Accepted Manuscript

Change regularity of the characteristics of Maillard reaction products derived from

xylose and Chinese shrimp waste hydrolysates

Luyun Cai, Dongmei Li, Zhijian Dong, Ailing Cao, Hong Lin, Jianrong Li

PII: S0023-6438(15)30178-X
DOI: 10.1016/j.lwt.2015.09.007
Reference: YFSTL 4955

To appear in: LWT - Food Science and Technology

Received Date: 27 April 2015

Revised Date: 31 August 2015
Accepted Date: 2 September 2015

Please cite this article as: Cai, L., Li, D., Dong, Z., Cao, A., Lin, H., Li, J., Change regularity of
the characteristics of Maillard reaction products derived from xylose and Chinese shrimp waste
hydrolysates, LWT - Food Science and Technology (2015), doi: 10.1016/j.lwt.2015.09.007.

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1 Change regularity of the characteristics of Maillard reaction

2 products derived from xylose and Chinese shrimp waste hydrolysates

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4 Luyun Caia, Dongmei Lia, Zhijian Donga, Ailing Caob, Hong Linc, Jianrong Lia*

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5

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6 College of Food Science and Engineering, Bohai University, National & Local J

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7 oint Engineering Research Center of Storage, Processing and Safety Control Technolo

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8 gy for Fresh Agricultural and Aquatic Products, Food Safety Key Lab of Liaoning Pro

9 vince, Jinzhou, 121013, China

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b

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Xiaoshan Entry-Exit Inspection and Quarantine Bureau, 118 Tonghuizhong Road,
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11 Hangzhou, 311208, China
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12 College of Food Science and Engineering, Ocean University of China, Qingdao,

13 266100, China
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15 Corresponding Author: Jianrong Li

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16 * Tel./fax: +86 416 3400008. E-mail: [email protected]

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19 ABSTRACT
20 Chinese shrimp waste hydrolysates (CSWHs) with or without xylose were heated
21 with increasing time (0-180 min) at 115 °C, pH 8.0, to explored characteristics of
22 Maillard reaction and the rules for the formation of flavour in hydrolysates. The
23 results suggested that CSWHs could be modified by Maillard reaction and exhibited

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24 stronger pleasant flavour than thermal degradation at proper heating time. During
25 Maillard reaction, significant changes in Maillard reaction products (MRPs) were

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26 found, such as the accumulation of intermediate/browning/ fluorescent products, the
27 presentation of tempting reddish-brown, compared to Thermal degradation products

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28 (TDPs). Meanwhile, a series of volatile compounds and amino acids were found in
29 MRPs, and combined with sensory evaluation, the MRPs exhibited strong meaty and

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30 seafood aroma as well as umami and mouthfulness taste. Therefore, Chinese shrimp
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31 waste could potentially be used to produce flavoring essence with strong enticing
32 flavour and colour.
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33 Keywords: Chinese shrimp; Maillard reaction products; Flavour; Volatile compound;

34 Colour
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37 1. Introduction
38 Chinese shrimp (Fenneropenaeus chinensis), namely oriental shrimp, as an
39 important marine aquaculture species in China, is primarily distributed in the Bohai
40 Sea, Yellow Sea in China and the western ocean region of North Korea (Liu, 1959; Lu,
41 2009). Due to the delicacy, nutrient-rich, and a variety of vitamins as well as the

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42 essential trace elements included in protein, Chinese shrimp is one of the most
43 important aquatic processing products for high consumption. However, the resulting

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44 by-products contain abundant protein, mineral substance, phospholipids, and
45 astaxanthin, comprising approximately 35-45% of the whole shrimp weight.

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46 Moreover, improper disposal of these by-products may cause the serious environment
47 pollution with offensive odour and the wasting of resources. At present, the utilization

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48 of by-products mainly focused on recovery of protein fractions (Cao et al, 2014),
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49 utilization as animal feed (Cavalheiro et al, 2007), as well as further extraction of
50 chitosan and chitinous substance from by-products of shrimp (Ferrer et al, 1996). In
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51 addition, several studies have been made for shrimp flavors from shrimp wastes by
52 Maillard reaction (Zhang et al, 2006; Liu et al, 2009; Dong et al, 2013). However,
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53 their works just aimed at the optimization of the process of shrimp flavoring essence,
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54 few researches have been made to explore the regularity for the formation of aroma
55 and taste derived from Chinese shrimp waste hydrolysates during Maillard reaction.
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56 Maillard reaction plays an important role in food industry and occurs in the
57 thermal processing or storage of foods, which is also developed into an effective
method for protein modification (Su et al, 2011; Daniel et al, 2014). The Maillard
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59 reaction, also known as non-enzymic browning reaction, is an interaction between

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60 reducing sugars and amino groups on proteins, which produces a variety of advanced
61 compounds, intermediate products, browning products, and fluorescent products,
62 accompanied by the formation of markedly aroma and taste-enhancing properties.
63 These unique aroma are closely related to raw material in system, such as roasted
64 flavour in meat (Domínguez et al, 2014), and seafood flavour in marine products
65 (Kubota et al, 2002). In addition, Ogasawara et al (2006) suggested that the so-called
66 “Maillard peptide” isolated from the 1,000 to 5,000 Da peptide fractions on the basis
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67 of Maillard reaction in the hydrolysis system of soybean protein have strong

68 taste-enhancing (mouthfulness and continuity in umami solution and consomme soup)
69 properties. In this study, the Maillard reaction products (MRPs) were derived from
70 hydrolysate-xylose model system, and the changes in pH, colour, and sensory
71 evaluation of MRPs were examined to reveal the sensory characteristics, while the

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72 further insight of volatile compounds, intermediate products, browning products, and
73 fluorescent products, as well as the changes of free amino acids content and

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74 SDS-PAGE analysis, were to indicate the mechanism of Maillard reaction in
75 modification of proteins in CSWHs. The approach may have practical implications for

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76 the effective utilization of Chinese shrimp waste, and simultaneously, provide
77 theoretical basis for the production of flavoring essence.

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78 2. Material and methods
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79 2.1. Materials and chemicals
80 Chinese shrimp wastes were obtained from a local aquatic market in Jinzhou, China.
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81 After washing, the shrimp wastes were freeze-dried, crushed and the powder samples
82 were stored at -20 °C before use. Protamex (1.5 AU/g) was purchased from
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83 Novozymes Co., Ltd (Beijing, China) and stored at 4 °C until it was used for the
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84 hydrolysis experiments. The other chemicals used were of analytical grade or better
85 and obtained from Shanghai Chemical Reagent Co., Ltd. (Shanghai, China).
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86 2.2. Preparation of Chinese shrimp wastes hydrolysates (CSWHs)

87 The Chinese shrimp waste hydrolysates (CSWHs) were prepared according to
the method reported by Huong et al. (2012) with corresponding modifications. The
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89 lyophilized shrimp wastes powder was mixed with deionized water at a ratio of 1:25
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90 (w/v). The mixture was adjusted to pH 6.0 using 0.5 M NaoH or HCl, and
91 pre-incubated at 55 °C for 10 min. The enzymatic hydrolysis was initiated by adding
92 protamex at a percentage of 1.5% (w/w) with continuously stirring at 55 °C for 4 h.
93 After hydrolysis, the hydrolysates were heated at 85 °C for 10 min to inactivate the
94 enzymes, and the CSWHs were rapidly cooled to room temperature. The mixtures
95 were centrifuged at 5000 r/min for 10 min at 4 °C. Then the supernatants were
96 collected, lyophilized and stored at -20 °C until further use.
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97 2.3. Preparation of Maillard reaction products (MRPs) and thermal degradation

