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albumins were separated and placed in separate petri dishes 3) In the ammonia receiving flask, 10mL of boric acid
of known weight. (4%) was added with three drops mixture of methyl red
The percentage of moisture content was calculated and Bromocresol Green dye. Collecting up to 25-30 mL
according to the following formula: in a receiving conical flask after the end of the
Moisture content % = [(g moist sample - g dry sample) distillation process.
/ g moist sample]×100 4) Titration: the collected solution in the receiving conical
Also, the total solids of samples were calculated flask was titrated with 0.1M of HCL, and the titre was
according to the following equation: recorded from which the amount of nitrogen content was
Total solids % = 100 – Moisture content % measured according to this equation:
*After the determination of moisture, the protein % Nitrogen= [{mL (titre–B)×M HCl×dilution
content was determined in all the samples using Kjeldahl factor×14.007}/(mg sample×10)]×100 Eq.1
procedure. Where:
Determination of protein by Kjeldahl method M HCL= Molarity of hydrochloric acid, B= Blank
Reagents preparation: To convert dry weight to wet weight, the moisture
1. Sodium hydroxide solution, 40%: 200g of sodium content of the raw material (i.e., wet weight ) must be found.
hydroxide was weighed and dissolved in distilled water. The conversion formula is:
The volume was made up to 500mL in a volumetric flask. Dry weight (D.Wt) = wet weight (W.Wt)×[(100-
2. Boric acid solution 4%: Weighing 20g of Boric acid moisture percentage)/100] Eq.2
and dissolved in distilled water, making up the volume to To convert from dry weight to wet weight:
500mL in a volumetric flask. Wet weight (W.Wt)= dry weight (D.Wt)×[100/(100-
3. Methyl red / bromocresol green indicator moisure percentage)] Eq.3
solution: Dissolving 100mg methyl red and diluting to This formula is used to find protein % in the samples:
100mL in 95% ethanol. Followed by dissolving 100mg % P = % N × CF Eq.4
Bromocresol green and diluted to 100mL in 95% ethanol. Where:
Finally mixing one part of the methyl red solution with % P = Protein , % N = Nitrogen , CF = Conversion Factor
five parts of the Bromocresol green solution. Statistical analysis
4. Blank: Contains all the reagents used except for the Data collected for all parameters were analyzed by a
sample in every batch, so to subtract reagents effects from completely randomized design (CRD). The difference
the sample nitrogen (Puwastien et al., 2011). among mean was tested by Duncan's multiple range tests,
Procedure according to P ≤ 0.01 significance, and the result of
Protein total nitrogen content was determined in 1 statistical analysis results are shown as mean value and
gm of the dried samples using micro- Kjeldahl technique. standard error in tables. The statistical calculations of the
The Kjeldahl method was performed according to method results were completed using the XLSTAT (2016) program.
981.10 of the AOAC International (Latimer, 2016); RESULTS AND DISCUSSION
(William, 2000, Persson, 2008, Puwastien et al., 2011). The
three steps of the Kjeldahl method were carefully carried Results of determination of protein, moisture in animal
out in sequence as follows: products
1) Digestion: weighing about 1g in triplicate of the dried The results obtained in this study are the
samples (red meat, chicken, Beef hot dog and egg) , into determination of moisture and protein in some animal
the Kjeldahl flask, followed by the addition of 30mL of products using Kjeldahl procedure (Table 1).
concentrated sulphuric acid (95-97%), 0.4g of copper To each product sample, the mean value of analysis
sulfate, and 3.5g of potassium sulfate. The mixture was and standard error of the mean were measured. Sample
heated in a fume cupboard slowly to prevent excessive constituent values were ranged from 11.59 ( Egg-Albumin )
frothing; then, the digestion was continued at 400°C for to 22.01% (Fresh Chicken breast) for protein, containing
2.5-3 hrs until the color of the mixture changed to 50.0 ( Egg-yolk ) to 87.6 ( Egg-albumin ) of moisture. From
iridescent blue color. The solution was left to cool down the results showen, it can be concluded that the standard
and diluted with distilled water to 100mL. error of the mean value is the highest (0.47) in frozen
2) Distillation:10 ml of the digested solution was carefully silverside (veal meat), while in beef hot dog recorded the
added to 10 ml NaOH (40%) and fixed to the distillation lowest value (0.07).
device.
Table 1. Percentages of moisture and protein contents in some animal products.
%Protein %Moisture
Sample Type
%Mean Standards or declared value SEM %Mean SEM
Fresh, Breast 22.01 24.00 0.17 76.5 0.17
Chicken meat
Fresh, Thigh 19.74 20.90 0.08 78.57 0.33
Fresh, Silverside 20.55 0.33 76.8 0.09
Veal meat 22.70
Frozen, Silverside 19.08 0.47 76.5 0.10
Egg-Albumin 11.59 12.00 0.10 87.6 0.25
Egg
Egg-Yolk 14.73 16.10 0.29 50.0 0.55
Beef meat Hot dog 12.88 13.00 0.07 73.9 0.17
*CV-Coefficient of variation = (standard deviation ÷ mean)*100
*VI- variation interval (span) = max. value – min. value
*SEM-Standard error of the mean
344
J. of Food and Dairy Sci., Mansoura Univ., Vol 11 (12), December, 2020
345
Sazan Dana Jamal et al.
تقدير البروتين في بعض المنتجات الحيوانية من أسواق السليمانية باستخدام طريقة كلدال
2 و زيد خلف خضر1 دارا محمد جميل، * 1 سازان دانا جمال
. العراق، اقليم كردستان، جامعة السليمانية، كلية علوم الهندسة الزراعية، قسم علوم االغذية ومراقبة الجودة1
. العراق، اقليم كردستان، جامعة السليمانية، كلية علوم الهندسة الزراعية، قسم علوم الحيوان2
يعد قياس إجمالي. تم قياس البروتين الخام لعينات المنتجات الحيوانية من خالل تحديد محتوى النيتروجين الكلي باستخدام طريقة كلدال، في هذه الدراسة
النيتروجين بواسطة كلدال هو األسلوب المرجعي العالمي لتقدير محتوى البروتين في المنتجات الحيوانية والنباتية ويستخدم في كل من المعايرة والتحقق من صحة
بنا ًء على. مصادر البروتين الحيواني التي تم تحليلها في هذه الدراسة كانت عينات من لحم العجل ولحم الدجاج والبيض والنقانق.الطرق البديلة في تقدير البروتين
صدور الدجاج > ظهر الفخذ الطازج > فخذ الدجاج الطازج > ظهر: كان تسلسل محتوى البروتين في العينات هو، النتائج التى تم الوصول اليها في هذا البحث
اما نتائج بياض، وتطابقت نسبة البروتين مع النسبة المثبتة على علبة نقانق اللحم البقري.الفخذة المجمدة > صفار البيض > نقانق اللحم بقري > بياض البيض
. لم تظهر فروق معنوية مقارنة بالحدود القياسية،البيض و فخذ الدجاج الطازج
346