Accepted Manuscript

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Detection of Quercetin Based on Al -Amplified Phosphorescence Signals of
Manganese-doped ZnS Quantum Dots

Zhifeng Zhang, Yanming Miao, Linwang Lian, Guiqin Yan

PII: S0003-2697(15)00378-4
DOI: 10.1016/j.ab.2015.08.002
Reference: YABIO 12156

To appear in: Analytical Biochemistry

Received Date: 10 April 2015

Revised Date: 26 July 2015
Accepted Date: 5 August 2015

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Please cite this article as: Z. Zhang, Y. Miao, L. Lian, G. Yan, Detection of Quercetin Based on Al -
Amplified Phosphorescence Signals of Manganese-doped ZnS Quantum Dots, Analytical Biochemistry
(2015), doi: 10.1016/j.ab.2015.08.002.

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 ACCEPTED MANUSCRIPT

1 Detection of Quercetin Based on Al3+-Amplified Phosphorescence

2 Signals of Manganese-doped ZnS Quantum Dots

3 Zhifeng Zhang , Yanming Miao, Linwang Lian, Guiqin Yan∗

4 Shanxi Normal University, Linfen, Shanxi 041000,China

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6 Short title: “Detection of Quercetin Based on Mn-doped ZnS QDs”

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8 Category: sensors
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Corresponding author. Fax: (86)0357-2051009. E-mail: gqyan2013@163.com.

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22 Abstract: A simple phosphorescence method is proposed for quercetin detection

23 based on Al3+-amplified Room-Temperature Phosphorescenee (RTP) signals of MPA

24 (3-Mercaptopropionic Acid)-capped Mn-doped ZnS quantum dots (QDs). The sensor

25 was established based on some properties as following: Al3+ can interact with

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26 carboxyl groups on the surface of MPA-capped Mn-doped ZnS QDs via chelation,

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27 which will lead to the aggregation of QDs and amplification of RTP signals, After the

28 addition of quercetin, it can form more stable complex with Al3+ in alkaline aqueous

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29 solution and dissociate Al3+ from the surface of Mn-doped ZnS QDs, which will result

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30 in significant recovery of RTP intensity of the MPA-capped Mn-doped ZnS-Al3+
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31 system. Under the optimized conditions, the change of RTP intensity was proportional

32 to the concentration of quercetin in the range from 0.1 to 6.0 mg L-1 with a high
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33 correlation coefficient of 0.996 and a detection limit of 0.047 mg L-1. The proposed
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34 method is potentially suitable for detection of quercetin in real samples without

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35 complicated pretreatment.

36 Keywords: Quantum dots, Signal amplification, Quercetin, detection.

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44 1. Introduction

45 As a new type of nanomaterial, Quantum dots (QDs) have attracted a lot of

46 attention in recent years owing to their unique properties, such as high quantum yield,

47 pronounced photostability and broad absorption spectra coupled with narrow and

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48 symmetrical emission spectra [1-3]. QDs are excellent optical material, which have

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49 been widely used to detect specific analytes, including ions, toxic molecules, small

50 molecules and biomacromolecules [4-6]. However, the construction of many

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51 analytical methods is based on the fluorescence properties of QDs. At present, more

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52 and more attention has been paid to the phosphorescence properties of long-lived
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53 room-temperature phosphorescence QDs [7-10]. Phosphorescence originates from the

54 triple state and has longer average life than fluorescence. Therefore, an appropriate
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55 delay time is allowed by phosphorescence and the interference from scattered light
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56 and autofluorescence can be effectively avoided accordingly [11]. Besides, the

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57 selectivity is also enhanced as it is less common than fluorescence [12, 13]. The RTP

58 QDs are endowed with potential superiority and applicability in complex chemical
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59 and biological analyses due to all of these properties.

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60 More interestingly, the large surface of QDs is favorable for attaching variable
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61 ligands. After the introduction of large-surface QDs, some ions, small molecules and

62 biomacromolecules will undergo physical or chemical reaction with QDs and further

63 change the structure or charge composition on the surfaces of the QDs, so some more

64 excellent controllable properties will be obtained accordingly [14]. Several studies

65 related to interactions between QDs and metal ions reveal that the surface capping

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66 ligands will profoundly affect the luminescence response of QDs to some metal

67 cations [15-19]. Detection methods established under the interaction between metal

68 ions and QDs are widely reported, but the sensing mechanisms of these methods are

69 mostly based on the fact that metal ions can quench the fluorescence or

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70 phosphorescence signals of QDs [20, 21]. Probes based on the quenching of

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71 fluorescence signals often suffer from very large background interference, which

72 results in a high detection limit. In comparison, probes based on emission

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73 enhancement only encounter very low background interference. Therefore, the

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74 development of probes based on enhancement of QD phosphorescence or
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75 fluorescence is very significant for improving the capability of detection.

