臺灣博碩士論文加值系統

第一部份：
本研究之目的在探討吳郭魚片的血合肉在貯藏中自然色澤的保持，取吳郭魚血合肉及其肌紅蛋白溶液調整成不同pH值後，進行冰藏及凍藏之模擬試驗。另外，於吳郭魚片加工廠，以即殺死亡與掙扎死亡之吳郭魚為原料，製成魚片後進行凍藏實驗，探討pH值對血合肉顏色的影響。
首先，從吳郭魚片的血合肉萃取其肌紅蛋白溶液，以磷酸緩衝溶液（5.0～8.0）調整至不同pH值後，分別於0°C、－20°C貯藏，各樣品的氧化型肌紅蛋白（MetMb%）皆會隨著貯藏期間的增加而逐漸上升，其中，又以pH值低於6.3的溶液上升趨勢最快，於0°C冰藏時，2天內MetMb就已超過50%。凍藏於－20°C，在14天內就會超過50%，色澤CIE a*值皆會隨著冰藏或凍藏時間的延長而逐漸降低，以pH值低於6.3的肌紅蛋白溶液下降的速度較快，同時可觀察到肌紅蛋白溶液的明顯褐變，但pH值高於6.5之溶液在凍藏期間並無明顯之顏色變化（p＞0.05），顯示吳郭魚肌紅蛋白溶液發生自動氧化的臨界pH值應在6.3～6.5之間。
其次，吳郭魚片的血合肉經過10%、5%乳酸或10%碳酸鈉浸泡處理，再經真空包裝後，分別於0°C冰藏或－20°C凍藏。於冰藏或凍藏期間，血合肉中的MetMb%均會隨貯存時間之增加而逐漸上升，其中，又以10%乳酸處理組（pH 4.98）上升速度為最快，其次為5%乳酸處理組（pH 5.72）、控制組（pH 6.36），最慢者為10%碳酸鈉處理組（pH 6.72）；同時，CIE a*值的下降亦是以乳酸處理組為最明顯。外觀顏色方面，在冰藏環境下，10%乳酸處理組在第1天即可觀察到明顯的褐變，以5%乳酸處理者則在第1天可觀察到稍微褐變，而控制組在第9天才有明顯褐變，10%碳酸鈉處理組在第11天仍未發現褐變的現象。在凍藏環境下，也是以乳酸處理者褐變速度較快，碳酸鈉處理者褐變較慢。又，經過碳酸鈉處理之魚片若先凍結後再真空包裝，則此種「凍結後包裝」的魚片顏色優於傳統之「包裝後凍結」。
最後，將吳郭魚分為掙扎組、未掙扎組以及對照的CO處理組。掙扎組是在室溫下讓活魚於空氣中掙扎至死，未掙扎組係先將魚冰昏後立即宰殺製成魚片，CO處理組則是依工廠既定流程製成魚片且經CO處理，三組魚片之pH值依序分別為pH 6.5、7.0及6.6。以MetMb%來看，掙扎組之魚片於－20°C凍藏期間，MetMb%上升速度明顯快於未掙扎組（p＜0.05），凍藏至60天時，未掙扎組MetMb%只有22.1%，但掙扎組的魚片凍藏至45天時即已達到41.3%。血合肉的CIE a*值下降速度以及色差計反射光譜630 nm/580 nm比值的下降速度皆是以掙扎組為較快。CO處理的對照組之魚片在－20°C凍藏期間，其MetMb%並無明顯變化，魚片CIE a*值明顯高於掙扎組與未掙扎組（p＜0.05）。因此若依我國政府規定禁用CO氣體於魚片的保色處理時，則盡量減少魚肉在死亡後pH值的下降，另外，採用先凍結再真空包裝的方法，將有助於魚片血合肉在凍藏期間色澤的保持。
第二部份：
本研究將莫三比克種吳郭魚(Oreochromis mossambicus)、歐利亞種吳郭魚(O. aureus)、尼羅種吳郭魚(O. niloticus)及尼羅種與歐利亞種雜交吳郭魚(O. niloticus (♀) × O. aureus (♂))分別由其血合肉中抽出Total RNA，先逆轉錄成cDNA，再選殖出4種Myoglobin轉譯部分之基因序列。其次再由抽出之Total RNA以RLM-RACE (RNA Ligase Mediated Rapid Amplification of cDNA Ends)定出莫三比克吳郭魚Mb全部基因序列。
不同吳郭魚種間的肌紅蛋白胺基酸具有高度的保留區域，其序列之相似度高達100%；再將吳郭魚Mb胺基酸序列與NCBI database中已註冊之大目鮪（Bigeye tuna）、黑鮪（Bluefin tuna）、鯖魚（Chub mackerel）、黑皮旗魚（Atlantic blue marlin）之Mb的胺基酸序列進行比對，其相似度分別為81.6%、81.6%、74.8%、76.9%。
另外，比較吳郭魚與大目鮪Mb之α螺旋片段（A、B、C、E、F、G、H區域）及非螺旋區域之胺基酸序列，發現C螺旋區域、F螺旋區域、F-G非螺旋區域與G-H非螺旋區域，彼此相似度均為100%；而在B螺旋區域、H螺旋區域、與C-E非螺旋區域則相似度較低，分別只有73.3%、65.0%及50.0%。其餘相似度介於82-93%。以RACE方式定出莫三比克吳郭魚Mb基因全部序列發現有797 bp，其中可轉譯出147個胺基酸。
本研究再從不同吳郭魚種之血合肉萃取其肌紅蛋白溶液後，於25°C的水浴下測定其自動氧化速率，結果也發現4種吳郭魚的肌紅蛋白自動氧化速率常數極為相近，顯示不同種的吳郭魚Mb構造具有高度保留性，自動氧化速率也幾乎一樣。

