Copper
A stock solution containing 1000ppm of copper ions was
prepared from 3.80g of copper nitrate dissolved in 250cm3
distilled water. From the stock solution, standard solutions
of concentrations 1.0ppm, 2.0ppm, 4.0ppm, 6.0ppm,
8.0ppm, 10.0ppm and 12.0ppm were made.
Determination of Thiamine
The method used to determined thiamine was described by
Chopra and Kanwar
[11].
2g of the sample was weighed.
75cm3
of 0.2N HCl was added; this was boiled for 30mins
on a water bath. It was cooled and 5mls of enzymes
solution was added and incubated at 37°C overnight. It was
placed in 100cm3
flask and made to volume with water. It
was filtered and the filtrate was purified by passing it
through silicate column. 5cm3
of acidic KCl was used as
eluate in a conical flask with addition of 3cm3
of alkaline
ferricyanide solution; 15cm3
of isobutanol was added and
shaken for 2 minutes. It was allowed to separate and the
alcohol layer was taken. 2g of anhydrous Na2SO4 was
added to the isobutanol extract. 5cm3
of thiamine solution
was poured in a 50cm3
stoppered flask. Oxidation and
extraction of thiochrome was also carried out as above.
Preparation of sample and standard blank was done taking
5cm3
each 3cm3
of 15% NaOH was used.
UV… Vis was used instead of flourimeter and it was read at
550nm
Caculation
Thiamine content = x/y x 1/5 x 25/v x 1/wt of sample.
Copper
A stock solution containing 1000ppm of copper ions was
prepared from 3.80g of copper nitrate dissolved in 250cm3
distilled water. From the stock solution, standard solutions
of concentrations 1.0ppm, 2.0ppm, 4.0ppm, 6.0ppm,
8.0ppm, 10.0ppm and 12.0ppm were made.
Determination of Thiamine
The method used to determined thiamine was described by
Chopra and Kanwar
[11].
2g of the sample was weighed.
75cm3
of 0.2N HCl was added; this was boiled for 30mins
on a water bath. It was cooled and 5mls of enzymes
solution was added and incubated at 37°C overnight. It was
placed in 100cm3
flask and made to volume with water. It
was filtered and the filtrate was purified by passing it
through silicate column. 5cm3
of acidic KCl was used as
สารละลาย ( eluate ) ในขวดรูปกรวยด้วยการเพิ่ม 3cm3
ของสารละลายเฟอร์ริกไซนาไนด์ด่าง
; 15cm3 ของไอโซบิวทานอล ได้เพิ่มและ
เขย่าเป็นเวลา 2 นาที มันได้รับอนุญาตให้แยก
แอลกอฮอล์ชั้นถูกจับตัวไป 2G ของรัส na2so4 คือ
เพิ่มให้กับไอโซบิวทานอลสารสกัด 5cm3
โซลูชั่นของไนอาซินถูกเทใน 50cm3
stoppered ขวด ออกซิเดชันและ
extraction of thiochrome was also carried out as above.
Preparation of sample and standard blank was done taking
5cm3
each 3cm3
of 15% NaOH was used.
UV… Vis was used instead of flourimeter and it was read at
550nm
Caculation
Thiamine content = x/y x 1/5 x 25/v x 1/wt of sample.
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