98 products (TDPs)
99 Maillard reaction products (MRPs) were prepared according to the method
100 described by Sun et al. (2010) with some modifications. The formulation was as
101 follows: Hydrolysates (solid content: 1.5%, 75 g), xylose (50 g). They were mixed

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102 absolutely and heated for 0, 10, 20, 30, 60, 90, 120, 150 and 180 min at 115 °C. After
103 reaction, the products were diluted to the original volume, cooled immediately by ice

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104 water and kept at 4 °C to obtain MRPs for further use. Thermal degradation products
105 (TDPs) were prepared from all the samples in the same way as described above

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106 without the addition of xylose.
107 2.4. Determination of pH, intermediate products and browning intensity

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108 The pH of MRPs and TDPs were measured using a PB-10 pH-meter (Sartorius,
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109 Gottingen, Germany) calibrated with buffer solutions of pH 4.0 and 10.0, respectively.
110 The intermediate products and browning intensity of MRPs and TDPs were
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111 determined by the method of Maillard et al. (2007). The samples were diluted by
112 50-fold with distilled water. The absorbance of samples was measured using a UV-vis
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113 spectrophotometer (Shimadzu UV-2550, Shimadzu Co., Kyoto, Japan) at 294 nm and
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114 420 nm, as an indication of the formation of intermediate products of nonenzymatic

115 browning and the brown polymers formed in more advanced stages for MRPs or
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116 TDPs, respectively.

117 2.5. Wavelength spectra of MRPs and TDPs
The MRPs and TDPs were diluted by 50-fold with distilled water. The
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119 wavelength spectra of MRPs and TDPs were recorded by a UV-vis spectrophotometer
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120 (Shimadzu UV-2550, Shimadzu Co., Kyoto, Japan), with the wavelength ranging
121 from 200 to 700 nm.
122 2.6. Determination of colour
123 The colour of MRPs and TDPs were determined using a Minolta Chroma Meter
124 CR 400 (Minolta, Osaka, Japan). The Chroma Meter was calibrated on the hunterLab
125 colour space system using a white tile (Y=86.8, x=0.3150, y=0.3212). Seven
126 milliliters of samples were pipetted into a 25 mm diameter test tube. The system
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127 provides the values of three colour components: L* (black-white component,

128 luminosity), the chromaticity coordinates, a* (red-green component) and b*
129 (yellow-blue component).
130 2.7. Fluorescence spectroscopy
131 The measurement of fluorescence spectra of MRPs and TDPs samples were

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132 operated by using a 970 CRT fluorescence spectrometer (Shanghai, China), according
133 to the method proposed by Jiang et al. (2014). The samples were diluted by 50-fold

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134 with Milli-Q water and excited at 386 nm. The spectrum of emission was recorded
135 from 350 to 550 nm with a slit of 2.5 nm for both the emission and the excitation.

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136 2.8. Sensory evaluation
137 Sensory evaluation panelists were trained to recognize basic tastes (umami,

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138 continuity, bitterness, sourness sweetness, mouthfulness and saltiness) and basic
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139 aroma (meaty, seafood, fishy, toasty and burnt). Sensory evaluation was carried out
140 according to the method reported by Ogasawara et al. (2006) with minor
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141 modifications. All the test solutions were adjusted to pH 7.0. Umami solution
142 consisted of 1.5% monosodium glutamate (MSG) and 0.5% sodium chloride (NaCl).
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143 β-Cyclodextrin was added to the umami solution as the control for sensory evaluation.
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144 The MRPs and TDPs (0.35%) were respectively dissolved in umami solution,
145 followed by heated to 60 °C in a water bath. Sixty milliliters of MRPs, TDPs and
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146 control solution were served in opaque disposable plastic cups at the same time,
147 respectively. Sensory evaluation was scored on a scale of 1-7, where 3 points was the
score given to the control solution. Each panel consisted of 16-20 panelists, half males
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149 and half females, 25-30 years old. All panelists had received training in descriptive
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150 sensory analysis (>20 h) and possessed experience in sensory profiles of various food
151 samples. Sensory evaluation sessions were conducted in an air-conditioned room (20
152 ± 2 °C) with panelists in separate booths.
153 2.9. Determination of the amino acids composition
154 The amino acids contents were determined using a full-automatic amino acid
155 analyzer (L-8900A, Hitachi, Tokyo, Japan). An appropriate pretreatment for the
156 samples was necessary before amino acids analysis, according to the method proposed
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157 by the researchers Song et al. (2013) with some modifications. 5 mL of samples were
158 added to a 25-mL volumetric flask, and 5% trichloroacetic acid (TCA) was added to
159 reach the scale to precipitate peptides or proteins, followed by incubation for 2 h at
160 4 °C. The solution was filtered through 0.22 µm filter membrane and the filtrate was
161 centrifuged at 14,000 r/min for 10 min. The supernatant was stored at 4 °C before

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162 injection. A calibration curve was obtained with standard amino acid mixture (Sigma,
163 St. Louis, MO, USA) and qualitative analysis was made on the basis of retention time

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164 and peak area of standard compounds.
165 2.10. Electrophoresis analysis

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166 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was
167 performed according to the method described by Martinez-Alvarenga (2014) with

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168 some modifications. Using a 4.0% stacking gel and a 15.0% separating gel, samples
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169 (50 µg) were separated using a PowerPac (Bio-Rad, China) with a constant voltage 80
170 V for 20 min and a constant voltage 120 V for about 2 h. After electrophoresis, the
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171 gels were stained using Coomassie brilliant blue G-250 (0.10%, w/v) in methanol
172 (15%, v/v) and acetic acid (5%, v/v) for about 2 h, and destained with methanol (30%,
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173 v/v) and acetic acid (10%, v/v) until a clear background was achieved.
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174 2.11. Analysis of volatile compounds

175 The determination of volatile compounds was carried out on an Agilent 7890 GC
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176 coupled to an Agilent 5975C quadropole mass selective detector (Agilent

177 Technologies, Thermo Trace DSQ II GC/MS, Palo Alto, CA, USA) and separated in
an HP-5MS capillary column (30 m×0.25 mm×0.25 µm internal diameter, 1 mm in
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179 film thickness). The mass spectrometer was carried out with electron impact mode at
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180 70 eV and the ion source temperature was set at 230 °C. The detector voltage was 350
181 V, and the scan range of 30-500 amu. 5 mL of samples were weighed into a 20-mL
182 headspace vial and sealed with a Teflon silicone septum. The vial was left at 55 °C in
183 a thermal block for 20 min to equilibrate the headspace. Then, an SPME fiber was
184 exposed to the headspace while maintaining the sample for 30 min. After extraction,
185 the compounds absorbed by the fiber was inserted into the GC injection port set at
186 230 °C for 3 min with a splitless injection mode. Helium was used as carrier gas with
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187 a flow rate of 1 mL/min. The GC oven temperature program for SPME procedure:
188 40 °C for 4 min, 40 °C-230 °C at 5 °C /min, and final temperature holding for 5 min.
189 Identification of flavour compounds by comparing mass spectrum data (RT/MS) of
190 the samples with the National Institute of Standards and Technology literature
191 database. Volatile compounds were quantified by calculating the peak area.