76 Quercetin (3,3′,4′,5,7-penta hydroxyl flavone)(Fig.1) is one of the most abundant

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77 natural flavonoids, which is widely distributed in vegetables and fruits especially in

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78 traditional Chinese herbs and can reach in the human diet with a level of 16-25
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79 mg/day [22]. Over the past years, quercetin has not only drawn much attention

80 because of its various beneficial influences on human health, including anti-cancer,

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81 anti-inflammatory, anti-tumor, anti-ulcer, anti-allergy, anti-viral and anti-oxidant

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82 effects [23, 24] but also enable to protect human DNA from oxidative attack in
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83 vitro[25]. In spite of a high dietary intake of quercetin, only very low amounts was

84 excreted in the serum and urine of humans. For instance After the intake of quercetin

85 in human body, only 0.4-1% of quercetin can be excreted in urine [26-27]. And the

86 content of quercetin in serum is about 0.12-0.35mg L-1 after taken quercetin drug 500

87 mg [28]. Thus, a very sensitive method is required for determining quercetin in

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88 pharmaceutical drugs and biological samples.

89 At present, many highly sensitive and effective methods have been used to

90 determine the content of quercetin, such as liquid chromatography[29], liquid

91 chromatography [30, 31], spectrophotometry [32], mass spectrometry (LC-MS/MS)

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92 [33], Raman spectroscopy method [34], ionic liquids-based monolithic cartridge,

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93 molecular imprinting technique [35], electrochemical method[36-38] and so on.

94 However, some of these methods are limited by large time-consumption, technical

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95 complexity. For example, high performance liquid chromatography (HPLC) and

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96 HPLC-MS require complicated pretreatment. Despite high sensitivity and
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97 effectiveness, the electrochemical methods often require sophisticated electrode

98 modification. Hence, the development of simple, economical and sensitive analytical

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99 detection method has very higher application value.

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100 As a Lewis base, quercetin structurally possesses super delocalizability, a

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101 complete π bond conjugated system, strong coordination oxygen atoms, and

102 appropriate space configuration. Therefore, quercetin can selectively interact with
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103 Lewis acide Al3+ to form a stable chelate through acid-base adduct [39, 40]. Based on
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104 such interaction, quercetin was used to selectively determine and trace Al3+ [41-43],
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105 which was used as an effective probe to detect quercetin [44 45]. Moreover, it was

106 found that Al3+ can efficiently amplify the RTP signals of MPA-capped Mn-doped

107 ZnS QDs. Therefore, the ternary interaction among Al3+, MPA-capped Mn-doped ZnS

108 QDs and quercetin seems to be adopted as a basis for the development of RTP method

109 for quercetin detection.

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110 Inspired by these capabilities, we presented a simple quercetin detection system

111 based on the RTP of MPA-capped Mn-doped ZnS QDs. This sensing system was

112 composed of MPA-capped Mn-doped ZnS QDs and Al3+, without any sophisticated

113 process of functionalization or conjugation. Herein, Al3+ not only acted as

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114 coordination ions to amplify the RTP signals of MPA-capped Mn-doped ZnS QDs, but

also served as a probe to recognize quercetin. As illustrated in Fig. 1a, Al3+ can

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116 interact with MPA-capped Mn-doped ZnS QDs via covalent complexation and further

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117 lead to the aggregation of QDs and amplification of RTP intensity. Upon its addition,

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118 quercetin can compete with MPA-capped Mn-doped ZnS QDs to bind with Al3+ and
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119 form a more stable complex, and further detach Al3+ from the surface of MPA-capped

120 Mn-doped ZnS QDs. As a result, the RTP intensity of MPA-capped Mn-doped ZnS
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121 QDs will be recovered with the increase of quercetin concentration. Based on this
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122 strategy, the sensing system will be endowed with high selectivity and more
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123 convenience for quercetin detection. What’s important, this detection method provides

124 a basis of optical detection for other flavonoids since quercetin is a major
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125 representative of other flavonoids.

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126 Insert Figure 1 here

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127 2. Experiment section

128 2.1. Materials and apparatus

129 Mercaptopropionic acid (J&K Scientific, Beijing, China), Zn(Ac)2·2H2O,

130 Mn(Ac)2·4H2O, Na2S·9H2O, Al(NO3)3·9H2O were purchased from Tianjing Kermel

131 Chemical Reagent Co. (Tianjing, China). Quercetin was bought from Sigma-Aldrich

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132 Corporation. All other chemicals were of analytical grade and the resistivity of water

133 used in this study was higher than 18 M Ω·cm.

134 The morphology and microstructure of QDs were characterized by a JEM-2100F

135 transmission electron microscope (TEM, HRTEM, Japan) and a D8 Advanced X-ray

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136 diffractometer (Bruker, Germany, Cu Kα) respectively. Besides, samples for HRTEM

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137 were obtained by drying sample droplets from water dispersion onto a 300-mesh Cu

138 grid coated with a lacey carbon film. Fourier transform infrared (FT-IR) spectra

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139 (4000-400 cm-1) in KBr were recorded on a Magna-560 spectrometer (Nicolet,

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140 Madison, WI). Al(III)-induced aggregation of QDs was characterized by a JSM-7500F
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141 scanning electron microscope (SEM, JEOL, Japan). Moreover, Cary Eclipse

