Judul: Identification of Pork DNA Fragments in Gelatin Samples Using Real Time Polymerase Chain Reaction (qPCR) methods
Abstrak: Gelatin merupakan suatu jenis protein yang diperoleh melalui hidrolisis parsial kolagen dari kulit dan tulang hewan yang memiliki banyak fungsi untuk keperluan berbagai industri. Kebutuhan gelatin dalam negeri umumnya dipenuhi secara impor. Gelatin yang berasal dari kulit babi adalah yang tertinggi dari total produksi gelatin di dunia. Produk babi dan turunannya termasuk gelatin babi dilarang untuk dikonsumsi oleh Muslim, sehingga diperlukan adanya pengujian untuk mengetahui sumber gelatin. Penelitian ini bertujuan untuk mengidentifikasi fragmen DNA babi pada sampel gelatin dengan menggunakan metode real time Polymerase Chain Reaction (qPCR) dengan primer dan probe sudah termasuk ke dalam master mix mericon Pig Kit. Sebanyak 53 sampel gelatin diekstraksi sesuai dengan prosedur standar dari DNeasy mericon Food Manual Book. Hasil ekstraksi sampel gelatin dikuantifikasi menggunakan nanodrop spectrofotometer 2000c dan diperiksa menggunakan kit komersial. Proses amplifikasi dilakukan sesuai dengan prosedur standar dari DNeasy mericon Pig Kit Handbook. Hasil uji qPCR mampu mengidentifikasi adanya fragmen DNA babi pada sampel gelatin yang ditunjukkan dengan nilai Ct<45. Sebanyak 4 sampel gelatin dari 53 sampel positif mengandung DNA babi, sedangkan 49 sampel gelatin menunjukkan hasil negatif. Uji qPCR dapat dijadikan sebagai metode pengujian untuk mengidentifikasi adanya fragmen DNA babi pada sampel gelatin., Gelatin is a protein obtained by hydrolysis of collagen from animal skin and bone that has great importance in many industry. Domestic gelatin is generally obtained by import. The raw materials used in gelatin production are highly from porcine sources. Porcine gelatin is forbidden for Muslim. For this reason, analytical methods to detect gelatin are needed to make sure the source of gelatin. The aim of this research was to identify the presence of pork DNA fragments in gelatin by real time PCR (qPCR) methods. The primer and probes of qPCR were included in mericon Pig Kit master mix. The samples consisted of 53 gelatins. They were extrated according to the procedure of DNeasy mericon Food Manual Book. The result of extracted gelatins was quantified by nanodrop spectrofotometer 2000c and examined by a commercial kit. The amplification process of gelatins was carried out according to the procedure of DNeasy mericon Pig Kit Handbook. The result showed that gelatins with pork DNA can be identified by Ct value<45. There were 4 samples positive contained pork DNA, while 49 samples were negatives. The qPCR methods can be used to identify the presence of pork DNA fragments in gelatin samples.
Keyword: DNA, gelatin, pork, qPCR

Judul: Sensitivitas Metode Real-Time Polymerase Chain Reaction dalam Mendeteksi DNA Gelatin Babi Pada Produk Confectionery
Abstrak: Gelatin merupakan salah satu ingredien yang banyak digunakan sebagai bahan pembentuk gel dan pengental dalam produk pangan. Tulang dan kulit babi berpotensi untuk digunakan sebagai bahan baku dalam produksi gelatin komersial. Gelatin dikategorikan sebagai bahan kritis dari segi kehalalannya. Real-Time Polymerase Chain Reaction (real-time PCR) banyak digunakan untuk mendeteksi residu DNA babi di dalam produk pangan. Hasil penelitian sebelumnya menunjukkan bahwa metode real-time PCR dengan primer Tanabe dan Internal Positive Control (IPC) memiliki parameter validasi yang baik seperti robustness, linieritas, sensitivitas atau limit of detection (LOD) konsentrasi DNA babi, spesifisitas dan efesiensi amplifikasi. Namun belum diketahui LOD untuk mendeteksi DNA babi pada produk confectionery berbahan baku campuran gelatin babi dan gelatin sapi pada berbagai rasio. Penelitian diawali dengan penentuan LOD (% b/b) gelatin babi di dalam campuran gelatin sapi dan gelatin babi. Hasil menunjukkan bahwa nilai LOD pada sampel tersebut adalah 0,01 % b/b dengan nilai Ct 39,20±1,718. Tahap kedua adalah menentukan LOD (% b/b) gelatin babi di dalam produk confectionery (lozenges, gummy candy, dan marshmallow). LOD (% b/b) gelatin babi pada lozenges dan gummy candy adalah 0,1 % b/b dengan nilai Ct masing-masing sebesar 40,93±0,15 dan 38,72±0,18, sedangkan marshmallow adalah 0,01 % b/b dengan nilai Ct 41,14±2,96. Tahap terakhir adalah mengaplikasikan metode tersebut terhadap produk confectionery komersial yang diketahui mengandung gelatin babi. Hasil menunjukkan semua sampel dapat terdeteksi dengan baik. Hasil penelitian ini menunjukkan bahwa metode real-time PCR dengan primer Tanabe dan IPC mampu mendeteksi DNA babi pada produk confectionery berbahan baku gelatin