Part 1:
This study was aimed to maintain the nature color of tilapia dark muscle during storage. Tilapia dark muscle and its myoglobin extract were subjected to estimate color changes (in a model test) during iced and frozen storage after adjusting pH to various values. In addition, alive tilapia sacrificed with or without struggle was filleted and frozen as the ordinary procedure in a processing factory to investigate the effect of pH on color of the tilapia fillets.
The myoglobin extracts were prepared with phosphate buffer at pH 5.0-8.0 and individually stored at 0°C or −20°C. The results showed that metMb formation ratio (MetMb%) of all samples increased gradually during iced or frozen storage. We found that formation of metMb in the samples with pH lower than 6.3 was accelerated and the MetMb% of the samples stored at 0°C and −20°C exceeded 50% within 2 days and 14 days, respectively. Moreover, their color CIE a* values decreased gradually during both iced and frozen storage and they changed faster as pH was lower than 6.3. Also significant browning in myoglobin solutions were observed when their pH values were lower than 6.3. However, there was no significant browning occurred when solution pH was higher than 6.5. It indicates that the critical pH for autoxidation of tilapia myoglobin (in solution) is probably at the range of 6.3-6.5.
Subsequently, the effect of acids on metMb formation in tilapia dark muscle was investigated. Tilapia dark muscle samples soaked in 10, 5% lactic acid, or 10% sodium carbonate were vacuum packaged and separately stored at 0°C or −20°C. MetMb% of all samples increased gradually during iced or frozen storage. It indicated that the autoxidation rate of the groups treated with 10% lactic acid (pH 4.98) and treated with 5% lactic acid (pH 5.72) were the fastest and second, respectively. Besides, that of control (pH 6.36) and 10% sodium carbonate group (pH 6.72) are the third and slowest, respectively. Meanwhile, the group treated with lactic acids showed the most significant in the decrease of CIE a* value. A significant browning was observed on the appearance of the group treated with 10% lactic acid after 12-h iced storage and the group treated with 5% lactic acid displayed a slightly browned color after 1-d iced storage. Browning on the appearance of the control was not observed until the 9th day. However, no noticeable browning appearance on the group treated with 10% sodium carbonate was seen on the 11th day. During froze storage, the group treated with lactic acid was very subjected to browning while the group treated with sodium carbonate enabled to retard the browning well. Moreover, the fillets treated with 10% sodium carbonate were carried out vacuum packing after freezing, frozen and then packed procedure, and show a better color display than those made by the traditional “packed and then frozen procedure”.
In addition to effect of acid solutions, the effects of the slaughter methods and CO treatment on metMb formation in tilapia dark muscle were also inspected. Tilapia fillets were obtained from various treatments. They were (1) struggle group (alive tilapia struggling till death), (2) instantly killed group (iced alive tilapia and sacrificed immediately), and (3) CO treated group (fillet prepared in a factory with CO gas treatment) and the pH of three groups was 6.5, 7.0 and 6.6, respectively. MetMb% in fillets of the struggle group increased faster, 41.3% on the 45th day, while that of the instantly killed group was 22.1% on the 60th day during frozen storage. Comparing to other groups, the dark muscle of fillets from the struggle group exhibited a faster decrease in both CIE a* value and 630 nm/580 nm ratio. The group treated with CO gas showed no significant change in MetMb% during storage at −20°C but it had higher CIE a* value than other two groups（p＜0.05）. However, the CO gas treatment for fish fillets is prohibited by Taiwanese law. To maintain nature color of dark muscle during frozen storage, the decline in pH of fish during post-harvest should be minimized. Moreover the “frozen and then packed procedure” would be beneficial to color preservation of tilapia fillets.
Part 2:
In order to compare the amino acid sequence of Mb between different tilapia species, total RNA were extracted from dark muscles of mossambicus tilapia (Oreochromis mossambicus), aureus tilapia (O. aureus), niloticus tilapia (O. niloticus) and tilapia hybrid (O. niloticus (♀) × O. aureus (♂)). Reversely transcribed to cDNA, and then gene sequence of myoglobin coding region segments were cloned. Moreover, total length of Mb gene sequence of mossambicus tilapia were determined from total RNA by RLM-RACE (RNA Ligase Mediated Rapid Amplification of cDNA Ends).
Deduced amino acid sequences of Mb were completely conserved in different tilapia species, and the identities was 100%. Comparing the alignment of the deduced amino acid sequences of myoglobin in the different tilapia with bigeye tuna, bluefin tuna, chub mackerel, and Atlantic blue marlin, we found that their identities were 78, 78, 75 and 77%, respectively.
Furthermore, assessment of the amino acid sequence of myoglobin in the α-helical segments (A, B, C, E, F, G, H) and intersegmental regions between tilapia and bigeye tuna, it indicated that the similarities among α-helical segment C and segment F, as well as intersegment region F-G and region G-H were 100%. Moreover, the sequence identities of α-helical segments B and segment H and intersegment region F-G were quite low only 73.3, 65.0 and 50.0, respectively. Identities of the others segment and intersegment were 82-93%. On the other hand, as nucleotide sequence of mossambicus tilapia myoglobin were determined by RACE, there were 797 base pairs, encoding 147 amino acids, were found in the gene sequence of tilapia Mb.
In addition, autoxidation of myoglobin extract solutions from dark muscle of different tilapia species were measured at 25°C. The results showed that autoxidation rate constants of Mb from different tilapia species were very similar. It suggests that autoxidation rate constants of tilapia Mb are similar due to highly conserved structures in different species.

