Accepted Manuscript

Production of a minimally processed jelly candy for children using honey instead of
sugar

Ceren Mutlu, Sultan Arslan Tontul, Mustafa Erbaş

PII: S0023-6438(18)30288-3
DOI: 10.1016/j.lwt.2018.03.064
Reference: YFSTL 6993

To appear in: LWT - Food Science and Technology

Received Date: 7 February 2017

Revised Date: 21 March 2018
Accepted Date: 24 March 2018

Please cite this article as: Mutlu, C., Tontul, S.A., Erbaş, M., Production of a minimally processed jelly
candy for children using honey instead of sugar, LWT - Food Science and Technology (2018), doi:
10.1016/j.lwt.2018.03.064.

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 ACCEPTED MANUSCRIPT
1 Production of a minimally processed jelly candy for children using honey instead

2 of sugar

4 Ceren MUTLU1,2 Sultan ARSLAN TONTUL1,3 Mustafa ERBAŞ1*

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6 Department of Food Engineering, Faculty of Engineering, Akdeniz University, 07058

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7 Antalya, Turkey
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8 Department of Food Engineering, Faculty of Engineering, Balikesir University, 10145

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9 Balikesir, Turkey
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10 Department of Food Engineering, Faculty of Agriculture, Selcuk University,

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42130 Konya, Turkey
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12 *Corresponding author: Tel: +90 242 310 6575; Fax: +90 242 310 6306
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13 e-mail: erbas@akdeniz.edu.tr

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26 Abstract

27 This study investigated some chemical, physical and sensorial properties of honey jelly

28 candies made by two different mixing techniques (cold and hot), three gelatine doses (15, 20

29 and 25% of honey weight) and three fruit juices (orange, strawberry and black mulberry).

30 The mean water content and activity of samples were 23.38% and 0.73, respectively.

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31 The acidity and diastase number of samples ranged between 40.12-46.09 meq/kg and 15.43-

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32 0.00, respectively. While diastase activity of honey was preserved more than 95% by cold

33 mixing technique in candy form, it was completely lost in the hot mixing technique.

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34 Increasing of gelatine dose, increased the hardness, adhesiveness, chewiness and gumminess

35 values of candies. Additionally, the glucose, fructose and sucrose contents of cold mixed

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honey jelly candies were determined as 403.23±5.24, 491.10±7.61 and 1.38±0.12 g/kg,
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37 respectively. Addition of fruit juices increased the acidity value and sugar and proline
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38 contents of samples. The sensorial acceptability score for honey jelly candies obtained more

39 than 3.5 on a 5-point hedonic scale.

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40 In conclusion, a new jelly candy as a healthier alternative to the existing ones was
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41 produced by minimal processing with honey and fruit juices instead of sugar syrup and

42 artificial additives without inactivation of bioactive compounds of honey.

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43
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44 Keywords: Honey, Diastase Enzymes, Gelatine, Candy, Children

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51 1. Introduction

52 A high rate of candy consumption can negatively impact on public health, especially

53 children’s health, because of the contaminants (5-hydroxymethyl-2-furaldehyde, acrylamide,

54 etc.) occurring with heat treatment in the production process. Additionally, the ingredients

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55 such as sugar and artificial food additives (flavouring and colouring agents) also have

56 negative effects on health. It is estimated that the amount of candy consumption is

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57 approximately 7 kg per person in high-income countries and whilst it is much lower in the

58 low-income countries, this market is growing daily (Palacıoglu, 2003). Different kinds of

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59 confectionery products such as soft and jelly candy, croquant, and nougat are produced, and

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60 the gummy jelly candy group account for about 50% of confectionery market (Garcia, 2000).
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61 Gummy jelly candies are produced by boiling of gelling agents and sugars at high

62 temperatures (>100°C). After boiling, the flavouring and colouring agents are added into the
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63 mixture (Riedel, Bohme, & Rohm, 2015). The obtained mixture is moulded in dried corn

64 starch moulds and they are kept at 65°C for 12 hours, and after moulding step they are cut out
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65 (Habilla, & Cheng, 2015). In some studies; the gelatine, Ƙ-carrageenan, pectin, guar and
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66 xanthan gums, starch and their derivatives are used as gelling agent for providing gel structure
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67 in the jelly candy production (Charoen, Savedboworn, Phuditcharnchnakun, & Khuntaweetap,

68 2015; Habilla & Cheng, 2015; Utomo, Darmawan, Hakim, & Ardi, 2014). However, the main
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69 gelling agent of these candies is gelatine (Charoen, Savedboworn, Phuditcharnchnakun, &

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70 Khuntaweetap, 2015). Additionally, glucose and fructose syrups, dextrose and sucrose are

71 used as sugar source in the candy formulation (Charoen, Savedboworn, Phuditcharnchnakun,

72 & Khuntaweetap, 2015; Ergun, Lietha, & Hartel, 2010; Habilla & Cheng, 2015; Utomo,