142 fluorescence spectrophotometer (Varian American Pty Ltd., USA) equipped with a
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143 plotter unit and quartz cell (1cm×1cm) in phosphorescence mode was used to record
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144 phosphorescence. The slit width was 10 nm and 20 nm for excitation (295 nm) and
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145 emission (590 nm) separately, and the scanning wavelength was in the range from 200

146 to 700 nm. In additional, resonance light scattering (RLS) spectra were recorded on
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147 the same spectrofluorometer by simultaneously scanning of excitation and emission

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148 monochromators (∆λ=2.5) from 200 to 700 nm, while dynamic light scattering (DLS)
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149 measurements were conducted on a Malvern Zetasizer Nano ZS90DLS system

150 (Malvern Instru-ments Ltd., Worcestershire, UK). The UV spectra were measured by

151 using a Shimadzu UV-29100 ultraviolet/visible (UV/Vis) spectrophotometer, and pH

152 was tested by a PHS-3C pH meter (Jinpeng Analytical Instruments Co. Ltd, China).

153 2.2. Synthesis of the Mn-Doped ZnS QDs

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154 MPA-capped Mn-Doped ZnS QDs were synthesized based on a published method

155 [11, 46] with minor modification. Please find the supplementary content from

156 supplementary material for detailed synthetic process.

157 2.3. Analytical procedures

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158 To study the amplifying effect of Al3+ on the RTP of MPA-capped Mn-doped ZnS

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159 QDs, first, we dissolved Al(NO3)3·9H2O in water to get a concentration of 10 mM.

160 Second, a series of samples with different concentrations were prepared by adding

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161 different amounts of Al3+ to Tris-HCl solution (pH 8.0, 20 mM). Third, the

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162 MPA-capped Mn-doped ZnS QDs were dissolved in water to get a concentration of 2
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163 mg mL-1 and then 100 µL of QD solution was added to each of above Al3+ solutions.

164 Finally, the mixtures were gently diluted to be 5 mL with ultrapure water and shaken
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165 for 15 min. at room temperature before detection.

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166 For quercetin determination, 100 µL of MPA-capped Mn-doped ZnS QDs (2 mg

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167 mL-1) and 70µL of Al3+ (10 mM) were prepared in Tris-HCl (20 mM, pH 8.0), then a

168 series of quercetin samples with different concentrations were added into each of
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169 above solutions after 15 min. The mixtures were gently diluted to be 5 mL with
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170 ultrapure water and shaken for 5 min. before analysis.

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171 2.4. Preparation of test real samples

172 The urine and serum samples used for preparing standard curves in practical

173 sample analysis were collected from healthy volunteer and diluted for 50-fold before

174 analysis. The serum and urine samples used for recovery experiment were collected

175 from volunteers 2 h later after they took 200 mg quercetin drug by oral medication

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176 and diluted for 50-fold before analysis.

177 3.Results and discussion

178 3. 1. Characterization of the MPA-capped Mn-doped ZnS QDs

179 The morphologies of MPA-capped Mn-doped ZnS QDs were characterized by

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180 TEM, X-ray diffraction (XRD) energy dispersive spectrum (EDX) analysis. Please

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181 find the characterization result from supplementary content.

182 3.2. Investigaton into amplifying effect of Al3+ on RTP of MPA-capped Mn-doped ZnS

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183 QDs

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184 The initial phosphorescence spectra of MPA-capped Mn-doped ZnS QDs were
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185 recorded with the absence or presence of Al3+. As shown in Fig. 2A, the MPA-capped

186 Mn-doped ZnS QDs had a weak RTP intensity, which can be increased by adding Al3+.
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187 Actually, the variation of RTP intensity was directly proportional to the concentration
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188 of Al3+ from 0 to 140 µM L-1 .When the concentration of Al3+ was above 140 µM L-1,
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189 the RTP intensity almost remained the same and became 2.7 times larger than that of

190 MPA-capped Mn-doped ZnS QDs at the same concentration (insert in Fig.2A). All of
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191 these results can prove that Al3+ is able to amplify the RTP signals of MPA-capped
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192 Mn-doped ZnS QDs.

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193 The MPA that caps Mn-doped ZnS QDs can not only enhance the water-solubility

194 of QDs, but also endow the surface of QDs with abundant carboxyl. Thereby, the RTP

195 of MPA-capped Mn-doped ZnS QDs was amplified by Al3+, which was probably

196 attributed to the interaction between Al3+ and the carboxyl in the surface of QDs. This

197 assumption can be ascertained by UV-vis absorption and FT-IR spectra. According to

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198 the UV-Vis absorption results (Fig.2B) a strong absorption of MPA-capped Mn-doped

199 ZnS QDs can be observed in the UV area at 240-320 nm (curve 1), while the UV

200 absorption of Al3+ was very weak (curve 2). However, the UV absorption of

201 MPA-capped Mn-doped ZnS QDs was significantly enhanced (curve 4) after the

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202 addition of Al3+ into Mn-doped ZnS QDs-Al3+ solutions, but such enhancement was

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203 not induced by the superposition of two absorptions (curve 3). Such result indicates