目 錄
頁次
目錄……………………………………………………………………… I
表目錄…………………………………………………………………… V
圖目錄…………………………………………………………………… VI
相片目錄………………………………………………………………… IX
誌謝……………………………………………………………………… X
Part 1
第一部份：調整吳郭魚血合肉pH值對其貯藏期間顏色的影響……… 01
中文摘要………………………………………………………………… 02
英文摘要………………………………………………………………… 04
壹、前言………………………………………………………………… 06
貳、文獻整理…………………………………………………………… 08
ㄧ、吳郭魚……………………………………………………………… 08
（ㄧ）吳郭魚簡介……………………………………………………… 08
（二）吳郭魚的養殖…………………………………………………… 08
（三）冷凍吳郭魚片的加工…………………………………………… 10
（四）吳郭魚片的血合肉（Dark muscle）及品質指標……………… 10
二、肌紅蛋白…………………………………………………………… 11
三、影響肌肉變色的因素……………………………………………… 14
（一）pH值…………………………………………………………… 14
（二）溫度……………………………………………………………… 15
（三）屠宰前的狀況及屠宰後的處理………………………………… 15
（四）調氣包裝與還原劑之使用……………………………………… 16
（五）以ㄧ氧化碳（CO）氣體進行發色處理………………………… 17
参、材料與方法…………………………………………………………… 18
一、材料………………………………………………………………… 18
（一）實驗原料………………………………………………………… 18
1. 模擬試驗之吳郭魚片…………………………………………… 18
2. 工廠實驗之吳郭魚片…………………………………………… 18
（二）藥品……………………………………………………………… 18
1. 氧化型肌紅蛋白比例之測定…………………………………… 18
2. 肌紅蛋白濃度之定量…………………………………………… 19
3. 揮發性鹽基態氮之測定………………………………………… 19
4. 臭氧濃度之定量………………………………………………… 19
5. CO殘存量之檢測………………………………………………… 20
二、方法………………………………………………………………… 20
（一）實驗設計………………………………………………………… 20
（二）實驗方法………………………………………………………… 20
1. 肌紅蛋白（Mb）溶液之製備…………………………………… 20
2. 模擬試驗魚片之處理…………………………………………… 20
3. 臭氧水之製備及定量…………………………………………… 24
4. 分析方法………………………………………………………… 25
（1）血合肉中氧化型肌紅蛋白百分比（MetMb%）之測定… 25
（2）肌紅蛋白（Mb）溶液濃度之測定………………………… 25
（3）pH值之測定………………………………………………… 26
（4）揮發性鹽基態氮之測定…………………………………… 26
（5）CO殘存量之檢測………………………………………… 26
（6）吳郭魚片血合肉表面色澤及630 nm/580 nm比值之測定
………………………………………………………………
28
（7）統計分析…………………………………………………… 28
肆、結果與討論…………………………………………………………… 29
一、肌紅蛋白萃取液之模型試驗……………………………………… 29
（一）Mb萃取液在0°C冰藏期間顏色之變化……………………… 29
1. 氧化型肌紅蛋白百分比（MetMb%）…………………………… 29
2. 溶液色澤………………………………………………………… 29
3. 肌紅蛋白之濃度………………………………………………… 32
（二）Mb萃取液在－20°C凍藏期間顏色之變化…………………… 33
1. 氧化型肌紅蛋白百分比（MetMb%）…………………………… 33
2. Mb萃取液色澤…………………………………………………… 34
二、吳郭魚片之模擬試驗……………………………………………… 36
（一）魚片冰藏實驗…………………………………………………… 36
1. 氧化型肌紅蛋白百分比（MetMb%）…………………………… 36
2. 色澤方面………………………………………………………… 38
3. 氧化型肌紅蛋白百分比（MetMb%）與a*值、630 nm/580 nm
比值的相關性……………………………………………………
42
4. 鮮度變化………………………………………………………… 42
（二）魚片凍藏實驗…………………………………………………… 45
1. 氧化型肌紅蛋白百分比（MetMb%）…………………………… 45
2. 色澤方面………………………………………………………… 46
3. 氧化型肌紅蛋白百分比（MetMb%）與a*值、630 nm/580 nm
比值的相關性……………………………………………………
51
4. MetMb%達50%時的貯藏天數與pH值之間的相關性………… 51
三、吳郭魚片加工廠之現場試驗…………………………………… 54
1. 氧化型肌紅蛋白百分比（MetMb%）…………………………… 54
2. 色澤方面………………………………………………………… 55
3. 氧化型肌紅蛋白百分比（MetMb%）與a*值、630 nm/580 nm
比值的相關性……………………………………………………
59
4. 一氧化碳（CO）殘存量………………………………………… 61
伍、結論…………………………………………………………………… 62

