Vì vậy, trong nghiên cứu này tôi đã tiến hành nghiên cứu sản xuất dầu cá hồi từ lườn cá bằng các phương pháp khác nhau và ứng dụng enzym lipase mủ đu đủ thủy phân dầu cá thu được để phục
TỔNG QUAN
Tổng quan về cá hồi
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Hình 1.1 Cá hồi Đại Tây Dương [6]
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Hình 1.2 Các loại cá hồi Thái Bình Dương [7]
• Cá hồi Chum (Oncorhynchus keta) có chiều dài khoảng 58cm, trọng lượng 15,9kg và tuổi thọ tối đa là 7 năm
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• Cá hồi Sockeye (Oncorhynchus nerka) có chiều dài khoảng 58cm, trọng lượng 7,7kg và tuổi thọ tối đa là 8 năm
• Cá hồi Steelhead (Oncorhynchus mykiss) có thân thon dài, 10 – 12 tia vây lưng,
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1.1.3 Thành phần dinh dưỡng của cá hồi
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Bảng 1.1 Thành phần dinh dưỡng có trong 100 g Cá Hồi Na Uy
(Nguồn: Viện Nghiên cứu Dinh dưỡng và Hải sản Na Uy)
Thành phần dinh dưỡng Đơn vị đo lường Hàm lượng dinh dưỡng trong mỗi 100g cá hồi Na Uy
1.1.4 Tình hình nuôi trồng, khai thác và chế biến cá hồi
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Tổng quan về dầu cá hồi
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*Một số tác dụng của dầu cá hồi:
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- Điều trị viêm khớp: dầu cá hồi, trong đó có chứa các acid béo chống viêm omega-
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Tổng quan về đu đủ
Cây đu đủ (danh pháp khoa học: Carica papaya) là một cây thuộc họ đu đủ papayaceae
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1.3.2 Thành phần hóa học và giá trị sử dụng của đu đủ
Bảng 1.2 Thành phần hóa học của thịt quả đu đủ chín [18]
Thành phần hóa học Hàm lượng % Thành phần hóa học Hàm lượng
Lipid 0,1 Vitamin PP 0,33 mg Đường 8,3 Muối khoáng:
+ Canxi + Sắt + Magie + Kali + Natri
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Mủ đu đủ nằm trong các ống dẫn mủ trải rộng khắp toàn bộ cây, ngoại trừ rễ Quả xanh vào khoảng 10 tuần tuổi chứa nhiều mủ nhất [23]
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2000) Ngoài ra nó còn được dùng trong sản xuất mỹ phẩm (Singh và Sirohi 1977; Knight 1980), công nghệ thực phẩm, công nghiệp thuốc tẩy…[23]
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Tổng quan về enzym lipase
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Hình 1.5 Mô hình enzym lipase [29] Hình 1.6 Cấu trúc của enzym lipase [30]
1.4.4 Cơ chế động học xúc tác của enzym lipase
Trong quá trình xúc tác, một đặc điểm quan trọng của lipase là có sự thay đổi về dạng không gian của vùng (trung tâm) hoạt động
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Hình 1.7 Mô hình cơ chế xúc tác của lipase ở bề mặt tiếp xúc giữa hai pha dầu – nước
E : Là lipase hòa tan có hoạt tính
E* : Là lipase hoạt động được hấp thu
S : Là cơ chất không tan trong nước
E*S: Là các phức hệ cơ chất lipase
P : Là sản phẩm phản ứng
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Lipase xúc tác cho một phản ứng hóa học qua ba bước (Martinelle, Hult, 1994)
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Hình 1.8 Sự hoạt hóa enzyme
- Bước 2: khi lipase gắn vào bề mặt phân pha thì vòng xoắn helix di chuyển, cho phép cơ chất gắn vào trung tâm hoạt động (Hình 1.9)
Hình 1.9 Hình thành phức hợp enzyme - cơ chất
- Bước 3: tạo thành sản phẩm, còn enzyme được giải phóng ra dưới dạng tự do
1.4.5 Một số phản ứng đặc trưng được xúc tác bởi enzym lipase
Enzym lipase xúc tác cho nhiều phản ứng hóa học khác nhau và thường được chia làm 2 loại: phản ứng thủy phân và phản ứng tổng hợp
- Phản ứng tổng hợp: được chia thành 4 nhóm nhỏ sau: a) Este hóa (Esterification)
RCOOH + R’OH RCOOR’ + H2O b) Este hóa tương hỗ (Interesterification)
RCOOR’ + R”COOR* RCOOR* + R”COOR’ c) Rượu phân (Alcoholysis)
RCOOR’ + R”OH RCOOR” + R’OH d) Axit phân (Acidolysis)
Trong đó 3 nhóm b, c và d thường được gọi chung là là transesterification [31].