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192 2.12. Statistical analysis
193 All experimental data reported in this work were triplicate. Statistical analysis

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194 was performed using SPSS 13.0 (IBM, Armonk, USA) to determine significant
195 differences between samples. Differences at p < 0.05 were considered significant.

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196 3. Results and discussion
197 3.1. Changes in pH

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198 The changes of pH values derived from Maillard reaction (MR) and thermal
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199 degradation (TD) model systems as a function of the heating time are shown in Table
200 1. In the initial phase of heating, both of two model systems possessed the same
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201 values. However, the pH values in MR model system decreased rapidly as the heating
202 time increased, compared with a minor change of pH in TD model system, which was
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203 similar with the result reported by Jiang et al. (2012). Maillard reaction initiated
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204 between carbonyl groups of reducing sugars and free amino groups such as proteins,
205 peptides or amino acids, the consumption of the amino groups by MR could shift
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206 systems into more acidic condition (Eric et al, 2014). However, without reducing
207 sugars, the peptides degraded into smaller peptides or amino acids rapidly during TD
(Lan et al., 2010), which could contributed to the minor change of pH values in TD
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209 system.
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210 3.2. Changes in colour

211 Many studies have been reported that the colour of food was the reflection of
212 coloured natural products in the raw material and/or the generation of coloured
213 compounds from processing of food (Cristina et al, 2010). Table 1 depicted the
214 evolution of colour as the heating time increased for both MR and TD systems. The
215 luminosity (L*) decreased from 49.97 to 19.58 in MR system. The rapid decrease in
216 the L* value was due to the formation of brown pigments from MR. For the MR
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217 model system, the a* value ranged from -2.65 to -0.8, and the b* value increased from
218 13.73 to 26.85 within 60 min of heating, which suggested that MR model system
219 reflected more green and yellow characteristics. But after 60 min of heating, it
220 reflected more red and yellow characteristics. What was noteworthy was that the b*
221 and a* value reached the maximum value at 90 and 120 min, respectively, and then

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222 decreased as a function of heating time. The formation of yellow/brown coloration
223 could result from the action of the various intermediates or final products of the MR

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224 (Liu et al, 2004). The TD showed minor changes in L*, b* and a* value, which was
225 observed to reduced slightly as a function of heating time.

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226 3.3. Sensory properties
227 The TDPs showed slightly better of fishy, seafood, meaty flavour in Fig. 1a, and

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228 displayed continuity and mouthfulness in Fig. 1c compared to control sample, but
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229 they had no obvious change with increasing heating time. On the contrary, the MRPs
230 showed an increasing intensity of meaty and seafood flavour in Fig. 1b, and exhibited
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231 an increasing intensity of umami, continuity and mouthfulness in Fig. 1d with heating
232 time from 0 to 60 min. The reason might be related to that in the presence of xylose,
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233 the samples generated a great number of MRPs such as thiazoles, sulfides, aldehydes,
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234 and ketones, which made great contribution to the meaty flavour of hydrolysates
235 (Cross et al, 1965), and Maillard peptides, as good flavour enhancers, gave a
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236 characteristic umami, mouthfulness and continuity of the flavour (Ogasawara et al,
237 2006). Simultaneously, the fishy odour in samples weakened gradually. After 60 min
of heating, the MRPs displayed slight toasty, then this toasty might be changed into
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239 disagreeable burnt odour, that might be related to the increasing content of nitrogen
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240 containing compounds (Table 2). In the late stage of MR, the generation of pyrazines,
241 pyrroles, and miazines contributed to pleasant toasty flavour, even disagreeable burnt
242 odour, which could due to the caramelization of xylose or the overreaction of
243 materials. Undesirably, the meaty and seafood flavour in MRPs were terrible, mainly
244 due to the effect of undesirable flavours, for instance, burnt odour.
245 3.4. Comparison of volatile compounds of MRPs

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246 It was widely reported that the formation of volatile compounds was mostly due
247 to the difference of the composition of free amino acids, distribution of molecular
248 weight in protein hydrolysates, peptides composition and chemical configurations of
249 the above (Zhang et al, 1992). The volatile compounds from MR systems heated by
250 different time were shown in Table 2.

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251 Sulphur-containing volatile compounds are usually described as the meaty
252 flavour compounds in vegetables, cooked meat and other food. Dimethyl disulfide,

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253 dimethyl trisulfide and 2-acetylthiazole, were found in all samples, and their contents
254 increased initially and reduced afterwards. Dimethyl disulfide and dimethyl trisulfide

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255 possessed low odour thresholds and were reported to make a large contribution to the
256 overall sensory flavour in food, therefore, they exhibited both desirable and

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257 undesirable aromas, depending on their concentrations. 0.38-0.54% 2-acetylthiazole
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258 was detected in samples, and it was often detected in roasted, grilled and fried foods.
259 Other researchers found that sulphur-containing volatile compounds contributed to the
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260 roasty and meat-like flavour of foods (Domínguez et al, 2014).

261 Among the 13 nitrogen-containing compounds, 5 pyrazines, 1 pyrroles, 2
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262 diazines and 4 others compounds were identified in MR systems heated by 90, 150
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263 and 180 min, none was detected by 30 min. Nitrogen-containing compounds
264 generated in the final phase of MR, were responsible to the development of both
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265 flavour and colour compounds (Belitz et al, 2004). Pyrazines have been suggested as
266 being responsible for representative roasted, toasted, nutty and burnt of cooked foods,
and possessed extremely low odor threshold (Shimoda et al, 1990), resulted in an
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268 increasing pungent flavour as a function of heating time (Fig. 1b). Furans and its
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269 derivatives in samples provided roasted, toasted, cooked meat, coffee, and the specific
270 aroma of seafood (Yahya et al, 2014).
271 The result showed that furan non-S-containing compounds percentage increased
272 with increasing heating time. Similar result was found in a previous study that furans
273 content also increased with increasing temperature for sunflower hydrolysate during
274 Maillard reaction (Eric et al, 2014). Although a sharp increase content (2.02-39.41%)
275 of furans were detected in MR systems with increasing heating time, furans could not
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276 have prominent contribution to the flavour, which was attributed to their high odour
277 threshold values.
278 For aldehyde compounds, 3-methyl-butanal, hexanal, heptanal, benzaldehyde,
279 nonanal, benzeneacetaldehyde and decanal were found in all samples (Table 2).
280 Excepting for benzeneacetaldehyde and 3-methyl-butanal, the contents of other

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281 compounds showed a downward trend. Aldehydes often possessed low perception
282 thresholds, which made samples exhibit special aroma even at trace amounts. High

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283 percentage (18.63-32.80%) of 3-methyl-butanal was detected in all samples,
284 contributed to chocolate aroma and fragrance. The favorable aroma of aldehyde

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285 compounds such as methional (meat-like odour), benzaldehyde (cherry-like odour),
286 and benzeneacetaldehyde (rosy-like odour), can be supposed to provide samples

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287 special overall aroma.
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288 In addition, a number of ketones, alcohols, esters, acids, hydrocarbons, amines,
289 and phenols were found in all samples, their high perception thresholds made them
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290 contribute less to samples, but they reconciled the overall flavour of samples.
291 3.5. The development of fluorescence and UV-Vis spectra
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292 The development of fluorescence products during MR has been generally

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293 reported in sugar-amino group systems, and the formation of fluorescent compounds
294 was determined as early indicators of MR (Baisier et al, 1992). In addition, UV-Vis
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295 spectra have usually been used for characterization of the MRPs. The absorbance at
296 294 nm used to be an indication of the formation of an uncoloured compounds that
could be the precursor of MR (Kim et al, 2009), and the intensity of brown colour
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298 development (A 420 nm) was determined as an indicator of the final stage of MR
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299 (Serpen et al, 2009).