73 Darmawan, Hakim, & Ardi, 2014).

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75 A healthy alternative to sugar is honey. Honey is a functional food produced by bees

76 (Apis mellifera) which contains a wide variety of bioactive substances and enzymes giving it

77 antioxidant, antimicrobial, anti-inflammatory and anticarcinogenic properties (Eteraf-Oskouei

78 & Najafi, 2013; Samarghandian, Farkhondeh, & Samini, 2017), although it has high sugar

79 content (Karabagias, Badeka, Kontakos, Karabournioti, & Kontominas, 2014). It is composed

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80 of carbohydrates (82%), water (17%), proteins (0.3%), minerals (0.7%), vitamins, organic

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81 acids, enzymes, phenolic acids and flavonoids (Islam, Khalil, Islam, Moniruzzaman, Mottalib,

82 Sulaiman, et al., 2012), with the major carbohydrates being fructose (38%), glucose (31%)

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83 and sucrose (<8%) (Ball, 2007; Hussein, Yusoff, Makpol, & Mohd, 2014).

84 There are two types of honey based on the source of nectar used by the bees, blossom

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and honeydew. The nectar used to produce blossom honey is from flowers such as thyme,
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86 clover, acacia and citrus, while the nectar used for honeydew honey is from herbs or insects
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87 living on herbs, for example, pine, oak and fir honey (Karabagias, Badeka, Kontakos,

88 Karabournioti, & Kontominas, 2014).

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89 The aim of this research was to produce a minimally processed jelly candy with honey
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90 and fresh fruit juices instead of sugar syrup and artificial food additives without inactivation

91 of their bioactive compounds, thereby providing a healthier candy product.

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92

2. Material and methods

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93
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94 2.1. Materials

95 Blossom honey and fruits (orange, strawberry and black mulberry) were purchased from

96 well-known local markets in Antalya, Turkey and bovine gelatine was obtained from Ewald-

97 Gelatine (GmbH, Sobernheim, Germany). Analytical and chromatographical grade chemicals

98 were obtained from Merck (Darmstadt, Germany) and Sigma (Taufkirchen, Germany).

99
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100 2.2. Experimental design

101 This research consisted of two parts. As the first part of this study, the effects of two

102 mixing techniques (cold and hot) and three gelatine doses (15, 20 and 25% of honey weight)

103 on properties of honey jelly candy were investigated. A sensorial evaluation was carried out

104 with produced honey jelly candies to determine the best mixing technique and gelatine dose to

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105 produce honey jelly candies with fruit juices. In the second part of study, the chosen honey

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106 jelly candy formulation from sensorial test was referred as plain control sample and re-

107 produced by adding different flavours and colours by using fresh orange, strawberry and

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108 black mulberry juices. The first (n; 2x3x2=12) and second (n; 4x2=8) stages were performed

109 in two replicates.

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111 2.3. Production of honey jelly candies
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112 In the first part of the study, candies were produced using two different mixing

113 techniques; cold (50-55°C) and hot (115°C), with three different gelatine doses; 15%, 20%
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114 and 25% of honey weight. Each gelatine dose was weighted into a beaker with 50 mL volume
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115 and the beaker was filled by adding distilled water. The gelatine was kept inside water for 5

116 min and the excess water completely removed. It was determined that gelatine absorbed water
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117 at 1.25 fold its weight during soaking. This soaked gelatine was used as gelling agent in the

two different mixing techniques.

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119 In the cold mixing technique; the soaked gelatine in the beaker was dissolved at 70°C

120 for 30 min in a water bath (Memmert Waterbath WNE 29, Schwabach, Memmert, Germany).

121 Dissolved gelatine was added to the honey (25°C) and then mixed at 200 rpm for 1 min with a

122 stirrer (Wisestir HS-30D, Daihan Scientific, Korea). There was no heat treatment during

123 mixing, but the temperature of mixture was measured at room temperature (25°C) between

124 50-55°C because of pre-heat treatment of gelatine dissolved at 70°C in the water bath.
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125 In the hot mixing technique, candies were produced according to conventional

126 confectionery production. For this purpose, the soaked gelatine was added into the weighted

127 honey and they were manually stirred by glass rod on a hot plate for 20 min. During this

128 process, the temperature of honey-gelatine mixture was measured at approximately 115°C.

129 In the both techniques, honey and gelatine mixtures were moulded into a heart shape

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130 silicone mould (13 x 15 x10 mm) and kept for 30 min in a fridge at 4°C. The honey jelly

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131 candies were then unmoulded and kept for three days in a desiccator containing saturated

132 potassium carbonate solution (43.2% relative humidity). After this process, the honey jelly

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133 candies were placed in plastic bags and stored at +4°C.

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135
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2.4. Production of fruit juice added honey jelly candies
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136 In the second part of the study, honey jelly candies with fresh fruit juices were
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137 produced by using orange, strawberry and black mulberry juices instead of distilled water.

