Analisis Vitamin 9
Transcript of Analisis Vitamin 9
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By: Zaidiyah S.TP, MSc
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Vitamin di definisikan sebagai senyawa
yang mempunyai BM rendah sehingga
manusia yang tergantung senyawa organik
sebagai sumber nutrient membutuhkansenyawa ini dalam jumlah yang sedikit.
Walaupun kebutuhan vitamin sedikit yang
diperoleh dari pola diet/pola makan sehari-hari ketiadaanya/kekurangannya dapat
menyebabkan kelainan fungsi sistem tubuh
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Analisis vitamin dapat dikelompokkan menjadi 3
yaitu;
1. Dengan melibatkan makhluk hidup dan
hewan.2. Dengan menggunakan mikroorganisme
seperti bakteri dan jamur
3. Analisis fisikokimia menggunakan
spectrophotometric, fluorometric,chromatographic, enzymatic, immunological,
and radiometric methods.
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Presisi dan akurasiFaktor ekonomiJumlah sample
Karakteristik sample yang akan dianalisis
Karena sifat vitamin yang sangat sensitivterhadap cahaya, oxigen, pH dan panas
maka dilakukan beberapa tindakanpencegahan untuk menghindari kerusakanvitamin tersebut selama masa analisis
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Dalam beberapa kasus analisis vitamin,
ekstraksik dilakukan pada bagian
biologis samplenya, meliputi beberapa
tindakan berikut ini yaitu; panas, asam,
alkali, pelarut dan enzim yang
digunakan. Pada umumnya prosedur
ekstraksi tergantung jenis vitaminsehingga jenis vitamin yang akan
dianalisis tetap stabil.
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Asam askorbat : Ekstraksi dingin denganasam metaphospone/asam asetat
Vitamin B1 dan B2 : dipanaskan atauautoklav dalam asam atau menggunakanenzim
Niasin : autoklav dalam enzim atau asamVitamin A, E atau D : Ekstraksi pelarut
organik, saponifikasi/penyabunan, dan
reektraksi dengan pelarut organik. Biasanyaditambahkan antioksidan agar vitaminnyatetap stabil dan menghambat prosesoksidasi
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Vitamin A cukup sensitif terhadap
uv/cahaya, udara, temperature tinggi dan
kelembaban. Oleh karena itu terdapat
beberapa langkah untuk menghindari
beberapa pengaruh seperti menghindari
suhu tinggi juga penambahan
antioksidan pada awal prosedur.
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Approximately 0.5 g of freeze-dried sweet potato root wasweighted and put into a centrifugal glass vessel.
5 ml of methanol was gift to the sample and mixed 1
minute on Vortex.
The vessel set was put on the shaker and let it shake 10minute, afterward centrifuge (Sorvall RC-5B Refrigerated
Superspeed Centrifuge serial 820) it at 3500 rpm for 15
minute long.
Then, the supernatant of the samples (clear fraction) was
transferred into 25 ml glass flask. This process was known sample extraction. This process
was repeated for 3-4 times from one sample and brought
together all supernatant of one sample into one flask
(overall 4 extractions of 1 sample/1 flask).
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Furthermore, the flask was filled up to themarked line with methanol. By usingspectrophotometer (Hewlett Packard S 453), thisextract was ready to measure the absorbance (A)
at three different wavelengths as; 470, 653 and666 nm respectively. Methanol was used as blank. Equation to determine concentration
(concentration in solution in g/ml) of
chlorophyll a (Ca) and b (Cb) as well as totalcarotenoids (Cc):Ca = (15.65 x A666)(7.34 x A653)Cb = (27.05 x A653)(11.21 x A666)
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This measurement was determined by titration withsolution 2,6-dichlorophenol-indophenol (DIP)
0.21 g of DIP-Na put in 100 ml of pure water, mixedand filtrated into 1 liter volumetric flask.
Filter was washed several times and filled up theflask to the mark. DIP solution then filled into burette.
5 ml of guava syrup were prepared and immersed in20 ml 5% metaphosphoric acid.
The solution was mixed together until 2 minutes.After homogenizing, the homogenate filled with
distilled water to a volume of 50 ml, mixed andfiltrated.
Ten ml filtrate put in Erlenmeyer flask and titratedwith 2,6 DIP until the color change to light pink
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The content of ascorbic acid can calculated according to formulabelow:Content (mg/100ml guava syrup) =(V (ml) x F1 x (100 ml/SaVo ml) x (ExtrVol/TitrVol)Which:V (Used DIP-dye solution in ml)
F1( Ascorbic acid equivalent of dye solutionexpressed as mg/ml of dye determined before with
standardized ascorbic acid solution (1 mg/1 ml) (three times)).Sa Vol (Sample volume (ml) = 5 ml)ExtrVol (Extraction volume (ml) = 50 ml)Titr Vol ( Aliquot volume (ml) used for titration = 10 ml)
The factor (F1) is the titration volume of 1 ml from 0.1% standardsolution of ascorbic acid in a mixed solution of 1 ml 5%metaphosporic acid and 9 ml distilled water. Ten of that solution istitrated using DIP.
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