero with bluish green filter (500-530 nm).

For standard, take 0.005 ml standard in place of sample in step 1. For blank, omit sample/standard addition. Proceed further with both tubes as with the test.

For Semen Fructose Drop Test, use Sample reading of 30 minutes old sample (Abs30) and 240 minutes old sample (Abs240).

Calculation
                              Abs Sample
Fructose = ------------------------------- x 200 mg/dl
                            Abs Standard

Fructose Drop using Standard

                                Fructose at 30 minutes - Fructose at 240 minutes
Fructose Drop = -------------------------------------------------------------------- x 100 percent
                                                   Fructose at 30 minutes

Fructose Drop without Standard

                          Abs30 - Abs240
Fructose drop = ----------------------------- x 100 percent
                                        Abs30

Diagnostic significance

1. Fructose level in normal semen is over 200 mg/dl at 30 minutes after ejaculation. Fructose is utilised rapidly during sperm activity and fructose level drops by over 40% at 4 hours after ejaculation.

In FRUCTOSE DROP TEST, fructose level is determined at 30 minutes and at 4 hours after ejaculation. The result is reported as % drop from the first level. At least 40% drop over 210 minutes indicates Normal Sperm activity. Semen Quality evaluation is based upon percent drop over 210 (30, 240) minutes. Absolute semen fructose value has little role. Hence to need to standardize the Kit. If Semen Fructose Drop results do not match Computer Assisted Semen Analysis (CASA) analysis, the Semen Fructose Drop would be construed as Reference method being more directly related to Sperm activity.

Fructose level in semen with dead sperms is 150-250 mg/dl and may show a little drop over time.

2. Absence of both fructose and spermatozoa in the semen indicates obstruction distal to the ejaculatory duct. These findings may be used to confirm vasectomy operation.

3 Reappearance of fructose in semen is used to assess the success of recanalization operation.

Other uses
1. Urine fructose estimation.
2. Urine inulin estimation in inulin tolerance test. Inulin is hydrolysed into glucose and fructose in the procedure itself.
3. Fructose estimation in sugar industry. Here also sucrose is hydrolysed into glucose and fructose in the procedure itself.