138 The fruity honey jelly candy production technique and gelatine dose were decided according
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139 to results of the sensorial evaluation conducted with the plain honey jelly candies produced in
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140 the first part of this study (Section 2.3). The fruits were obtained daily in a local market and

141 smashed in a blender and their juices were filtered. The obtained fruit juices were kept at
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142 +4°C.

The gelatine, selected as 15% in sensorial evaluation, was weighted into a beaker with
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144 50 mL volume and the beaker was filled with fruit juice and it was kept for 5 min. After

145 removing excess fruit juice, the soaked gelatine was dissolved at 70°C for 30 min in the water

146 bath. Dissolved gelatine was added to the honey and then mixed at 200 rpm for 1 min with a

147 stirrer. The mixture was moulded and stored as describing in the Section 2.3. The plain honey

148 jelly candy (cold mixed with 15% gelatine) was used as a control and produced with distilled

149 water using the same conditions.

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150

151 2.4. Determination of moisture content and water activity

152 The moisture content was determined by drying 1 g of the sample in a vacuum oven at

153 60°C (Memmert VO200, Schwabach, Germany) to a constant weight (Periche, Castello,

154 Heredia, & Escriche, 2016) and the water activity (aw) of the honey jelly candies was

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155 measured using an Aqua Lab 4TE (Decagon Devices, USA) water activity meter.

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156

157 2.5. Determination of pH value and titratable acidity

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158 The pH value of honey jelly candies was determined by homogenising 1 g of sample in

159 9 mL distilled water (50°C) with an ultraturrax (Ultraturrax T-25, IKA Labortechnik, Staufen,

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Germany). The pH of suspension was measured using a digital pH meter (WTW 537,
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161 Weilheim, Germany).
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162 The titratable acidity of sample was determined by titrating the prepared suspension as

163 described in the pH analysis with 0.1N NaOH up to pH 8.1, while mixing continually on
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164 magnetic stirrer and the results were expressed as meq/kg (Bhat, Kamaruddin, Min-Tze, &
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165 Karim, 2011).

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167 2.6. Determination of diastase number

The diastase number of honey and candies was determined with a spectrophotometric
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169 method. For this purpose, 2 g sample was homogenised with 1 mL acetate buffer solution

170 (1.59 M) and 4 mL distilled water using the ultraturrax for 2 min. After that, 0.6 ml NaCl

171 solution (0.5 N) was added to 2 mL of the obtained solution and total volume was adjusted to

172 10 mL with distilled water.

173 Adjustment of initial absorbance of starch solution to 0.745–0.770 nm was carried out

174 to determine the amount of water to add into the sample. For this purpose, 5 mL of starch
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175 solution (2%) was mixed with 10 mL distilled water and 0.5 mL of this solution was then

176 added to ten different beakers containing 5 mL 0.0007 N iodine solution and different

177 volumes of water (10, 12, 14, 16, 18, 20, 21, 22, 23 and 24 mL). Absorbance was determined

178 at 660 nm by a spectrophotometer (Shimadzu UV-1800, Kyoto, Japan) and the amount of

179 water in the beaker, which has absorbance value between 0.745-0.770, should be added to the

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180 sample before the absorbance measurement.

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181 Prepared sample solutions (2 mL) and starch solution (2%) were placed in the water

182 bath at 40°C for 15 min before the addition of 1 mL of starch solution. Then, 0.1 mL of the

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183 prepared solutions, the specified volume of water (1/5) as described above and 1 mL of iodine

184 solution were mixed and the absorbance was determined in every 5 min. The analysis was

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continued until the absorbance values reached 0.235 and the measured absorbance values
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186 were plotted against time to determine the time to achieve an absorbance of 0.235. The
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187 diastase numbers of the samples were calculated as follows:

188 Diastase number = 300/t0.235

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189 t0.235 represents the determined time coincidence to 0.235 from the plot (Bogdanov,
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190 Martin, & Lullmann, 2002).

191
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192 2.7. Determination of sugar profile and proline content

The sugar profile and proline content of honey jelly candies were determined using a
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194 GC-MS system. For sample extraction, 1 g honey jelly candy was suspended in 20 mL ultra-

195 pure water in a centrifuge tube, homogenized at 11000 rpm with the ultraturrax for 1 min

196 prior to centrifugation (Centrifuge, 3-18K, Sigma, Germany) at 12000 rpm for 10 min. The

197 supernatant was then filtered through 0.45 µm filter (Milipore, Billerica, USA) and 50 µL of

198 the solution was evaporated under N2 flow in a vial. For derivatization, 500 µL oximat reagent

199 (hydroxylamine hydrochloride 30 mg/mL dissolved in pyridine) was added and incubated at
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200 70°C for 30 min before the addition of 500 µL of silylation reagent (N,O-

201 bis(trimethylsilyl)trifluoroacetamide, BSTFA) containing 1% trimethylchlorosilane (TMCS).