Part 2
第二部分：不同吳郭魚種肌紅蛋白之基因的探討……………………… 63
中文摘要………………………………………………………………… 64
英文摘要………………………………………………………………… 65
壹、前言…………………………………………………………………… 66
貳、文獻整理……………………………………………………………… 67
參、材料與方法…………………………………………………………… 69
一、材料………………………………………………………………… 69
（一）實驗原料………………………………………………………… 69
（二）藥品及儀器……………………………………………………… 69
1. 藥品……………………………………………………………… 69
2. 儀器……………………………………………………………… 70
二、方法………………………………………………………………… 72
（一）實驗設計………………………………………………………… 72
（二）實驗方法………………………………………………………… 75
1. 肌肉中全部核醣核酸（Total RNA）之萃取…………………… 75
2. 互補去氧核醣核酸（cDNA）之製備…………………………… 75
3. 肌紅蛋白基因之選殖、純化、轉形與定序……………………… 75
4. RLM-RACE (RNA Ligase Mediated Rapid Amplifcation of
cDNA Ends) ……………………………………………………… 79
5. 自動氧化速率之測定…………………………………………… 82
肆、結果與討論…………………………………………………………… 84
一、不同吳郭魚種肌紅蛋白基因之選值及定序……………………… 84
二、莫三比克吳郭魚肌紅蛋白全長之選值及定序…………………… 86
三、不同吳郭魚種自動氧化速率之測定……………………………… 89
伍、結論…………………………………………………………………… 94
陸、參考文獻……………………………………………………………… 95