204 that an interaction between Al3+ and MPA-capped Mn-doped ZnS QDs was occurred

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205 to modestly change the surface structure of MPA-capped Mn-doped ZnS QDs. FT-IR

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206 spectra of MPA-capped Mn-doped ZnS QDs (curve 1) and MPA-capped Mn-doped
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207 Zn QDs-Al3+ (curve 2) are showed in figure 2C. In FT-IR spectra, the strong peak at

208 1569 cm-1 and 1398 cm-1 corresponds to the signifying of -C=O and -C-OH stretching,
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209 while the peak at 3428 cm-1 actually correspond to the signifying of -OH stretching.
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210 By comparison of FT-IR patterns (Fig. 2c curve 1 and 2), the stretching vibration of
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211 -C=O shifted from 1569 cm-1 to 1572 cm-1and the peaks around 3428 cm-1 were

212 obviously reduced in size with the presence of the Al3+. Additionally, the absorption
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213 bands at 854 cm-1 can be clearly observed, which can reflect the formation of Al-O.
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214 Overall, the above results can verify the covalent complexation of Al3+ on the surface
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215 of QDs.

216 Moreover, it can be seen from the RLS spectra (Fig. 2D.) that RLS intensity of

217 MPA-capped Mn-doped ZnS QDs at 200-700 nm was weak, which can be

218 significantly enhanced with the increase of Al3+ concentration after mixing with Al3+

219 (with a maximum at 395 nm). These results signify that the formation of larger

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220 scattering particles was caused by interaction between MPA-capped Mn-doped ZnS

221 QDs and Al3+. According to the DLS analysis, we can further find that (Fig. 2E), the

222 average hydrodynamic diameter (DH) of MPA-capped Mn-doped ZnS QDs is 8.84 nm

223 (inset Fig. 2E) and the DH can be increased from 8.84 to 36.78 nm after the addition

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224 of Al3+, which validate that the effective aggregation of Mn-doped ZnS QDs was

happened in the presence of Al3+ and thus shortened the distance between QDs.

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226 Similar phenomena was also reported here [47].

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227 Insert Figure 2 here

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228 As is well-known, many defects/traps appeared on the surface of QDs were
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229 considered as efficient nonradiative recombination centers. During the nonradiative

230 relaxation, the excitation energy is dissipated as heat or vibration, rather than emitted
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231 as light. When Al3+ is added to the solutions of MPA-capped Mn-doped ZnS QDs, the
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232 surface nonradiative relaxation centers are passivated efficiently by binding with Al3+.
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233 The passivation of surface trap sites are either ‘‘filled’’ or energetically approach to

234 the band edges, which effectively eliminate the nonradiative recombination of
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235 defects/traps [48]. As a result, a photoluminescence-activation effect that enhanced

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236 the QD photoluminescence is induced accordingly. What’s more, the following

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237 consequences might be induced by the reduction of between-QD distance [47, 49-52]:

238 (1) the defect of single QD can be repaired by the neighboring QDs; (2) the Mn ions

239 distributed on the surface of QDs can be rearranged to inner location, and thus result

240 in higher Mn2+ emission; (3) the between-QD Coulomb force will be enhanced, which

241 will improve the local electric field around the QDs and induce more effective

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242 excitation of QDs. Each aspect was attributed to the enhanced RTP emission of

243 MPA-capped Mn-doped ZnS QDs. All of these facts can explain why Al3+ amplifies

244 the RTP signals of MPA-capped Mn-doped ZnS QDs.

245 3.3.，Optimization of the Mn-doped ZnS QDs-Al3+ system

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246 To enhance the sensitivity and stability of proposed system, we investigated the

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247 effects of pH value of solution, reaction time and salt concentration on the

248 MPA-capped Mn-doped ZnS QDs-Al3+ system. As illustrated in Figure S2A, the～RTP

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249 intensity of MPA-capped Mn-doped ZnS QDs is increased gradually with the rising

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250 pH within 6.0-8.4, but it is relatively stable within a pH range from 7.0 to 8.0.
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251 Therefore, all the following experiments were carried out in the pH 8.0 of Tris-HCl

252 solution. As shown in figure S2B, the RTP intensity was almost balanced after 15 min.
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253 and remained stable for more than 40 min. Thus, the duration of 15 min. was adopted
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254 in the following experiments. The salt concentration within 0-0.03 M did not
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255 significantly influence the RTP intensity of MPA-capped Mn-doped ZnS QDs-Al3+

256 system (Fig. S2C).

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257 3.4. Quercetin-induced RTP quenching of MPA-capped Mn-doped ZnS QDs-Al3+

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258 system
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259 To validate the applicability of MPA-capped Mn-doped ZnS QDs-Al3+ system, we

260 investigated the influences of quercetin on RTP intensity of MPA-capped Mn-doped

261 ZnS QDs-Al3+ with the absence or presence of Al3+. It can be seen clearly from Figure

262 3A that the RTP intensity of MPA-capped Mn-doped ZnS QDs-Al3+ is decreased

263 gradually with the increasing concentration of quercetin. By comparison, the RTP

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264 intensity of MPA-capped Mn-doped ZnS QDs was not influenced by the quercetin

265 (Fig. S2) without addition of Al3+, which indicates that Al3+ plays a role of key bridge

266 between the MPA-capped Mn-doped ZnS QDs and quercetin.