202 The derivatised aliquot was injected (2 µL) into the GC-MS system (Agilent 7890A

203 GC, 5975C MS, Santa Clara, USA) with 1:10 split ratio. A non-polar column (Agilent HP-1,

204 100% dimethylpolysiloxane, 25 m x 0.32 mm x 1.05 µm, Wilmington, DE, USA) was used

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205 and the helium carrier gas flow rate was adjusted to 1 mL/min. The initial oven temperature

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206 was set to 80°C, increased at 4°C/min to 210°C and later, 2°C/min to 250°C. After that, the

207 final temperature was set at 4°C/min to 280°C and held for 10 min at this temperature. Also,

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208 the temperatures of inlet and MS detector were set to 280°C and 230°C, respectively.

209 Sugars (glucose, fructose and sucrose) and proline standards were used for

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determination of retention times and mass-fragmentation spectra. Standards were dissolved in
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211 water and derivatised using the same conditions described above for samples. The sugar and
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212 proline peaks were determined with external standards and main ion fragments (glucose: 73,

213 103, 147, 205, 319 (m/z); fructose: 73, 103, 147, 217, 307; sucrose: 73, 103, 147, 217, 361,
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214 437; proline: 73, 142, 216, 257) (Cocchi, Durante, Grandi, Lambertini, Manzini, & Marchetti,
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215 2006).

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217 2.8. Determination of colour

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218 The colour parameters (L*, a* and b*) of honey jelly candies were measured by the
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219 CIELAB system with using a CR-400 Chroma Meter (Konica Minolta, Tokio, Japan). The L*

220 colour parameter represents the dark-light spectrum with a range of black (0) to white (100).

221 The a* colour parameter represents red–green colour range with positive values indicating the

222 redness and negative values indicating the greenness. The b* colour parameter represents

223 yellow–blue colour range with positive values indicate yellowness and negative values

224 indicate blueness (Kurek, Wyrwisz, Piwińska, & Wierzbicka, 2015).

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225

226 2.9. Determination of texture profile

227 The texture analysis of honey jelly candies was performed using a texture analysis

228 device (TA TX Plus; Stable Micro Systems, Surrey, UK) equipped with an SMS5 cylinder

229 probe (35 mm). The analysis was performed at room temperature and texture analysis device

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230 parameters were pre-test speed of 2 mm/s, test speed of 1 mm/s, post-test speed of 1 mm/s,

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231 distance between probe and sample of 10 mm, trigger force of 5 g and the delay between two

232 compressions was 2 seconds (Khouryieh, Aramouni, & Herald, 2005).

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233

234 2.10. Sensorial analysis

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In the first part of research, the honey jelly candies were evaluated with respect to
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236 mixing techniques (cold and hot) and gelatine dose (15%, 20% and 25% of honey weight). As
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237 a second part of the research, selected plain sample (produced by cold mixing technique and

238 15% gelatine) was used as control and it was compared with jelly candy samples produced
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239 with different fruit juices (orange, strawberry and black mulberry). The honey jelly candies
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240 were placed on white plastic plates with a glass of water, coded and served to panellists at

241 random order on a white bench under the light. The samples were evaluated by eight trained
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242 panellists who were research assistants in the Department of Food Engineering of Akdeniz

University, Turkey. The descriptive terms selected were appearance, texture, taste and overall.
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243
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244 The acceptability of the honey jelly candies was scored by the panellists on a 5-point hedonic

245 scale (1: disliked extremely, 5: liked extremely) (Meilgaard, Carr, & Civille, 2006).

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247 2.11. Statistical analysis

248 All analyses were performed in duplicate. Statistical data analysis was performed using

249 SAS software statistical software package (v.7.00, SAS Institute Inc., Cary, NC, USA) and
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250 the significance of differences between the selected parameters was tested by one-way

251 ANOVA. The results were given as the mean ± standard error. Each mixing type was used at

252 each different dose level and vice versa. The chemical results were calculated on dry base.

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254 3. Results and discussion

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255 3.1. Honey jelly candies

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256 3.1.1. The effects of mixing technique and gelatine dose on the chemical properties of the

257 candies

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258 Some physicochemical properties of honey jelly candies are given in Table 1. The

259 moisture content and water activity of honey jelly candies were not significantly (p>0.05)

260
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affected by the mixing technique and gelatine dose, and they were 23.38% and 0.73,
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261 respectively. It is considered that the honey jelly candy mixtures were not vaporized enough
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262 during the hot mixing because of the colligative effect of the high sugar concentration and

263 water absorption capacity of the gelatine. Additionally, the water content and activity of
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264 honey jelly candies produced by the cold mixing technique were similar to samples produced
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265 by hot mixing technique due to the three days in a desiccator (43.2% RH). A previous study

266 reported a water content and activity of jelly candies as 18-22% and 0.50-0.75, respectively
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267 (Bussiere & Serpelloni, 1985). In another study, the water activity of jelly candies was

reported as 0.74-0.77 (Permata & Sayuti, 2014).