300 Fluorescence intensity of all samples were measured at an excitation wavelength
301 of 386 nm, emission wavelength was 450 nm, shown in Fig. 2a and 2b. Similar results
302 were found in MR of whey protein isolate conjugated with glucose reported by Liu et
303 al (2014). The fluorescence intensity of both TD and MR model system reduced at
304 386 nm, whereas only MR model system displayed marked increasing flurescence
305 intensity at approximately 465 nm with increasing heating time, the reason might be a
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306 long time heating force fluorescence compounds to accumulate gradually and reach a
307 maximum before being converted into visible brown pigments, slightly increased of
308 fluorescence intensity with heating time were found in TD model system.
309 UV-Vis spectra of MRPs measured over the range of 200-700 nm for TD and
310 MR model system as a function of heating time were presented in Fig. 2c and 2d,

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311 where absorption peak possessed similar shape. Moreover, each peak exhibited
312 absorption peak at approximately 265 nm. However, the maximum absorbance of MR

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313 model system had a more swiftly increase after 30 min of heating, compared with
314 slightly change within first 30 min of heating in Fig. 2c. Similarly, some researchers

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315 found that the UV-Vis spectra of MRPs from xylose had sharp absorption peak at
316 approximately 265 nm and exhibited dramatic higher absorption at 265 nm, and

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317 modification of amino acid by MR could enhance its absorption (Yu et al, 2012).
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318 Accordingly, the absorption maximum of MRPs shifted to 275 nm within 60-180 min
319 of heating. The redshift suggested that MR had possibly reached the final stage, while
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320 nearly no changes were found in hydrolysates heating alone.

321 3.6 Changes in intermediate products and browning intensity
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322 The values changes in absorbance at 294 nm and 420 nm derived from MRPs
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323 and TDPs as a function of heating time are shown in Table 1. Within the initial 20 min
324 of heating, the values changes in absorbance at 294 nm or 420 nm, were negligible, in
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325 both MR and TD model system. After 30 min of heating, the MR model system
326 obtained a marked increase in absorbance at 294 nm, which suggested the generation
of intermediate products, and the production of intermediate products increased as
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328 heating time progressed. However, from 30-60 min of heating, the accumulation of
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329 browning products derived from the MR model system, were gentle. Afterwards, the
330 values in absorbance at 420 nm increased rapidly, which revealed the deepening
331 extent of MR, a reasonable explanation could be that some intermediate products
332 polymerised and formed brown pigments in the late stage of MR. Similar increases in
333 absorbance at 294 and 420 nm were reported by Jung et al. (2014). By contrast, in the
334 absence of xylose, there were no significant changes with the values in absorbance at
335 both 294 and 420 nm.
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336 3.7. Changes in free amino acids

337 Free amino acids play an important role on the taste and the formation of flavour
338 compounds in samples. As indicated in Table 3, the total content of free amino acids
339 in TDPs decreased gradually within 60 min of heating, afterwards, its content tend to
340 flattened, that could be concluded to the TD of proteins or peptides as well as

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341 decarboxylation of amino acids. In addition, as the heating time prolonged, the TD
342 became more severe, which balanced the consumption and generation of amino acids

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343 in TDPs. While in MRPs, the total content of free amino acids had a sharpen decrease
344 during first 30 min of heating, and then it changed to be mild as well. It is worth

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345 mentioning that, aspartic acid (Asp) and glutamic acid (Glu) as important amino acids
346 for their umami taste, their content increasing in the later stage of heating, mainly due

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347 to the degradation of Maillard peptides, which contributed to strong umami taste as
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348 shown in Fig. 1d. The consumption of amino acids in MR made a great contribution
349 to aroma of samples, especially sulphur-containing amino acids such as Methionine
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350 (Met), and Cysteine (Cys) contributed to strong meaty flavour in samples, and the
351 results were related to the sensory evaluation in Fig. 1b and the Volatile compounds in
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352 Table 2.
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353 3.8 SDS-PAGE analysis

354 Changes in the molecular weight distribution of protein were evaluated by
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355 SDS-PAGE, in TD and MR model systems with increasing heating time, shown in Fig.
356 3a and 3b, respectively. In light of the high reactivity of low molecular weight
proteins during MR, we chose low range (4.1-66 kDa) protein marker (M) to prove
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358 the occurrence of MR between hydrolysates and xylose. The bands of hydrolysates
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359 alone (heating by 0 min) were numerous (Fig. 3a). As a function of heating time, a
360 great number of bands disappeared in TD systems, and the longer the heating time,
361 the more bands disappeared, only bands with 45-66 kDa and 16-17 kDa remained, the
362 reason might be related with the TD of protein. However, as heating by 180 min, all
363 bands were disappeared, it was reported that high molecular weight proteins degraded
364 into molecular weight proteins, peptides or amino acids during thermal treatment (Lan
365 et al, 2010). Similarly, a large number of bands disappeared as well in MR system
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366 with increasing heating time (Fig. 3b), which might be caused by both TD and MR of
367 protein. Particularly, bands with 45-66 kDa were completely disappeared in all MR
368 systems, but bands with 16-17 kDa remained only in systems by heating 30 and 60
369 min, which illustrated that TD was occurred more easily in high molecular weight
370 proteins than low molecular weight proteins, similar results were found by Lan et al.