Tổng quan về iso-octan
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Bảng 1.3 Một số tính chất vật lí đặc trưng của iso-octan [32]
Khối lượng phân tử 114.232 g.mol −1
Tỉ trọng 0,692 g cm -3 Điểm nóng chảy -107,38 °C; -161,28 °F; 165,77 K Điểm sôi 99,30 °C; 210,74 °F; 372,45 K Áp suất hơi 5,5 kPa (ở 21 °C)
UV-vis (λtối đa) 210nm
Iso-octan là đồng phân của octan nên có đầy đủ tính chất hóa học của một ankan Một số phản ứng hóa học đặc trưng của iso-octan:
- Iso-octan là một thành phần của xăng, thường được dùng làm chất chuẩn trong xác định tính chống kích nổ của xăng (trị số octan) [32]
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Tình hình nghiên cứu
1.6.1 Tình hình nghiên cứu trên thế giới
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1.6.2 Tình hình nghiên cứu trong nước
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Cơ sở lựa chọn phương pháp nghiên cứu
1.7.1 Một số phương pháp thu dầu cá
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Phương pháp dùng nhiệt: sử dụng nhiệt độ cao để phá vỡ màng tế bào của nguyên liệu, dầu từ các mô dự trữ sẽ chảy ra
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1.7.2 Thủy phân dầu cá hồi bằng enzym lipase mủ đu đủ
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ĐỐI TƯỢNG VÀ PHƯƠNG PHÁP NGHIÊN CỨU
Đối tượng, hóa chất và thiết bị nghiên cứu
Lườn cá hồi được mua ở Công ty TNHH chế biến thực phẩm D&N tại 01 Nguyễn Phục, Thọ Quang, Sơn Trà, Đà Nẵng
Hình 2.1 Lườn cá hồi thu mua tại Công ty TNHH chế biến thực phẩm D&N
2.2.1.2 Lipase thô từ mủ đu đủ a) Thu nhận mủ đu đủ
Mủ đu đủ: được thu từ một số vườn đu đủ trồng tại huyện Đại Lộc, tỉnh Quảng Nam
Hình 2.2 Vườn thu nhận mủ đu đủ
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+ Mủ đu đủ lấy ở quả xanh còn trên cây
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+ Không nên trộn mủ khô với mủ tươi vì nó làm giảm chất lượng
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+ Khi lấy mủ cần tránh không cho các chất bẩn hoặc côn trùng lẫn vào
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Hình 2.3 Sơ đồ thu chế phẩm enzym lipase thô
2.1.2 Hóa chất, dụng cụ và thiết bị sử dụng
Bảng 2.1 Dụng cụ, hóa chất và thiết bị sử dụng
Iso – octan, acid oxalic, KOH, đietyl ete,
Na2SO4, cồn 96%, thuốc thử phenolphtalein
1%, NaCl, NaH2PO4, Na2HPO4
- Tất cả các hóa chất sử dụng đều đạt độ tinh khiết phân tích, xuất xứ từ Việt Nam và Trung Quốc
- Dụng cụ thiết bị sử dụng tại phòng thí nghiệm trường Đại học Bách khoa Đà Nẵng và trường đại học Duy Tân
Các loại cốc thủy tinh: 100ml, 250ml, 500ml
Các loại ống đong: 50ml ,100ml, 250ml
Bình tam giác: 100ml, 250ml
Bình định mức 100ml, 500ml, 1000 ml
Cuvet, phễu chiết, buret, pipet Đũa khuấy thủy tinh, bóp cao su, ống sinh hàn
Cân kỹ thuật điện tử, cân phân tích 4 số, máy ly tâm EBA 20 Hettich (Đức), máy ép trục vít, máy khuấy từ
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Hình 2.4 Máy ép trục vít Hình 2.5 Cân phân tích 4 số
Hình 2.6 Máy ly tâm Hình 2.7 Cân kỹ thuật điện tử
Phương pháp nghiên cứu
2.2.1 Nghiên cứu sản xuất dầu cá hồi bằng phương pháp xay – nấu a) Sơ đồ tách dầu dự kiến
Hình 2.9 Sơ đồ quy trình sản xuất dầu bằng phương pháp xay – nấu b) Thuyết minh
• Nguyên liệu: lườn cá hồi sau khi mua về được đem đi rã đông và xử lí sơ bộ để loại bỏ phần vây cá
- Mục đích: phá vỡ cấu trúc tế bào, làm cho kích thước nguyên liệu nhỏ và đồng đều tạo điều kiện cho dầu dễ thoát ra