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268
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269 The titratable acidity and pH value of honey jelly candies were significantly affected

270 (p<0.01; p<0.05) by the mixing technique and gelatine dose. It was determined that the hot

271 mixing technique (115°C) increased the total titratable acidity and decreased the pH value of

272 honey jelly candies. The alterations in acidity and pH in honey jelly candies may be due to the

273 sugar acids formed by the oxidation of hexose at the high temperature, as it is known that

274 sugars convert to sugar acids in weak acid media at a high temperature (Fennema, 1996). As
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275 predicted, the acidity decreased with increasing the gelatine dose because of the decreasing

276 ratio of honey in the mixture. There are previous reports of pH value of candies ranging from

277 3.35 to 3.39 (Khouryieh, Aramouni, & Herald, 2005) and between 3.65-4.29 (Ventura et al.,

278 2013).

279 The diastase number is determined by the activity of diastase enzymes and it decreases

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280 with increasing temperature, processing and storage time (Subramanian, Umesh Hebbar, &

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281 Rastogi, 2007; Tosi, Martinet, Ortega, Lucero, & Ré, 2008). This research was performed

282 with honey which has a diastase number of 16.10 and this value was within legal limits

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283 according to Codex Alimentarius Standard for honey (codex stan: 12-1981) (Codex

284 Alimentarius, 2001). The diastase numbers of 45 honey samples obtained from Turkey were

285
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reported as 8.7-26.1 for fresh honey and 6.7-18.2 for one-year stored honey (Yilmaz &
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286 Küfrevioğlu, 2001). Also, in another study, the diastase number of honey samples was
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287 determined between 1.47-49.42 (Serrano, Villarejo, Espejo, & Jodral, 2004).

288 The diastase activity of honey jelly candies was significantly (p<0.01) affected by the
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289 mixing technique, but there was no significant (p>0.05) difference between gelatine dose. The
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290 diastase number of honey jelly candies produced with a hot mixing technique was zero

291 because diastase enzymes were inactivated by high temperature (115°C). It was reported that
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292 the amylase activity of honey reduced between 2-5 units when honey was heat treated at 85°C

(Babacan, Pivarnik, & Rand, 2002). The mean diastase number was determined as 15.43 in
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293
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294 candies produced by the cold mixing technique (55°C), indicating that the cold mixing

295 technique preserved the diastase enzyme activity of honey after candy production.

296 The sugar profile and proline content of honey jelly candies are presented in Table 2.

297 The glucose, fructose and sucrose contents of honey jelly candies were significantly (p<0.01)

298 affected by the mixing technique and gelatine dose. Glucose, fructose and sucrose content of

299 samples produced with cold mixing technique were higher than the hot mixed samples. The
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300 decreasing of glucose and fructose may be as a result of Maillard reactions during heat

301 treatment. Additionally, sucrose is not an invert sugar, therefore it cannot interact with amino

302 groups during Maillard reactions (Schebor, Burin, Buera, & Chirife, 1999). The decreasing

303 sucrose may be sourced from the chemical inversion of sucrose to glucose and fructose. Also,

304 it was determined that the sugar content of candies decreased with the increasing gelatine

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305 dose, possibly due to the dilution of honey with the increasing gelatine dose in the mixture.

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306 The proline content of honey jelly candies was significantly (p<0.01) affected by the

307 different mixing technique and gelatine dose. The proline content of candies produced by the

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308 cold mixing technique was higher than the hot mixing technique. This may be as a result of

309 proline being used in Maillard reactions during heat treatment. The amount of proline

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decreased with increasing gelatine dose due to dilution of honey in the candy mixture.
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311
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312 3.1.2. The effects of the mixing technique and gelatine dose on the physical properties of the

313 candies
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314 The colour analysis results are presented in Table 3. It was determined that L*, a* and
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315 b* colour parameters of jelly candies were significantly affected (p<0.01; p<0.05) by the

316 mixing technique, but they were not significantly (p>0.05) affected by the gelatine dose.
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317 The L*, a* and b* values indicated that the colour of the honey jelly candies got darker

during the hot mixing technique. This may be as a result of non-enzymatic browning
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318
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319 reactions, such as the Maillard reaction and caramelisation at high temperatures. The Maillard

320 reaction occurs among amino groups and reducing sugars in honey, with high temperatures

321 accelerating such reactions resulting in changes in the aroma, taste and colour of foods

322 (Martins, Jongen, & Van Boekel, 2000).

323 The texture profile analysis results of honey jelly candies are given in Table 4. Texture

324 profile analysis was composed of hardness, adhesiveness, cohesiveness, springiness,

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325 chewiness and gumminess properties. It was determined that texture parameters were not

326 significantly (p>0.05) affected by the different mixing technique except for adhesiveness and

327 springiness. Adhesiveness is the stickiness between product and a surface and related to the

328 product’s molecular structure. The hot mixing technique resulted in a stronger intermolecular

329 force because of conglutination of the ingredients at a high temperature. Subsequently, the

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330 adhesiveness values of candies produced by hot mixing were lower than the cold produced

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331 candies. A previous study reported the cohesiveness and springiness of different jelly candies

332 were between 0.54-0.82 and 0.94-1.49, respectively (Khouryieh, Aramouni, & Herald, 2005).