267 As the high super delocalizability, a complete large π bonding conjugated system

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268 and strong coordination oxygen atoms were shown structurally, quercetin is very

likely to interact with Al3+ to develop stable quercetin-Al3+ complex. The UV/Vis

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270 spectra of free quercetin and Al3+-treated quercetin are presented in Fig. 3B. It can be

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271 seen that quercetin has two characteristic absorption bands within 240 280 nm

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272 (λmax=255) and 310 400 nm (λmax=357). The absorption bands within 240 280 nm
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273 corresponds to the cinnamoyl system of B ring and the other peak within 310 400

274 nm corresponds to the benzoyl system of A ring. Accompanied with significantly

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275 decrease in absorption, the absorption bands within 240 280 nm and 310 400 nm
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276 shifted to a longer wavelength after the addition of Al3+. The infrared spectra of free
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277 quercetin and Al3+-treated quercetin are presented in Fig. 3C and Fig. S4 respectively.

278 As shown in Fig. 3C the pure quercetin not only indicates an extremely broad band
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279 due to hydrogen bonded hydroxyl groups (υ –OH) appeared at 3455 cm-1, but also
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280 shows a characteristic carbonyl C=O absorption band at 1661 cm-1 (curve 1). The
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281 characteristic absorption bands appeared at 1528 cm-1 and 1452 cm-1 were attributed

282 to benzene ring bond stretching. The characteristic absorption at l388 cm-1 and

283 1276 cm-1 was assigned to υC3-OH and υ=C-O-C stretching vibration respectively.

284 However, the stretching vibration absorption peaks of A ring and B ring shifted from

285 1528 to 1494 cm-1 and from 1454 to 1426 cm-1 respectively after the addition of Al3+

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286 (curve 2). Meanwhile, the absorption peak of C=O moved from 1661 to 1640 cm-1,

287 while the absorption peak of υC3-OH migrated from 1388 to 1351 cm-1. In addition, the

288 two absorption peaks of ortho-dyhydroxyl transferred to 1322 and 1329 cm-1

289 separately and the absorption intensity of C-O in ortho-hydroxy (at 1168 cm-1) are

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290 obviously reduced. Furthermore, a medium-intensity absorption peak of Al-O

appeared at 645 cm-1 can be clearly observed. These results indicate that Al3+

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292 coordinate with 3-hydroxy-4-ketone group of B-ring and the 3′, 4′ ortho-dyhydroxyl

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293 of C-ring in quercetin and form stable composites accordingly.

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294 After changed the adding order of quercetin, quantum dots (QDs) and Al3+ (Fig.
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295 3D), we find that the RTP intensity of MPA-capped Mn-doped ZnS QDs is almost

296 unchanged if quercetin and QDs are added first and allowed to react for 5 min. before
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297 the addition of Al3+ (< 60 µM). However, the RTP intensity of MPA-capped
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298 Mn-doped ZnS QDs is improved along with the increase of aluminum ion
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299 concentration above the level of 60 µM. Based on these results, it can be seen that

300 quercetin can preferentially combine with Al3+ to form a stable complex at low
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301 concentration of Al3+, which further prevents the interaction between Al3+ and QDs.
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302 However, at high concentration of Al3+, the relatively excessive Al3+ will interact with
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303 QDs and enhance the RTP intensity of QDs consequently. As a result, we

304 preliminarily think that quercetin is more capable than QDs in binding with Al3+.

305 In order to further clarify the interaction mechanism between Al3+ and quercetin

306 in the MPA-capped Mn-doped ZnS QDs, we investigated the variation of particle

307 diameter of MPA-capped Mn-doped ZnS QDs -Al3+ system by adding quercetin. As

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308 illustrated in Fig. S5, the particle diameter of MPA-capped Mn-doped ZnS QDs-Al3+

309 system (from 36.78 to 24.69 nm) is apparently decreased by the introduction of

310 quercetin (5 mg L-1), which signifies that the ternary complex of MPA-capped

311 Mn-doped ZnS QDs-Al3+- quercetin was not formed. However, a more stable complex

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312 is established by quercetin and Al3+, which disintegrated Al3+ from the surface of

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313 MPA-capped Mn-doped ZnS QDs and result in depolymerization of QDs.

314 Insert Figure 3 here

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315 3.5 Detection of quercetin by the M PA-capped Mn-doped ZnS QDs-Al3+ system

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316 Furthermore, the linearity relation between quercetin concentration and
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317 photoluminescence (PL) intensity of MPA-capped Mn-doped ZnS QDs-Al3+ system

318 was further studied under the optimal conditions. The results show that the ∆RTP was
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319 almost linear with the quercetin concentration in the range of 0.1-6.0 mg L-1 (Fig. 4).
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320 The regression equation is ∆RTP=43.46 Cquercetin + 20.18 (regression coefficient =

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321 0.996), and the detection limit for quercetin is 0.047 mg L-1 (where s is the standard

322 deviation from11 continuous parallel detections of the standard deviation with 0.2 mg
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323 L-1 quercetin). For systems without quercetin or with 0.3 mg L-1 quercetin, those 11
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324 continuous parallel detections on phosphorescence intensity have a relative standard

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325 deviation of 4.6%.