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371 (2010).
372 4. Conclusions

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373 In the present study, Chinese shrimp waste hydrolysates (CSWHs) could be
374 modified by MR and exhibited stronger pleasant flavour or disagreeable flavour than

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375 TD, depending on the heating time. The Maillard reaction products (MRPs) by 60 min
376 showed stronger meaty and seafood aroma, as well as umami and mouthfulness taste

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377 compared to thermal degradation. Sulphur-containing compounds,
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378 nitrogen-containing compounds and aldehydes exhibited important contribution to
379 meaty and seafood aroma of MRPs. Moreover, Asp and Glu, whose contents
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380 increased in the later stage of heating, were considered essential contributors to
381 umami taste of MRPs. The results suggest that effective and reasonable control for the
D

382 process of MR derived from CSWHs would make great contribution to the production
TE

383 of flavoring essence, which achieved full utilization of Chinese shrimp waste.
384 Acknowledgments
EP

385 This study was supported by National Natural Science Foundation of China (314

386 01478; 31471639), National Postdoctoral Science Foundation of China (2015M57076

C

387 0), China Scholarship Council (201508210023), National Key Technology R&D Prog
AC

388 ram of China during the 12th Five-Year Plan Period (2012BAD29B06) and Liaoning

389 Provincial Science & Technology Project (201500098).

390 References
391 Baisier, W. M., & Labuza, T. P. (1992). Maillard browning kinetics in a liquid model system.

392 Journal of Agricultural and Food Chemistry, 40(5), 707-713.

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 ACCEPTED MANUSCRIPT

393 Belitz, H. D., Grosch, W., & Schieberle, P. (2004). Carbohydrates. Food Chemistry, 3rd ed.

394 Springer-Verlag, Berlin, Germany. pp 245-341.

395 Cao, W. H., Tan, C. Y., Zhan, X. J., Li, H. Y., & Zhang, C. H. (2014). Ultraviolet irradiation and

396 gradient temperature assisted autolysis for protein recovery from shrimp head waste.

397 Food Chemistry, 164, 136-141.

PT
398 Carvalho, D. O., Correia, E., Lopes, L., & Guido, L. F. (2014). Further insights into the role of

399 melanoidins on the antioxidant potential of barley malt. Food Chemistry, 160, 127-133.

RI
400 Cavalheiro, J. M. O., Souza, S. O., & Bora, P. S. (2007). Utilization of shrimp industry waste in

401 the formulation of tilapia (Oreochromisniloticus Linnaeus) feed. Bioresource

SC
402 Technology, 98(3), 602-606.

403 Cross, C. K., & Ziegler, P. (1965). A Comparison of the volatile fractions from cured and uncured

U
404 meat. Journal of Food Science, 30(4), 610-614.
AN
405 Domínguez, R., Gómez, M., Fonseca, S., Lorenzo, J. M. (2014). Effect of different cooking

406 methods on lipid oxidation and formation of volatile compounds in foal meat. Meat
M

407 Science, 97(2), 223-230.

408 Dong, Z. J., Li, D. M., Li, S. W., Li, X. P., Li, J. R., & Huang, H. (2013). Preparation of barbecue
D

409 shrimp flavoring essence by Maillard reaction from low-valued sea shrimp and its
TE

410 analysis by GC-MS and electronic nose. Science and Technology of Food Industry,

411 34(10), 101-107.

EP

412 Eric, K., Raymond, L. V., Abbas, S., Song, S. Q., Zhang, Y. T., & Kingsley, M. (2014).

413 Temperature and cysteine addition effect on formation of sunflower hydrolysate

C

414 Maillard reaction products and corresponding influence on sensory characteristics

415 assessed by partial least square regression. Food Research International, 57, 242-258.
AC

416 Ferrer, J., Marmol Z., Ramones, E., Garcia, H., Forster, C. F., & Paez, G. (1996). Acid hydrolysis

417 of shrimp-shell wastes and the production of single cell protein from the hydrolysate.

418 Bioresource Technology, 57(1), 55-60.

419 Huong, N. T. M., Raúl P. G., & Jean, P., B. (2012). Effect of diets containing tuna head

420 hydrolysates on the survival and growth of shrimp Penaeus vannamei. Aquaculture, 127,

421 324-325.

422 Jiang, Z. M., Wang, L. Z., Che, H. X., & Tian, B. (2014). Effects of temperature and pH on

15
 ACCEPTED MANUSCRIPT

423 angiotensin-I-converting enzyme inhibitory activity and physicochemical properties of

424 bovine casein peptide in aqueous Maillard reaction system. LWT-Food Science and

425 Technology, 59, 35-42.

426 Jin, W. G.., Wu, H. T., Li, X. S., Zhu, B. W., Dong, X. P., & Li, Y. (2014). Microstructure and

427 inter-molecular forces involved in gelation-like protein hydrolysate from

PT
428 neutrase-treated male gonad of scallop (Patinopecten yessoensis). Food Hydrocolloids,

429 40, 245-253.

RI
430 Jung, W. K., Park, P. J., Ahn, C. B., & Je, J. Y. (2014). Preparation and antioxidant potential of

431 maillard reaction products from (MRPs) chitooligomer. Food Chemistry, 145, 173-178.

SC
432 Kim, J. S., & Lee, Y. S. (2009). Study of Maillard reaction products derived from aqueous model

433 systems with different peptide chain lengths. Food Chemistry, 116(4), 846-853.

U
434 Kubota, S., Itoh, K., Niizeki, N., Song, X. A., Okimoto, K., & Ando, M. (2002). Organic taste
AN
435 active components in the hot-water extract of yellow tail muscle. Food Science and

436 Technology Research, 8(1), 45- 49.

M

437 Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of

438 bacteriophage T 4. Nature, 227 (5259), 680-685.

D

439 Lan, X. H., Liu, P., Xia, S. Q., Jia, C. S., Mukunzi, D., & Zhang, X. M. (2010). Temperature effect
TE

440 on the non-volatile compounds of Maillard reaction products derived from

441 xylose-soybean peptide system: Further insights into thermal degradation and
EP

442 cross-linking. Food Chemistry, 120(4), 967-972.

443 Liu, A. J., Wei, L. N., Cao D. X., & Liu J. J. (2009). Preparation of barbecue shrimp essence by
C

444 maillard reaction and its analysis by GC-MS. Modern Food Science and Technology,

445 25(6), 674-680.

AC

446 Liu, Q., Kong, B. H., Han, J. C., Sun, C. Y., & Li, P. J. (2014). Structure and antioxidant activity of

447 whey protein isolate conjugated with glucose via the Maillard reaction under dry-heating

448 conditions. Food Structure, 1(2), 145-154.

449 Maillard, M. N., Billaud, C., Chow, Y. N., Ordonaud, C., & Nicolas, J. (2007). Free radical

450 scavenging, inhibition of polyphenoloxidase activity and copper chelating properties of

451 model Maillard systems. Food Science and Technology, 40(8), 1434-1444.

452 Martinez-Alvarenga, M. S., Martinez-Rodriguez, E. Y., Garcia-Amezquita, L. E., Olivas, G. I.,

16
 ACCEPTED MANUSCRIPT

453 Zamudio-Flores, P. B., & Acosta-Muniz, C. H. (2014). Effect of Maillard reaction

454 conditions on the degree of glycation and functional properties of whey protein

455 isolate-Maltodextrin conjugates. Food Hydrocolloids, (38), 110-118.

456 Ogasawara, M., Katsumata, T., & Egi, M. (2006). Taste properties of Maillard-reaction products

457 prepared from 1000 to 5000 Da peptide. Food Chemistry, 99(3), 600-604.

PT
458 Serpen, A., & Gokmen, V. (2009). Evaluation of the Maillard reaction in potato crisps by

459 acrylamide, antioxidant capacity and color. Journal of Food Composition and Analysis,

RI
460 22(6), 589-595.