- Phương pháp thực hiện: lườn cá sau khi xử lí được cho vào máy xay sinh tố để xay nhỏ
- Mục đích: tiếp tục phá vỡ cấu trúc của tế bào, phá vỡ liên kết hóa học giữa lipid
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- Phương pháp thực hiện: lườn cá sau khi xay được bổ sung thêm nước (tỉ lệ 2:1 so với nguyên liệu) để tiến hành nấu trên bếp điện
- Mục đích: loại bỏ phần bã, protein đông tụ…để thu dịch lọc, tạo điều kiện thuận lợi cho các công đoạn sau
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- Mục đích: loại bỏ nước và các tạp chất có trong dịch lọc để thu phần dầu
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- Mục đích: loại bỏ triệt để nước và cặn có trong dầu
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• Xác định thời gian nấu
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- Mỗi thí nghiệm được bố trí với 3 lần lặp
• Khảo sát ảnh hưởng của muối NaCl đến hiệu suất thu hồi dầu cá
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- Mỗi thí nghiệm được bố trí với 3 lần lặp
2.2.2 Nghiên cứu sản xuất dầu cá hồi bằng phương pháp cắt khúc – ép a) Sơ đồ tách dầu dự kiến
Hình 2.10 Sơ đồ quy trình sản xuất dầu bằng phương pháp cắt khúc – ép
Ly tâm (3000 vòng/phút, 10 phút) Ép
Cặn Nước, cặn thô b) Thuyết minh
• Nguyên liệu: lườn cá hồi sau khi mua về được đem đi rã đông và xử lí sơ bộ để loại bỏ phần vây cá
- Mục đích: làm cho kích thước nguyên liệu nhỏ và đồng đều, phá vỡ 1 phần cấu trúc của tế bào, tạo điều kiện thuận lợi cho quá trình nấu
- Phương pháp thực hiện: lườn cá sau khi xử lí được đem đi cắt thành những phần có kích thước bằng nhau
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- Phương pháp thực hiện: lườn cá sau khi cắt khúc được bổ sung thêm nước (tỉ lệ 2:1 so với nguyên liệu) để tiến hành nấu trên bếp điện
- Mục đích: tách phần bã và phần dịch lọc
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- Mục đích: thu triệt để lượng dầu còn lại trong bã
- Phương pháp thực hiện: phần bã thu được sau lọc được đem đi ép bằng máy ép trục vít
- Mục đích: loại bỏ nước và các tạp chất có trong dịch lọc và dịch ép để thu phần dầu
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- Mục đích: loại bỏ triệt để nước và cặn có trong dầu
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- Mỗi thí nghiệm được bố trí với 3 lần lặp
- Lưu ý: thời gian nấu và lượng muối bổ sung vào quá trình lắng ở phương pháp này chính là điều kiện đã chọn ở phương pháp trên
2.2.3 Lựa chọn phương pháp sản xuất dầu cá và xác định hiệu suất thu hồi dầu cá thô Để lựa chọn phương pháp sản xuất dầu cá cho hiệu suất lớn nhất, tiến hành so sánh lượng dầu thu được ở các điều kiện thích hợp của 2 phương pháp, chọn ra phương pháp thu được nhiều dầu nhất Sau đó, tiến hành thí nghiệm theo phương pháp đã chọn ở các điều kiện phù hợp đó với mẫu có khối lượng lớn hơn (khoảng 500g), từ đó tính được hiệu suất thu hồi dầu
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Hiệu suất thu nhận = Khối lượng dầu cá
2.2.4 Phương pháp đánh giá chỉ tiêu chất lượng của dầu cá thô
- Xác định chỉ số axit theo TCVN 6127:1996; ISO 660:1983 [44]
- Xác định chỉ số xà phòng hóa theo TCVN 6126:2007; ISO 3657:2002 [45]
- Xác định chỉ số este: chỉ số este = chỉ số xà phòng – chỉ số axit
2.2.5 Phương pháp xác định thành phần axit béo có trong dầu cá hồi