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333 It was determined that texture parameters were significantly (p<0.01; p<0.05) affected

334 by the gelatine dose, except for cohesiveness. The increasing of gelatine dose increased

335
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hardness, adhesiveness, chewiness and gumminess, but decreased the springiness. The
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336 hardness of candies increased due to the formation of a harder gel structure with the
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337 increasing gelatine dose. Chewiness and gumminess also increased as they are determined by

338 the degree of hardness. Furthermore, the increasing hardness decreased the springiness of the
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339 candies.
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340

341 3.1.3. The effects of the mixing technique and gelatine dose on sensorial properties of candies
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342 Sensory analysis results of honey jelly candies are presented in Figure 1. It was

determined that sensory analysis results were not significantly (p>0.05) affected by the
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343
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344 mixing technique and gelatine dose.

345 As the control sample, honey jelly candy produced by cold mixing technique and

346 gelatine dose of 15% was decided. Cold mixing technique was chosen because diastase

347 number was significantly affected from mixing technique while it was insignificant in case of

348 sensorial quality. Additionally, minimum amount of gelatine dose was chosen to keep natural

349 properties and amount of honey at maximum level in the final product. Thus, the candy with
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350 15% of gelatine was re-produced by the cold mixing technique using fresh juices of orange,

351 strawberry and black mulberry instead of distilled water.

352

353 3.2. Fruity honey jelly candies

354 3.2.1. The effects of adding fruit juices on chemical properties of candies

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355 The physicochemical properties of fruity jelly candies are given in Table 1. The

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356 moisture content and water activity of samples were not significantly (p>0.05) affected by the

357 addition of fruit juices, and they changed between 20.05-25.68% and 0.70-0.75, respectively.

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358 However, fruity jelly candies descriptively had a lower water content and activity compared

359 to the control sample, which may be due to the additional colligative effect of the sugars in

360 the fruit juice.

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361 The titratable acidity and pH values of samples were significantly (p<0.05) affected by
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362 the addition of fruit juices. The highest pH and the lowest acidity value were determined in

363 control sample, produced without addition of fruit juice. The addition of fruit juice caused an
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364 increase in titratable acidity value of samples since they contain various organic acids. It was
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365 reported that the pH value of orange, strawberry and black mulberry were determined to be

366 3.35, 3.65 and 3.52, respectively (Grigelmo-Miguel and Martıń -Belloso, 1999; Ercisli and
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367 Orhan, 2007; Kelebek, Selli, Canbas, & Cabaroglu, 2009).

The diastase number of samples was not significantly (p>0.05) affected by the addition
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368
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369 of fruit juices and it was determined between 15.90-20.66. The diastase number of fruity jelly

370 candies was higher than the control due to the diastase enzymes arising from the fruit juices in

371 addition to honey.

372 The sugar profile and proline content of fruity jelly candies are shown in Table 2. While

373 the fructose and sucrose contents of samples were significantly (p<0.01) affected, the glucose

374 content of fruity jelly candies was not significantly (p>0.05) affected by the addition of fruit
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375 juices. The highest fructose content was determined in the strawberry fruity jelly candy as

376 593.05 g/kg and the highest sucrose content was determined in the orange fruity jelly candy as

377 40.86 g/kg. The sucrose content of strawberry and mulberry juice added candy samples (8.07

378 and 12.47 g/kg) were lower than orange juice added samples (40.86 g/kg). The differences in

379 terms of sugar content were due to the different fruit sources used. Some researchers reported

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380 comparatively different sugar contents of orange, strawberry and black mulberry (Eyduran et

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381 al., 2015; Kelebek, Selli, Canbas, & Cabaroglu, 2009; Mikulic‐Petkovsek et al., 2012) and it

382 was stated as the sugar content and composition of fruits can change depend on genetic and

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383 environmental factors, cultivation conditions, maturation degree and harvesting time

384 (Nikolaou et al., 2017). The glucose, fructose and sucrose contents of orange juice were

385
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reported as 32.30, 28.55 and 59.34 g/kg, respectively (Kelebek, Selli, Canbas, & Cabaroglu,
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386 2009). The glucose, fructose and sucrose contents of strawberry juice were 27.30, 30.20 and
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387 2.48 g/kg and black mulberry juice were 36.80, 39.90 g/kg and trace amount, respectively

388 (Mikulic‐Petkovsek et al., 2012). Additionally, fructose content of mulberry fruit was
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389 reported between 54.07 and 56.34 g/kg (Gundogdu et al., 2011), while this value could be
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390 reached to 90.97 g/kg in orange juice (Nikolaou et al., 2017).