461 Shimoda, M., & Shibamoto, T. (1990). Isolation and identification of headspace volatiles from

SC
462 brewed coffee with an on-column GC/MS method. Journal of Agricultural and Food

463 Chemistry, 38(3), 802-804.

U
464 Song, N., Tan, C., Huang, M. G., Liu, P., Eric, K., & Zhang*, X. M. (2013). Transglutaminase
AN
465 cross-lingking effect on sensory characteristics and antioxidant activities of Maillard

466 reaction products from soybean protein hydrolysates. Food Chemistry, 136(1), 144-151.
M

467 Su, G., Zheng, L., Cui, C., Yang, B., Ren, J., & Zhao, M. (2011). Characterization of antioxidant

468 activity and volatile compounds of Maillard reaction products derived from different
D

469 peptide fractions of peanut hydrolysate. Food Research International, 44(10),

TE

470 3250-3258.

471 Sun, W. Z., Zhao, M. M., Cui, C., Zhao. Q. Z., & Yang, B. (2010). Effect of Maillard reaction
EP

472 products derived from the hydrolysate of mechanically deboned chicken residue on the

473 antioxidant, textural and sensory properties of Cantonese sausages. Meat Science, 86,
C

474 276-282.

475 Yahya, H., Linforth, R. S. T., & Cook, D. J. (2014). Flavour generation during commercial barley
AC

476 and malt roasting operations: A time course study. Food Chemistry, 145, 378-387.

477 Yu, X. Y., Zhao, M. Y., Hu, J., Zeng, S. T., & Bai, X. L. (2012). Correspondence analysis of

478 antioxidant activity and UV-Vis absorbance of Maillard reaction products as related to

479 reactants. LWT-Food Science and Technology, 46(1), 1-9.

480 Zhang, L. F., & Sui, W. (2006). Study on the heat-reaction shrimp flavors from shrimp. Food and

481 Fermentation Industries, 32(10), 82-85.

482 Zhang, Y., Dorjpalam, B., & Ho, C. T. (1992). Contribution of peptides to volatile formation in

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483 the Maillard reaction of casein hydrolysate with glucose. Journal of Agricultural and

484 Food Chemistry, 40(12), 2467-247.

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18
 Heating pH pH L* L* a* a* b* b* 294 nm 294 nm 420 nm 420 nm

time (min) (TDPs) (MRPs) (TDPs) (MRPs) (TDPs) (MRPs) (TDPs) (MRPs) (TDPs) (MRPs) (TDPs) (MRPs)

0 8.00±0.00a 8.00±0.00a 49.9±1.69a 49.9±1.69a -2.65±0.13c -2.65±0.13c 13.73±0.19a 13.73±0.19a 0.17±0.01a 0.17±0.01f 0.21±0.01b 0.21±0.01ef

10 7.87±0.08abc 7.48±0.10b 47.63±0.65b 46.08±1.48b -2.67±0.07c -2.21±0.10e 12.32±0.14b 14.47±0.16ef 0.14±0.017bc 0.16±0.01f 0.23±0.03a 0.23±0.02ef
AC
20 7.91±0.09ab
C
7.45±0.09b 48.07±0.68b 44.31±1.06b -3.49±0.09b -2.05±0.09e 11.11±0.13c 14.58±0.19e 0.15±0.01b 0.16±0.01f 0.19±0.01bc 0.14±0.01f

30 7.86±0.09abc 7.21±0.03c 47.02±0.98bc 40.46±1.21c -3.75±0.10a -1.91±0.05e 11.05±0.24c 17.01±0.23d 0.14±0.01bc 0.20±0.01f 0.18±0.00bcd 0.19±0.01f
EP
60 7.86±0.08abc 7.05±0.05d 45.42±0.60cd 35.19±0.81d -3.77±0.06a -0.80±0.04d 10.55±0.16d 26.85±0.82b 0.14±0.01bc 0.56±0.02e 0.18±0.01bcd 0.30±0.02e
TE
90 7.75±0.12bc 6.76±0.05e 42.11±1.30e 30.91±0.89e D-3.56±0.08b 8.04±0.26c 9.14±0.11f 28.97±0.58a 0.15±0.01bc 1.04±0.05d 0.17±0.00bcd 0.76±0.05d

120 7.74±0.12c 6.34±0.05f 41.98±0.69e 24.81±0.94f -3.56±0.08b 12.02±0.22a

M 9.26±0.09f 20.55±0.53c 0.15±0.01bc 1.43±0.08c 0.16±0.00bcd 1.2±0.08c

150 7.70±0.08c 6.24±0.05f 41.77±0.85e 21.66±1.31g -3.52±0.06b 12.23±0.32a 9.33±0.14f 15.23±0.54e 0.13±0.01e 1.86±0.06b 0.17±0.01bcd 1.76±0.08b
AN
180 7.72±0.09c 6.04±0.13g 44.01±1.36d 19.58±0.56h -3.73±0.06a 10.19±0.21b 9.92±0.30e 11.82±0.27g 0.14±0.01c 2.19±0.04a 0.17±0.01bcd 2.15±0.11a
ACCEPTED MANUSCRIPT

U SC
RI
PT
Table 1 Development of pH and color formation for MRPs and TDPs as a function of heating time
 ACCEPTED MANUSCRIPT
Table 2
Volatile compounds detected in hydrolysate-xylose systems by heating different time
RT Percentage of total area（%）
No. Compound RI
(min) 30min 90min 150min 180min

Sulphur-containing compounds(3)

1 Dimethyl disulfide 740 4.241 0.4 3.86 3.79 2.09

2 Dimethyl trisulfide 968 11.485 1.69 2.16 4.57 3.42

3 2-Acetylthiazole 1020 13.227 0.43 0.54 0.38 0.53

PT
Nitrogen containing compounds(12)

Pyrazine(5)

4 Pyrazine 672 3.975 0.15

RI
5 Pyrazine, 3-ethyl-2,5-dimethyl- 1082 15.246 0.16 0.17 0.18

6 n-Pentylpyrazine 1178 18.200 0.2

SC
7 Pyrazine, 2,5-dimethyl- 912 9.582 3.14 3.5

8 2,6-dimethyl- pyrazine 911 9.547 0.14 0.18

Pyrroles(1)

U
9 1-(2-furanylmethyl)-1H-Pyrrole 1184 18.361 0.28 0.41 0.51

Diazine(2)
AN
10 1,3-Diazine 721 4.090 0.14

11 Pyrimidine, 4,6-dimethyl- 912 9.570 3.64

Others(4)
M

12 3-Methylpyridazine 670 3.767 0.13 0.14

13 2,4-Quinolinediol 1524 27.394 0.1

14 7H-Dibenzo[c,g]carbazole 1116 16.296 0.15

D

15 Pyridine, 3,4-dimethyl- 1858 34.719 0.16

Furans non-S-containing(4)
TE

16 Furan, 2-pentyl- 991 12.305 2.02 0.68 0.8 0.83

17 2-n-Butyl furan 1417 24.752 0.15

18 Furfural 833 6.952 17.84 29.09 38.2

EP

19 2-Furanmethanol 870 8.174 0.23

Aldehyde compounds(21)