dengan sensitivitas yang tinggi., Gelatine is a widely used gelling and thickening agent in confectionery products. Commercially, pig's bone and skin are commonly employed as raw materials for gelatine production. Due to its source, gelatine's halal status is critical. Real-time Polymerase Chain Reaction (real-time PCR) is a commonly used analytical tool to detect porcine residue in food products. The real-time PCR method, using the Tanabe primer and Internal Positive Control (IPC), has robust validation parameters, including linearity, robustness, LOD of porcine DNA, specificity, and amplification efficiency. However, the sensitivity or limit of detection (LOD) of porcine DNA in confectionery products made from mixtures of porcine and bovine gelatin at varying ratios has not been determined. This study aimed to determine the LOD (% w/w) of porcine gelatine in bovine gelatine, followed by determining the LOD (% w/w) of porcine gelatine in various confectionery products, such as lozenges, gummy candy, and marshmallows. The results revealed an LOD (% w/w) of 0.01% with a Ct value of 39.20±1.718 for porcine gelatine in bovine gelatine. The LOD (% w/w) for porcine gelatine in lozenges and gummy candy was 0.1% with Ct values of 40.93±0.15 and 38.72±0.18, respectively. The marshmallow LOD was 0.01% w/w with a Ct value of 41.14±2.96. Finally, this method was successfully applied to commercial confectionery products containing porcine gelatin, detecting its presence with high sensitivity.
Keyword: confectionery product, LOD, Porcine gelatin, real-time PCR

Judul: Thermal Characteristics and Dynamics of Tropical Cyclone over Western North Pacific Ocean: A Case Study of Typhoon Songda (Chedeng) 2011
Abstrak: The frequent occasion of Tropical Cyclone (TC) over Western North Pacific (WNP) Ocean lies from 10˚N to 26˚N and 121˚E to 170˚E. Sources of TC development come from warm ocean temperature where latent heat is released during condensation processes and subsequently concentrated in the boundary layer. Relative humidity at 850-700 hPa is responsible to produce the most intense storm. Meanwhile, weak vertical wind shear between 850-200 hPa is one of the dynamical factors that lead to the development of TC. Pressure level of 850-200 hPa is chosen to minimize the thermodynamic effect in the lower troposphere. Typhoon Songda (Chedeng in the Philippines) or TSC lasted from May 18th to 30th, 2011 and strengthened into powerful Super Typhoon stage between May 24th and 27th. Long curving track and the exhibition of all sequences of TC life cycle, TSC is then considered as an ideal case of TC. The objective of the study with case of TSC is to investigate the role of thermal characteristics and dynamics on TC intensity. The research is carried out by using Advanced Research of Weather Research and Forecasting model (WRF ARW model) version 3.3. Two domains are employed in the model. The first domain situate from 02˚N to 40˚N and 118˚E to 149˚E and cover spatial resolution () of 33.1 km. The second domain situate from 09˚N to 27˚N and 119˚E to 132˚E with of 11 km. Timestep of the model () is 120 seconds. The model is supported by the following physical parameterization, i.e., Yonsei University Scheme (YSU) in the boundary layer, Kain-Fritsch scheme (KF) in the cumulus, and WRF Single-Moment 3-class scheme in the microphysics. Initial meteorological condition prior to the formation of the cyclone is generated using WRF ARW standard-release model. Sensitivity of the model output is obtained from a selected single-parameter change scenarios, namely, 10 % to 20 % decrease and increase of wind shear initial data at 850-200 hPa, and by 10 % to 20 % decrease of relative humidity initial data at 850-700 hPa. The result showed that the negative correlations between wind shear and intensity are found in wind shear scenarios. Wind shears of 10 ms-1 which lead to strengthening of the wind intensity in the early period of Typhoon development are found in wind shear scenarios. The effect of wind shear changes on wind intensity is statistically significant in the 20 % wind shear scenarios. During mature stage, weak wind intensity occurred when wind shear increase as given in the WRF standard-release and the 20 % decreasing scenario. Different situation is found in the 20 % increasing scenario where wind shear tend to weak and lead to strong wind intensity at this stage. Meanwhile, the reduction of moisture supply at 850-700 hPa resulting from 20 % decreasing relative humidity play a major role in the weakening of wind intensity and reduces mechanical energy of the cycle process as much as 300 J kg-1.
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