391
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392 3.2.2. The effects of adding fruit juices on the physical properties of candies

The colour analysis results of the fruity jelly candies are presented in Table 3. The L*, a*
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393

and b* colour parameters were significantly (p<0.01) affected by the different fruit juices. It
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395 was observed that the L*and b* colour parameters were higher in the orange fruity jelly candy

396 and the a* colour parameter was higher in the strawberry and black mulberry fruity jelly

397 candies, which may be related to the different natural fruit juice colour.

398 The texture profile analysis results of the fruity jelly candies are shown in Table 4. The

399 texture parameters were not significantly (p>0.05) affected by the different fruit juices except
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400 for adhesiveness (p<0.01) and cohesiveness (p<0.05). The adhesiveness of candy with black

401 mulberry was higher, while its cohesiveness value was lower than the other samples. This

402 may be due to the high titratable acidity content of black mulberry fruity candy because of the

403 increment of surface polarity.

404 3.2.3. The effects of adding fruit juices on the sensorial properties of candies

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405 The sensory analysis results of the fruity jelly candies are presented in Figure 2.

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406 According to results, the fruity samples got more than 3.5 on 5-point hedonic scale and they

407 were evaluated as sensorial acceptable.

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408

409 4. Conclusion

410
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Nowadays, consumers prefer healthier foods, like honey, due to their bioactive
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411 components as they are becoming increasingly aware of healthy nutrition. Consequently, they
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412 demand, especially for their children, a decreased exposure to unhealthy foods such as

413 confectionery products. This study showed that a jelly candy which is healthier for children
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414 when compared to conventional jelly products in market since they are produced with
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415 synthetic flavouring and colouring agents, can be produced by minimal processing using

416 honey instead of sugars. The process temperature was suitable as 55°C both to protect
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417 diastase activity of honey and to ensure mixing gelatine at different doses (15-25%).

Furthermore, different fruit juices can be used to make this new honey jelly candy more
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418
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419 colourful, thereby more sensorial acceptable.

420

421 Acknowledgements

422 The authors would like to thank Akdeniz University and The Scientific and

423 Technological Research Council of Turkey (TUBITAK, project program: 2209-A) for

424 supporting this research.

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425

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1 Table 1. Physicochemical properties of honey jelly candies produced with different mixing
2 techniques, gelatine doses and fruit juices

Moisture Water Acidity Diastase

Mixing technique a pH
(%) activity (meq/kg) number
Cold 23.31a ± 2.82 0.73a ± 0.03 5.03a ± 0.02 40.12b ± 1.00 15.43a ± 0.87
Hot 23.45a ± 2.27 0.73a ± 0.02 4.91b ± 0.02 46.09a ± 1.39 0.00b ± 0.00

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Sign. - - ** ** **

Gelatine dose (%) b

15 23.32a ± 1.82 0.73a ± 0.02 4.91c ± 0.04 45.46a ± 1.98 7.95a ± 4.70

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20 25.08a ± 4.07 0.75a ± 0.04 4.97b ± 0.04 43.52ba ± 2.45 7.61a ± 4.44
25 21.75a ± 3.32 0.72a ± 0.04 5.02a ± 0.03 40.34b ± 1.26 7.59a ± 4.43

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Sign. - - ** * -
Fruity c

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Control 25.68a ± 1.83 0.75a ± 0.02 4.98a ± 0.02 42.12b ± 1.14 15.90a ± 2.51
Orange 23.76a ± 3.04 0.74a ± 0.03 4.31b ± 0.18 70.79a ± 5.56 16.29a ± 5.95
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Strawberry 20.05a ± 2.55 0.70a ± 0.04 4.43ba ±0.13 61.35a ± 5.42 20.66a ± 0.00
Black mulberry 20.34a ± 2.52 0.70a ± 0.03 4.26b ± 0.24 75.73a ± 0.46 19.53a ± 0.00
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3 Superscript letters beside the mean values denote in the same column that are significantly different by Duncan’s
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4 multiple range test. ** and * represent significance level at p ⩽ 0.01 and 0.01< p ⩽ 0.05, respectively.
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5 a; n = 6, b; n = 4, c; n = 2

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15 Table 2. Sugar and proline contents of honey jelly candies produced with different mixing

16 techniques, gelatine doses and fruit juices

Glucose Fructose Sucrose Proline

Mixing technique a
(g/kg) (g/kg) (g/kg) (mg/kg)
Cold 403.23a ± 5.24 491.10a ± 7.61 1.38a ± 0.12 110.27a ± 11.61
368.28b ± 4.53 454.70b ± 5.73 0.74b ± 0.09 74.21b ± 15.27

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Hot
Sign. ** ** ** **
Gelatine dose (%) b

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15 396.52a ± 12.96 488.40a ± 13.38 1.32a ± 0.25 130.28a ± 8.89
387.73b ± 7.74 475.14b ± 8.90 1.04b ± 0.10 86.85b ± 12.39

