Sintesis dan Karakterisasi Kolagen dari Tulang Bovine Menggunakan Metode Ekstraksi Asam Asetat-Ultrasonik

Amalia, Lailyta Shafira (2026) Sintesis dan Karakterisasi Kolagen dari Tulang Bovine Menggunakan Metode Ekstraksi Asam Asetat-Ultrasonik. Masters thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Kolagen merupakan komponen utama matriks ekstraseluler yang memiliki peran penting dalam rekayasa jaringan dan umum digunakan dalam aplikasi biomaterial, kosmetik, dan farmasi. Salah satu tantangan utama dalam ekstraksi kolagen adalah memperoleh yield dengan integritas struktur molekul yang sesuai dengan standar biomaterial untuk aplikasi medis. Penelitian ini bertujuan untuk mengevaluasi efektivitas metode ekstraksi ultrasound assisted extraction (UAE) dalam pelarut asam asetat terhadap tulang bovine. Metode tersebut dipilih untuk meningkatkan kinetika transfer massa melalui mekanisme kavitasi akustik, sehingga meningkatkan efisiensi ekstraksi. Proses ekstraksi kolagen diawali dengan preparasi tulang bovine, meliputi degreasing dan demineralisasi, untuk eliminasi kontaminan lipid serta mineral anorganik. Matriks tulang yang telah dipreparasi kemudian mengalami proses solubilisasi dalam asam asetat dengan paparan gelombang ultrasonik. Variabel proses seperti anatomi tulang sapi, modifikasi asam, dan durasi ultrasonik dievaluasi untuk menentukan hasil yang optimal. Karakterisasi fisikokimia dan biologis dilakukan meliputi analisis gugus fungsi (FTIR), berat molekul (SDS-PAGE), morfologi permukaan (SEM), sifat termal (DSC-TGA), serta uji biokompatibilitas (MTT Assay). Hasil penelitian menunjukkan bahwa metode ultrasonik secara signifikan meningkatkan perolehan yield kolagen hingga mencapai 11,8% pada durasi optimal 30 menit. Analisis FTIR mengonfirmasi keutuhan gugus fungsional Amida A, B, I, II, dan III yang mengindikasikan struktur triple helix tetap terjaga. Hal ini divalidasi dengan pengujian termal DSC seluruh kolagen memperlihatkan stabilitas termal yang baik dengan suhu denaturasi (Td) tertinggi mencapai 158℃, pengujian SEM memvisualisasikan morfologi serat-serat berpori dan pada kolagen kontrol terdapat gumpalan yang mengindikasikan adanya residu yang tertinggal. Namun, pola pita α dan β pada SDS-PAGE tidak tampak di rentang berat molekul yang seharusnya. Hal tersebut dikarenakan struktur molekul protein sudah terurai menjadi peptida-peptida dengan berat molekul di bawah 25 kDa. Uji sitotoksisitas kolagen sebagian besar menunjukkan viabilitas sel di atas 70%, membuktikan bahwa kolagen hasil ekstraksi bersifat non-toksik dan memenuhi kriteria kolagen standar sebagai bahan baku biomaterial medis.
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Collagen is a key component of the extracellular matrix that has an important role in tissue engineering and is commonly used in biomaterials, cosmetics, and pharmaceutical applications. One of the main challenges in collagen extraction is obtaining yields with molecular structural integrity that conform to biomaterial standards for medical applications. This study aims to evaluate the effectiveness of the ultrasound assisted extraction (UAE) method in acetic acid solvent on bovine bone. The method was chosen to improve the kinetics of mass transfer through the acoustic cavitation mechanism, thereby improving the efficiency of extraction. The collagen extraction process begins with the preparation of bovine bones, including degreasing and demineralization, for the elimination of lipid contaminants and inorganic minerals. The prepared bone matrix then undergoes a solubilization process in acetic acid by exposure to ultrasonic waves. Process variables such as bovine bone anatomy, acid modification, and ultrasonic duration are evaluated to determine optimal results. Physicochemical and biological characterization was carried out including functional group analysis (FTIR), molecular weight (SDS-PAGE), surface morphology (SEM), thermal properties (DSC-TGA), and biocompatibility test (MTT Assay). The results showed that the ultrasonic method significantly increased the acquisition of collagen yield up to 11.8% at the optimal duration of 30 minutes. FTIR analysis confirmed the integrity of the functional groups of Amida A, B, I, II, and III indicating that the triple helix structure was maintained. This was validated by the DSC thermal test of the whole collagen showing good thermal stability with the highest denaturation temperature (Td) reaching 158°C, the SEM test visualized the morphology of the porous fibers and in the control collagen, there were aggregates indicating the presence of residues left behind. However, the band pattern α and β on SDS-PAGE is not visible in the supposed molecular weight range. This is because the molecular structure of proteins has broken down into peptides with a molecular weight of less than 25 kDa. Collagen cytotoxicity tests mostly show cell viability above 70%, proving that the extracted collagen is non-toxic and meets the standard collagen criteria as a raw material for medical biomaterials.

Item Type:	Thesis (Masters)
Uncontrolled Keywords:	Kolagen, ekstraksi, tulang bovine, ultrasonik, asam asetat, biomaterial, collagen, extraction, bovine bone, ultrasonic, acetic acid, biomaterials
Subjects:	Q Science > QD Chemistry > QD251.2 Chemistry, Organic. Biochemistry Q Science > QD Chemistry > QD63 Extraction Q Science > QH Biology > QH541.15.T68 Toxicity testing R Medicine > R Medicine (General) > R857.M3 Biomedical materials. Biomedical materials--Testing.
Divisions:	Faculty of Industrial Technology and Systems Engineering (INDSYS) > Material & Metallurgical Engineering > 27101-(S2) Master Thesis
Depositing User:	Lailyta Shafira Amalia
Date Deposited:	27 Jan 2026 05:08
Last Modified:	27 Jan 2026 05:08
URI:	http://repository.its.ac.id/id/eprint/130433

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