20 3-methyl-butanal 649 2.683 19.36 32.8 23.15 18.63

C

21 Hexanal 799 5.810 6.55 0.69 0.8 0.53

22 Heptanal 903 9.259 4.27 0.3 0.26 0.2

AC

23 Benzaldehyde 960 11.220 3.71 2.15 2.06 2.01

24 Benzeneacetaldehyde 1044 14.023 4.93 14.04 11.26 13.45

25 Benzenepropanal 1164 17.761 0.13 0.16

26 2-Dodecenal, (E)- 1060 14.531 0.82

27 Nonanal 1105 15.973 19.79 2.71 1.87 1.85

28 Decanal 1206 19.030 5.02 1.32 1.2 0.85

29 2-Undecenal 1365 23.402 0.18

30 Pentadecanal- 1817 33.900 0.61 0.77

 ACCEPTED MANUSCRIPT
Table 2 (continued)
RT Percentage of total area（%）
No. Compound RI
(min) 30min 90min 150min 180min

31 Benzaldehyde, 2,3,4-trimethoxy- 1694 31.282 0.89

32 Undecanal 1308 21.880 0.21 0.2

33 Dodecanal 1410 24.556 0.21

34 5-Methyl-2-phenyl-2-hexenal 1493 26.644 0.09

35 Benzeneacetaldehyde, .alpha.-ethylidene- 1275 20.957 0.05

PT
36 Benzaldehyde, 3,4-dimethyl- 1217 19.318 0.06

37 Methional 907 9.397 0.76 0.54

38 2,6-Nonadienal, (E,E)- 1155 17.484 0.03

RI
39 Butanal 589 2.014 0.83

40 2-Decenal, (E)- 1365 23.402 0.05

SC
Ketone compounds(12)

41 2,3-Octanedione 951 10.920 0.38 1.24 0.27

42 5-Hepten-2-one, 6-methyl- 988 12.201 1.2 0.35 0.37 0.29

U
43 Acetophenone 1067 14.750 0.17 0.18 0.21

44 4-Acetyl-1-methylcyclohexene 1133 16.815 0.09

AN
45 2-Undecanone 1295 21.522 0.23 0.12

46 2-Nonanone 1093 15.604 0.1 0.15

47 2-Hexanoylfuran 1268 20.772 0.08

M

48 4-Nonanone 951 10.932 1.13

59 1-Pentanone, 1-phenyl- 1067 14.750 0.27

50 2-Nonadecanone 1294 21.510 0.17

D

51 5,9-Undecadien-2-one, 6,10-dimethyl-, (Z)- 1455 25.687 0.36

52 3-Benzylidene-2,4-pentanedione 1534 27.625 0.08

TE

Alcohol compounds(11)

53 1-Octen-3-ol 982 11.981 0.71

54 2-Decen-1-ol, (E)- 1070 14.865 0.29 0.12

EP

55 3-Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)-, (R)- 1136 16.930 0.28

56 L-.alpha.-Terpineol 1193 18.626 0.64 0.33 0.26

57 1-Hexanol, 2-ethyl- 1031 13.608 0.52

C

58 1-Triacontanol 1612 29.436 0.14

59 4-Ethylcyclohexanol 1278 21.060 0.34

AC

60 1-Decanol, 2-hexyl- 1577 28.628 1.51

61 Benzenemethanol, 4-methoxy- 1345 22.872 0.55

62 1-Nonen-3-ol 982 11.970 0.21

63 1-Hexanol, 2-ethyl- 1031 13.596 0.4

Ester compounds(3)

64 Formic acid, octyl ester 1073 14.969 1.43

65 Isopropyl myristate 1827 34.085 15 5.94 2.74 1.82

66 Hexadecanoic acid, methyl ester 1927 36.092 0.15 0.08

 ACCEPTED MANUSCRIPT
Table 2 (continued)
RT Percentage of total area（%）
No. Compound RI
(min) 30min 90min 150min 180min

Acid compounds(4)

67 n-Hexadecanoic acid 1962 36.761 0.06

68 Cystine 1373 23.610 0.01 0.04

69 Benzeneacetic acid 1090 15.489 0.11

70 2-Octenoic acid 1373 23.610 0.04

PT
Hydrocarbon compounds(23)

Alkane(18)

71 2,3-Epoxybutane 540 2.164 3.76

RI
72 Benzene, 1-methyl-3-(1-methylethyl)- 1025 13.389 0.18

73 Cycloheptane 1174 18.061 0.21

SC
74 Naphthalene 1183 18.349 0.36

75 Naphthalene, 2-methyl- 1311 21.972 0.06 0.02

76 Nonadecane 1399 24.302 0.12

U
77 Naphthalene, 1,4-dimethyl- 1421 24.844 0.09

78 Eicosane 1799 33.531 1.03 1.01 0.65

AN
79 Heptadecane 1699 31.397 0.41 0.94

80 Octadecane 1799 33.531 0.2 0.56 0.28

81 Naphthalene, 2,6-dimethyl- 1425 24.937 0.05

M

82 Naphthalene, 2,7-dimethyl- 1421 24.845 0.05

83 o-Cymene 1024 13.377 0.12

84 Naphthalene, 1-methyl- 1312 21.983 0.22 0.03

D

85 Oxirane, hexadecyl- 1920 35.954 0.27

86 Dodecane 1549 27.971 0.05

TE

87 Cyclopentadecane 1552 28.051 0.13

88 Pentadecane 1900 35.562 0.1

Alkene(5)
EP

89 D-Limonene 1028 13.516 0.82 0.24 0.89 0.3

90 Naphthalene, 2-ethenyl- 1486 26.471 0.12 0.07

91 Hexadecane 1599 29.159 2 0.56 1.64 0.63

C

92 Cyclohexene, 1-methyl-4-(1-methylethyl)-, (R)- 1075 15.027 0.52

93 1-Tridecene 1296 21.567 0.06

AC

Amine compounds(3)

94 Benzenemethanamine, .alpha.,4-dimethyl- 1132 16.792 0.1

95 1,4-Butanediamine 760 4.392 0.11 0.03

96 o-Toluidine, 5-isopropyl- 1396 24.221 0.07

Phenols compounds(4)

97 1-Naphthalenol 1246 20.161 0.07 0.11

98 Creosol 1086 15.385 0.19

99 Phenol, 2,6-bis(1,1-dimethylethyl)- 1442 25.375 0.06 0.1 0.07

100 Phenol, 2,4-bis(1,1-dimethylethyl)- 1515 27.163 4.33 2.84 2.42

 Thermal degradation time (min) Maillard reaction time (min)
Free amino acid
0 30 60 120 180 0 30 60 120 180
of heating time.