表目錄

Part 1 頁次
表2-1、吳郭魚的產量、價值及養殖面積………………………………… 09
表4-1、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間血合肉之pH值的變化…………………………………… 37
表4-2、吳郭魚片普通肉在冰藏期間VBN之變化…………………… 44
表4-3、CO處理組於－20°C凍藏期間，CO殘存量之變化…………… 61

Part 2
表3-1、本實驗所用之吳郭魚種………………………………………… 69
表3-2、本實驗所用之引子……………………………………………… 71
表4-1、吳郭魚肌紅蛋白的胺基酸序列與其他不同魚種之相似度比較…………………………………………………………………88
表4-2、吳郭魚肌紅蛋白之α–螺旋片段及非螺旋區域之胺基酸序
列與大目鮪之相似度比較………………………………………88
表4-3、不同吳郭魚種其肌紅蛋白之自動氧化速率常數………………93



圖目錄

Part 1 頁次
圖2-1、肌紅蛋白（myoglobin）及原血紅素（heme）的構造………………12
圖2-2、肌紅蛋白在肌肉中的變化……………………………………… 13
圖3-1、吳郭魚血合肉之肌紅蛋白粗抽出液之模擬實驗……………… 21
圖3-2、吳郭魚片模擬實驗之處理流程圖……………………………… 22
圖3-3、吳郭魚片之工廠實驗的處理流程圖…………………………… 23
圖3-4、CO測定用之GC裝置系統……………………………………… 28
圖4-1、吳郭魚血合肉Mb萃取液調整至不同pH值後，在冰藏期
間之MetMb%的變化……………………………………………30
圖4-2、吳郭魚血合肉Mb萃取液調整至不同pH值後，在冰藏期
間之CIE L*、a*、b*值的變化…………………………………31
圖4-3、Mb萃取液（pH 7.0）在冰藏期間之肌紅蛋白濃度的變化……33
圖4-4、吳郭魚血合肉Mb萃取液調整至不同pH值後，在－20°C
凍藏期間之MetMb%的變化……………………………………34
圖4-5、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間血合肉之MetMb%的變化…………………………………37
圖4-6、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間血合肉之CIE L*、a*、b*值的變化………………………41
圖4-7、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間血合肉之630 nm/580 nm之比值的變化…………………42
圖4-8、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間MetMb%與a*值變化之關係………………………………43
圖4-9、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於冰藏
期間MetMb%與630 nm/580 nm比值變化之關係……………43
圖4-10、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於
－20°C凍藏期間血合肉之MetMb%的變化…………………46
圖4-11、以乳酸及碳酸鈉調整吳郭魚片血合肉之pH值後，於
－20°C凍藏期間血合肉之CIE L*、a*、b*值的變化………50
圖4-12、以乳酸及碳酸鈉調整吳郭魚片血合肉pH值後，於－20°C
凍藏期間血合肉630 nm/580 nm的比值………………………51
圖4-13、以乳酸及碳酸鈉調整吳郭魚片血合肉pH值後，於－20°C
凍藏期間MetMb%與a*值變化之關係………………………52
圖4-14、以乳酸及碳酸鈉調整吳郭魚片血合肉pH值後，於－20°C
凍藏期間MetMb%與630 nm/580 nm比值變化之關係………53
圖4-15、以乳酸及碳酸鈉調整吳郭魚片血合肉pH值後，於0°C
冰藏及－20°C凍藏，觀察MetMb%達50%時的貯藏天數
與pH值之間的相關性…………………………………………53
圖4-16、吳郭魚經過掙扎組或未掙扎組前處理做成魚片，於－20°C
凍藏期間之MetMb%的變化…………………………………55
圖4-17、吳郭魚經過掙扎組或未掙扎組前處理做成魚片，於－20°C
凍藏期間之L*、a*、b*值的變化…………………………… 56
圖4-18、吳郭魚經過掙扎組或未掙扎組前處理做成魚片，於－20°C
凍藏期間630 nm/580 nm比值的變化…………………………59
圖4-19、吳郭魚經過掙扎組或未掙扎組前處理做成魚片，於－20°C
凍藏期間MetMb%與a*值變化的關係………………………60
圖4-20、吳郭魚經過掙扎組或未掙扎組前處理做成魚片，於－20°C
凍藏期間MetMb%與630 nm/580 nm比值變化的關係………60