326 Insert Figure 4 here

327 As shown in Table S1, the detection limit of this method is lower compared with

328 spectrophotometric method [32], Raman spectroscopy method [34], flow injection

329 chemiluminescence detection [53] and ionic liquids-based monolithic cartridge

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330 detection [54], but it is higher if compared with fluorescence method[44] and

331 electrochemical method[55-56]. In comparison with fluorescence approach and

332 electrochemical method, this system suffers from less background interference in

333 biological fluids and does not need sophisticated electrode modification.

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334 3.6. Selectivity, reproducibility and stability MPA-capped Mn-doped ZnS QDs-Al3+

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335 system

336 In order to evaluate the possible interferences in this system, we investigated the

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337 effects on the RTP intensity of MPA-capped Mn-doped ZnS QDs-Al3+ system with

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338 0.5 mg L-1 quercetin by adding some probably coexisting inorganic ions and organic
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339 compounds. The results summarized in Table S2 reflect that 500-fold of K+, 500-fold

340 of Na+, 150-fold of Ca2+, 500-fold Glucose, 150-fold of BSA, 300-fold of DNA,
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341 400-fold of L-Cys, 140-fold of L-Arg, 400-fold of L-Gly, and 70-fold of Catechin can
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342 hardly affected the detection of quercetin (Table S2). Thus, such method displays high
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343 selectivity for the quercetin detection of QDs-Al3+ system.

344 The different concentrations of quercetin (2.0 and 4.0mg L-1) in the linear range
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345 were analyzed in 5 independent series on the same day (intra-day ) and 5 consecutive
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346 days (inter-day) from measurements of each series (Table S3,S4). The low RSD
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347 values of intra-day and inter-day indicated that the developed method has a high

348 reproducibility. The stability of was also explored. After 7 days of storage, the

349 response to quercetin was tested each day with the response of the sensor decreasing

350 only 5% compared to the initial response, which confirms long-term stability.

351 3.7. Analysis of quercetin in Real samples

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352 In order to demonstrate the practical application of this method for quercetin

353 detection in biological media, 2% of human serum urine and samples in 20 mM

354 Tris-HCl buffer (pH 8.0) were used as the medium for further investigations. As

355 shown in Fig. 2C, the linear relationship between RTP intensity and quercetin

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356 concentrations were found to be ∆RTP=39.55 Cquercetin + 11.51 (R=0.996, the linear

range is 1.5-5.5 mg L-1) and ∆RTP=40.84 Cquercetin + 15.20 (R=0.996, the linear range

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357

358 is 1.5-5.5 mg L-1) in human serum and urine samples respectively. And the LOD is

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359 0.091 mg L-1 of quercetin in serum and 0.073 mg L-1 in urine. In addition, recovery

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360 experiments were carried out by adding 1 mg L-1 and 2 mg L-1 quercetin to 50-fold
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361 diluted serum and urine samples from volunteers who have taken 200mg of quercetin

362 drug. The results were listed in Table 1, from which we can see that the RSD was
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363 lower than 4.0% and the average recoveries of quercetin were in the range of
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364 95-106%. Therefore, it could be said that QDs-Al3+ system is suitable for quantitative
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365 detection of quercetin in human serum and urine samples after taken quercetin by oral

366 medication.
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367 Insert Table 1 here

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368 4. Conclusions
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369 In this study, a simple phosphorescence method was developed for the rapid

370 detection of quercetin. Under the optimum condition, the linear response to quercetin

371 was good in the range of 0.1- 6.0 mg mL-1 under the detection limit of 0.047 mg L-1,

372 although it is less sensitive if compared to other methods like fluorescence and

373 electrochemical detection. Moreover, the QDs-based RTP method does not need

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374 deoxidant or other inducers and sophisticated electrode modification, and it can

375 prevent interference from autofluorescence and the scattering light of matrix

376 encountered in spectrofluorometry. Therefore the present method is highly feasible

377 and can be readily used for rapid detection of quercetin in serum and urine after

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378 medication of quercetin.

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379 Acknowledgments

380 This work was supported by the Research Fund for the Doctoral Program of

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381 Higher Education of China (NO:20111404110002), Construction Program of

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382 Chemical Advantage and Key Discipline of Shanxi Province of China (NO:912019),
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383 The funds of Shanxi normal university (ZR1505) and School of Life

384 Science(SMYKZ-32)
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385
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396 References

397 [1] P. Zrazhevskiy, M. Sena, X. Gao, Designing multifunctional quantum dots for

398 bioimaging, detection, and drug delivery, Chem. Soc. Rev. 39(2010) 4326-54.