Asp 10.27 ±0.04a 10.00 ±0.03b 7.54 ±0.02e 7.78 ±0.02d 7.94 ±0.03c 10.27 ±0.31a 5.84 ±0.12c 7.60 ±0.04b 7.70 ±0.06b 7.72 ±0.02b

Thr 22.41± 0.03a 14.99 ±0.08b 8.08 ±0.02c 7.63 ±0.03d 7.60 ±0.01d 22.41 ±0.24a 8.67 ±0.12b 8.48 ±0.05b 7.13 ±0.08c 6.90 ±0.01c
AC
Ser 9.78 ±0.02a 8.93 ±0.04b 5.83 ±0.02e 6.63 ±0.03d 6.86 ±0.02c 9.78 ±0.31a 5.28 ±0.07d 5.72 ±0.07c 6.13 ±0.04b 6.13 ±0.00b
C
Glu 19.16 ±0.02a 17.55 ±0.03b 10.50 ±0.02e 14.97 ±0.04d 16.37 ±0.06c 19.16 ±0.23a 11.38 ±0.13d 10.70 ±0.04e 14.33 ±0.06b 14.08 ±0.04c

Gly 15.55 ±0.04a 13.92 ±0.04b 9.52 ±0.02e 9.63 ±0.03d 9.80 ±0.03c 15.55 ±0.38a 7.44 ±0.16d 9.35 ±0.08b 8.64 ±0.07c 8.32 ±0.03c
EP
Ala 22.56 ±0.03a 19.90 ±0.08b 13.22 ±0.06e 13.44± 0.04d 13.61 ±0.05c 22.56 ±0.25a 9.95 ±0.35d 13.20 ±0.08b 12.37 ±0.06c 12.10 ±0.01c

Cys 13.07± 0.05a 12.86 ±0.05b 5.96 ±0.02c 5.16 ±0.03d 5.04± 0.02e 13.07 ±0.20a 5.31 ±0.14c 5.67 ±0.06b 5.43 ±0.02c 4.73 ±0.05d
TE
Val 17.75 ±0.03a 16.39 ±0.04b 12.58 ±0.05e 12.65 ±0.04d 12.69 ±0.03c 17.75 ±0.18a 10.77 ±0.22d 12.74 ±0.09b 12.28 ±0.08c 12.12 ±0.03c
D
Met 11.29 ±0.03a 10.25 ±0.03b 7.97 ±0.03c 7.79 ±0.03d 7.67 ±0.03e
M 11.29 ±0.22a 6.73 ±0.17d 7.80 ±0.08b 7.63 ±0.02bc 7.41 ±0.02c

Ile 14.18 ±0.06a 12.58 ±0.03b 8.66 ±0.02c 8.58 ±0.02d 8.49 ±0.03e 14.18 ±0.29a 6.72 ±0.08d 8.70 ±0.05b 8.21 ±0.03c 7.99 ±0.02c

Leu 26.24 ±0.04a 23.24 ±0.06b 16.11 ±0.04c 15.98 ±0.05d 15.83± 0.04e 26.24 ±0.15a 12.37 ±0.15e 15.92 ±0.10b 15.07 ±0.03c 14.73 ±0.04d
AN
Tyr 21.81± 0.05a 20.20 ±0.05b 16.48± 0.06c 15.89 ±0.03d 15.80 ±0.02e 21.81 ±0.30a 15.50 ±0.16e 17.73 ±0.07b 16.75 ±0.05c 16.34 ±0.02d
ACCEPTED MANUSCRIPT

Phe 21.63 ±0.04a 19.47 ±0.06b 15.76 ±0.04c 15.11 ±0.03d 15.08± 0.06d 21.63 ±0.41a 13.69 ±0.11d 15.67 ±0.04b 15.07 ±0.10c 14.95 ±0.05c
U
Lys 24.21 ±0.03a 22.26 ±0.04b 18.37 ±0.02c 17.69 ±0.04d 17.58 ±0.06e 24.21 ±0.07a 15.47 ±0.22e 17.54 ±0.08b 16.27 ±0.06c 15.94 ±0.02d

His 6.07 ±0.03a 5.47 ±0.03c 6.00 ±0.02b 5.44 ±0.02c 5.36 ±0.01d 6.07 ±0.13a 5.55 ±0.09c 5.76 ±0.04b 5.30 ±0.02d 5.14 ±0.01e
SC
Arg 35.83 ±0.06a 32.41 ±0.04b 22.26 ±0.06d 22.35 ±0.03d 23.17 ±0.05c 35.83 ±0.44a 16.80 ±0.07c 19.55 ±0.09b 16.20 ±0.06d 14.67 ±0.03e

Pro 33.96 ±0.07a 31.39 ±0.06b 24.82 ±0.07d 25.85 ±0.06c 25.97 ±0.08c 33.96 ±0.40a 25.14 ±0.06c 29.62 ±0.11b 25.17 ±0.06c 25.24 ±0.03c
RI
Total 325.76± 0.31a 291.80± 0.23b 209.66 ±0.08e 212.55± 0.14d 214.85 ±0.20c 325.78 ±0.70a 182.62 ±0.80e 211.76 ±0.42b 199.68 ±0.64c 194.50 ±0.04d
PT
Table 3 Changes in free amino acids content (mg/g) of TDPs and MRPs for CSWHs as a function
 ACCEPTED MANUSCRIPT

a b
control control
7 7
度80 min 6 0 min 度80 min 6 0 min
5 5
4 4
3 meaty
3
度50 min 强 度0 min seafood 度50 min 度0 min meaty
强
度 fis长y 度 seafood
0 toasty 0 fis长y
burnt toasty
度强0 min 强0 min 度强0 min 强0 min burnt

90 min 30 min

PT
90 min 30 min
60 min
60 min

RI
c
control control
7 7
度80 min 6 0 min 度80 min 6 0 min

SC
5 5
umami 4 umami
4
3 continuity 3 continuity
度50 min 强 度0 min bitterness 度50 min 强 度0 min bitterness
度 saltiness
度
0 saltiness
0
sourness sourness
度强0 min 强0 min mout长fulness 度强0 min 强0 min mout长fulness

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sweetness sweetness

90 min 30 min 90 min 30 min

AN
60 min 60 min

Fig. 1. Aroma sensory profiles of TDPs (a) and MRPs (b) as a function of heating time. Taste
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sensory profiles of TDPs (c) and MRPs (d) as a function of heating time. Each value represents the
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average taste intensity scored on a scale ranging from 0 to 7.

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a b
0min 0min
1000 10min 1000 10min
20min 20min
800 30min 800 30min
60min 60min
90min 90min
600 120min 600
120min
150min 150min
400 180min 400 180min

200 200

0 0

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350 400 450 500 550 350 400 450 500 550

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c d
3.5 3.5
0min 180min
10min 150min
3.0 3.0
20min 120min
30min 90min
2.5 2.5

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60min 60min
90min 30min
2.0 A 2.0 20min
120min
A

150min 10min
1.5 180min 1.5 0min

1.0 1.0

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0.5 0.5

0.0 0.0
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200 250 300 350 400 450 500 550 600 650 700 200 250 300 350 400 450 500 550 600 650 700
λ(nm) λ nm

Fig. 2. The emission fluorescence spectra and UV-Vis spectra of the TD (a, c) and MR (b, d)
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model system as a function of heating time.

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M 0min 30min 60min 90min 120min 150min 180min M 0min 30min 60min 90min 120min 150min 180min
a b

66KD 66KD
45KD 45KD
35KD 35KD
27KD 27KD

20KD 20KD

14.4KD
14.4KD
9.5KD
9.5KD
6.5KD 6.5KD

PT
4.1KD 4.1KD

Fig. 3 The SDS-PAGE patterns of hydrolysates (a) and hydrolysates-xylose (b) model systems

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formed with increasing heating time

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Highlights
The utilization of Chinese shrimp waste reduce environment pollution.
Chinese shrimp waste hydrolysates MRPs has strong enticing flavour and colour.
Reaction time plays important role in the generation of flavour compounds for MRPs.

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U SC
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