Part 2
圖3-1、Mb基因選殖實驗流程圖……………………………………… 72
圖3-2、RLM-RACE流程圖……………………………………………… 73
圖3-3、由不同吳郭魚種血合肉萃取出的肌紅蛋白溶液在25°C條
件下自動氧化之測定……………………………………………74
圖3-4、RLM-RACE流程之設計…………………………………………79
圖4-1、由吳郭魚血合肉所製備之cDNA以Mb (F) primer及Mb (R)
primer選殖肌紅蛋白基因轉譯區域之2.0% Agarose電泳圖…85
圖4-2、以pGEM-T-Easy為載體，吳郭魚血合肉Mb轉譯區域基因接
合至大腸桿菌JM109後，質體DNA的檢定……………………86
圖4-3、不同吳郭魚種肌紅蛋白之胺基酸序列與其他鯖科魚類進行
序列排比…………………………………………………………87
圖4-4、由吳郭魚血合肉所製備之cDNA以（a）5’ RACE Outer (F)、
TR51 (R)與3’ RACE Outer (F)、TR31(F) primers；（b）5’
RACE Inner (F)、TR52 (R)與3’ RACE Inner (F)、TR32(F)
primers選殖肌紅蛋白5’及3’非轉譯區域之2.0% Agarose電
泳圖………………………………………………………………90
圖4-5、莫三比克吳郭魚肌紅蛋白cDNA之核苷酸及胺基酸序列……91
圖4-6、莫三比克吳郭魚與NCBI資料庫之莫三比克吳郭魚
（AY522582）之核苷酸進行序列排比…………………………92
圖4-7、四種不同吳郭魚種的肌紅蛋白溶液在25°C條件下之自動
氧化作用…………………………………………………………93



相片目錄

Part 1 頁次
相片2-1、莫三比克吳郭魚（O. mossambicus）……………………………09
相片4-1、以1.0 M磷酸緩衝溶液調整Mb萃取液至pH 7.0，於冰
藏期間外觀顏色之變化………………………………………32
相片4-2、吳郭魚血合肉Mb萃取液調整至不同pH值後，在－20°C 凍
藏期間溶液顏色的變化………………………………………35
相片4-3、以10%乳酸調整吳郭魚片血合肉之pH值後，於冰藏期
間表面色澤的變化……………………………………………38
相片4-4、以5%乳酸調整吳郭魚片血合肉之pH值後，於冰藏期
間表面色澤的變化……………………………………………39
相片4-5、未處理的吳郭魚片血合肉，於冰藏期間表面色澤的變化…39
相片4-6、以10% Na2CO3調整吳郭魚片血合肉之pH值後，於冰藏期
間表面色澤的變化……………………………………………40
相片4-7、以10%乳酸調整吳郭魚片血合肉之pH值後，於－20°C
凍藏期間表面色澤的變化……………………………………47
相片4-8、以5%乳酸調整吳郭魚片血合肉之pH值後，於－20°C
凍藏期間表面色澤的變化……………………………………47
相片4-9、未處理之吳郭魚片血合肉，於－20°C凍藏期間表面色
澤的變化………………………………………………………48
相片4-10、以10% Na2CO3調整吳郭魚片血合肉之pH值後，於
－20°C凍藏期間表面色澤的變化…………………………48
相片4-11、以10% Na2CO3調整吳郭魚片血合肉之pH值後，於
－20°C凍藏期間表面色澤的變化…………………………49
相片4-12、由正常加工所生產且經CO處理之吳郭魚片，於－20°C
凍藏期間表面色澤的變化……………………………………57
相片4-13、由經過掙扎處理之吳郭魚所生產之魚片，於－20°C凍
藏期間表面色澤的變化……………………………………58
相片4-14、由未經掙扎處理之吳郭魚所生產之魚片，於－20°C凍
藏期間表面色澤的變化……………………………………58

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