399 [2] A.M. Smith, S. Nie, Semiconductor nanocrystals: structure, properties, and band

PT
400 gap engineering, Accounts chem. res. 43(2010) 190-200.

RI
401 [3] H. Chen, L. Lin, H.F. Li, J.M. Lin, Quantum dots-enhanced chemiluminescence:

402 Mechanism and application, Coordin. Chem. Rev. (2014) 86-100.

SC
403 [4] H.P. Huang, J.J. Zhu, DNA aptamer-based QDs electrochemiluminescence

U
404 biosensor for the detection of thrombin, Biosens. bioelectron. 25(2009) 927-930.
AN
405 [5] Y. Song, Y. Li, Z.P. Liu, L.L. Liu, X.Y. Wang, X.G. Su, et al., A novel ultrasensitive

406 carboxymethyl chitosan-quantum dot-based fluorescence "turn on-off"

M

407 nanosensor for lysozyme detection, Biosens. bioelectron. 61(2014) 9-13.

D

408 [6] Y.Q. Hao, L. Liu, Y.F. Long, J.X. Wang, Y.N. Liu, F.M. Zhou, Sensitive
TE

409 photoluminescent detection of Cu2+ in real samples using CdS quantum dots in

410 combination with a Cu(2+)-reducing reaction, Biosens. bioelectron. 41(2013)

EP

411 723-729.
C

412 [7] H.F. Wang, Y.Y. Wu, X.P. Yan, Room-temperature phosphorescent discrimination
AC

413 of catechol from resorcinol and hydroquinone based on sodium tripolyphosphate

414 capped Mn-doped ZnS quantum dots, Anal. chem. 85(2013) 1920-1925.

415 [8] L. Tan, C. Huang, R.F. Peng, Y.W. Tang, W.M. Li, Development of hybrid

416 organic-inorganic surface imprinted Mn-doped ZnS QDs and their application as

417 a sensing material for target proteins, Biosens. bioelectron. 61(2014) 506-511.

19
 ACCEPTED MANUSCRIPT

418 [9] E. Sotelo-Gonzalez, M.T. Fernandez-Arguelles, J.M. Costa-Fernandez, A.

419 Sanz-Medel, Mn-doped ZnS quantum dots for the determination of acetone by

420 phosphorescence attenuation, Anal. chim. acta, 712(2012) 120-126.

421 [10] Y.Q. Wang, W.S. Zou, 3-Aminopropyltriethoxysilane-functionalized manganese

PT
422 doped ZnS quantum dots for room-temperature phosphorescence sensing

RI
423 ultratrace 2,4,6-trinitrotoluene in aqueous solution, Talanta, 85(2011) 469-475.

424 [11] P. Wu, Y. He, H.F. Wang, X.P. Yan, Conjugation of Glucose Oxidase onto

SC
425 Mn-Doped ZnS Quantum Dots for Phosphorescent Sensing of Glucose in

U
426 Biological Fluids, Anal. Chem. 82(2010) 1427-1433.
AN
427 [12] H.F. Wang, Y. He, T.R. Ji, X.P. Yan, Surface Molecular Imprinting on Mn-Doped

428 ZnS Quantum Dots for Room-Temperature Phosphorescence Optosensing of

M

429 Pentachlorophenol in Water, Anal. Chem. 81(2009) 1615-1621.

D

430 [13] J.M. Traviesa-Alvarez, I. Sanchez-Barragan, J.M. Costa-Fernandez, R. Pereiro, A.

TE

431 Sanz-Medel, Room temperature phosphorescence optosensing of benzo[a]pyrene

432 in water using halogenated molecularly imprinted polymers, Analyst, 132(2007)

EP

433 218-223.
C

434 [14] W.E. Mahmoud, Functionalized ME-capped CdSe quantum dots based
AC

435 luminescence probe for detection of Ba2+ ions, Sensor. Actuator. B-Chem.

436 164(2012) 76-81.

437 [15] Y.H. Chan, J. Chen, Q. Liu, S.E. Wark, D.H. Son, J.D. Batteas, Ultrasensitive

438 copper(II) detection using plasmon-enhanced and photo-brightened

439 luminescence of CdSe quantum dots, Anal. chem. 82(2010) 3671-3678.

20
 ACCEPTED MANUSCRIPT

440 [16] T.W. Sung, Y.L. Lo, Highly sensitive and selective sensor based on silica-coated

441 CdSe/ZnS nanoparticles for Cu2+ ion detection, Sensor. Actuator. B-Chem.

442 165(2012) 119-125.

443 [17] T.W. Sung, Y.L. Lo, I.L. Chang, Highly sensitive and selective fluorescence

PT
444 probe for Cr3+ ion detection using water-soluble CdSe QDs, Sensor. Actuator.

RI
445 B-Chem. 202(2014) 1349-1356.

446 [18] Y. Shen, S. Liu, J. Yang, L. Wang, X. Tan, Y. He, A novel and sensitive turn-on

SC
447 fluorescent biosensor for the DNA detection using Sm3+-modulated

U
448 glutathione-capped CdTe quantum dots, Sensor. Actuator. B-Chem. 199(2014)
AN
449 389-397.