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25 373.02c ± 10.16 455.15c ± 10.64 0.83c ± 0.21 59.60c ± 12.86
Sign. ** ** ** **
Fruity c
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Control 418.79a ± 1.10 511.30c ± 5.12 1.41c ± 0.12 140.97b ± 15.54
Orange 419.59a ± 12.09 544.58b ± 9.87 40.86a ± 2.26 220.35ba ± 61.83
396.91a ± 0.71 593.05a ± 0.00 8.07b ± 0.73 304.61a ± 13.15
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Strawberry
Black mulberry 419.05a ± 0.00 439.38d ± 0.00 12.47b ± 0.34 222.47ba ± 5.81
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17 Superscript letters beside the mean values denote in the same column that are significantly different by Duncan’s
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18 multiple range test. ** and * represent significance level at p ⩽ 0.01 and 0.01< p ⩽ 0.05, respectively.

19 a; n = 6, b; n = 4, c; n = 2
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31 Table 3. Colour parameters of honey jelly candies produced with different mixing techniques,

32 gelatine doses and fruit juices

Mixing technique a L* a* b*
Cold 61.72a ± 1.21 8.70b ± 1.14 54.99a ± 1.62
Hot 48.63b ± 1.72 16.24a ± 1.42 45.06b ± 3.25

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Gelatine dose (%) b
57.31a ± 3.60 11.19a ± 2.09 51.85a ± 2.73

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20 55.14a ± 4.93 12.03a ± 2.03 51.59a ± 5.46
53.07a ± 3.32 14.19a ± 3.52 46.63a ± 3.80

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Sign. - - -
Fruity c
Control 63.08a ± 1.80
U 8.21b ± 2.91 55.37a ± 2.22
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Orange 64.24a ± 2.38 5.12b ± 0.69 56.75a ± 5.33
Strawberry 42.16b ± 1.33 25.86a ± 0.71 35.38b ± 1.39
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Black mulberry 28.77c ± 0.33 23.08a ± 3.00 9.95c ±1.93

Sign. ** ** **
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34 multiple range test. ** and * represent significance level at p ⩽ 0.01 and 0.01< p ⩽ 0.05, respectively.

35 a; n = 6, b; n = 4, c; n = 2
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Table 4. Texture parameters of honey jelly candies produced with different mixing techniques, gelatine doses and fruit juices

Mixing technique a Hardness (N) Adhesiveness (N.sec) Cohesiveness Springiness Chewiness Gumminess

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Cold 13.26a ± 1.69 0.08a ± 0.01 0.93a ± 0.01 0.96b ± 0.03 1199a ± 157 1251a ± 156
14.87a ± 1.11 0.03b ± 0.00 0.94a ± 0.01 1.05a ± 0.03 1485a ± 91 1425 a ± 102

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Hot
Sign. - ** - * - -

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Gelatine dose (%) b
15 10.47c ± 1.58 0.04b ± 0.01 0.93a ± 0.01 1.07a ± 0.04 1063b ± 192 1009b ± 155

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20 14.42b ± 0.55 0.05b ± 0.01 0.93a ± 0.01 1.01ba ± 0.02 1371ba ± 129 1401a ± 72

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25 17.30a ± 0.46 0.08a ± 0.03 0.94a ± 0.01 0.93b ± 0.04 1593a ± 42 1604a ± 62

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Fruity c

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Control 8.07a ± 0.39 0.06cb ± 0.02 0.92ba ± 0.01 1.00a ± 0.00 754.40a ± 27.61 769.53a ± 25

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Orange 9.22a ± 0.60 0.10b ± 0.01 0.94a ± 0.00 1.01a ± 0.03 809.27a ± 115.37 830.69a ± 111
Strawberry 7.94a ± 1.38 0.05c ± 0.01 0.94a ± 0.01 0.97a ± 0.01 959.82a ± 60.88 881.54a ± 51
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Black mulberry 7.15a ± 0.32 0.16a ± 0.01 0.90b ± 0.01 1.00a ± 0.02 951.85a ± 11.83 996.92a ± 133
Sign. - ** * - - -
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a; n = 6, b; n = 4, c; n = 2
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Sensorial score

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2 Figure 1. Sensory analysis results of honey jelly candies (Legend: Cold 15%; Cold 20%;
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3 Cold 25%; Hot 15%; Hot 20%; Hot 25%. Cold and Hot represent mixing

4 techniques and percentages represent gelatine doses)

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15 Figure 2. Sensory analysis results of control and fruity jelly candies produced with cold mixing
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16 technique including 15% gelatine dose (Legend: Control; Orange; Strawberry;

17 Black mulberry)
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Two mixing methods (cold and hot), three gelatine rates (15, 20, 25%) were studied

Cold mixing technique protected diastase enzyme activity more than 95%

Sugar and proline contents of cold mixed samples were higher than hot mixed samples

Cold mixed sample with 15% gelatine rate was selected as the best candy

Selected candy was reproduced by fresh orange, strawberry and black mulberry juices

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