450 [19] W.E. Mahmoud, S.J. Yaghmour, A.M. Al-Amri, Enhancement of CdSe quantum
M

451 dots luminescence by calcium ions, Journal of Luminescence, 134(2013)

D

452 429-431.
TE

453 [20] R.Z. Zhang, W. Chen, Nitrogen-doped carbon quantum dots: facile synthesis and

454 application as a "turn-off" fluorescent probe for detection of Hg2+ ions, Biosens.
EP

455 bioelectron. 55(2014) 83-90.

C

456 [21] K. Zhang, J.k. Guo, J.j. Nie, B.y. Du, D.j. Xu, Ultrasensitive and selective
AC

457 detection of Cu2+ in aqueous solution with fluorescence enhanced CdSe quantum

458 dots, Sensor. Actuator. B-Chem. 190(2014) 279-287.

459 [22] M.J. Ko, C.I. Cheigh, S.W. Cho, M.-S. Chung, Subcritical water extraction of

460 flavonol quercetin from onion skin, J. Food Eng. 102(2011) 327-333.

461 [23] A.P. Rogerio, C.L. Dora, E.L. Andrade, J.S. Chaves, L.F. Silva, E. Lemos-Senna,

21
 ACCEPTED MANUSCRIPT

462 et al., Anti-inflammatory effect of quercetin-loaded microemulsion in the airways

463 allergic inflammatory model in mice, Pharmacol. res. 61(2010) 288-397.

464 [24] F. Dajas, Life or death: neuroprotective and anticancer effects of quercetin, J.

465 ethnopharmacol. 143(2012) 383-396.

PT
466 [25] P. Xiao, F. Zhao, B. Zeng, Voltammetric determination of quercetin at a

RI
467 multi-walled carbon nanotubes paste electrode, Microchem. J. 85(2007) 244-249.

468 [26] K.Ishii, T.Furuta, Y. Kasuya. High-performance liquid chromatographic

SC
469 determination of quercetin in human plasma and urine utilizing solid-phase

U
470 extraction and ultraviolet detection. J. Chromatogr. B,794 (2003) 49-56.
AN
471 [27]P .C.H. Hollman, M.N. Bijsman, Y. van Gameren, E.P. Cnossen, J.H. de Vries,

472 M.J. B Katan, The sugar moiety is a major determinant of the absorption of
M

473 dietary flavonoid glycosides in man. Free Rad. Res. 31(1999) 569-573.
D

474 [28]L.Wang, M.E. Morris. Liquid chromatography–tandem mass spectroscopy assay

TE

475 for quercetin and conjugated quercetin metabolites in human plasma and urine. J.

476 Chromatogr. B, 821 (2005) 194-201.

EP

477 [29] B.C. Prasougsidh, G.R. Skurray, Capillary electrophoresis analysis of trans- and
C

478 cis-resveratrol, quercetin, catechin and gallic acid in wine, Food Chem.
AC

479 62(1998) 355-358.

480 [30] K. Ishii, T. Furuta, Y. Kasuya, High-performance liquid chromatographic

481 determination of quercetin in human plasma and urine utilizing solid-phase

482 extraction and ultraviolet detection, J. Chromatogr. B, 794(2003) 49-56.

483 [31] E. Ranjbari, P. Biparva, M.R. Hadjmohammadi, Utilization of inverted dispersive

22
 ACCEPTED MANUSCRIPT

484 liquid-liquid microextraction followed by HPLC-UV as a sensitive and efficient

485 method for the extraction and determination of quercetin in honey and biological

486 samples, Talanta, 89(2012) 117-123.

487 [32] N. Pejic, V. Kuntic, Z. Vujic, S. Micic, Direct spectrophotometric determination

PT
488 of quercetin in the presence of ascorbic acid, Il Farmaco, 59(2004) 21-24.

RI
489 [33] L. Wang, M.E. Morris, Liquid chromatography-tandem mass spectroscopy assay

490 for quercetin and conjugated quercetin metabolites in human plasma and urine, J.

SC
491 chromatogr. B, 821(2005) 194-201.

U
492 [34] Y. Numata, H. Tanaka, Quantitative analysis of quercetin using Raman
AN
493 spectroscopy, Food Chem. 126(2011) 751-755.

494 [35] J.M. Liu, L.P. Lin, X.X. Wang, W.L. Cai, L.H. Zhang, S.Q. Lin, A highly
M

495 sensitive coupling technique for the determination of trace quercetin based on
D

496 solid substrate room temperature phosphorimetry and poly (vinyl alcohol)
TE

497 complex imprinting, Anal. chim. acta 723(2012) 76-82.

498 [36] Z. Zhang, S.Q. Gu, Y.P. Ding, M.J. Shen, L. Jiang, Mild and novel
EP

499 electrochemical preparation of beta-cyclodextrin/graphene nanocomposite film

C

500 for super-sensitive sensing of quercetin, Biosens. bioelectron. 57(2014) 239-244.

AC

501 [37] Y.Y. Yao, L.H. Zhang, Z.F. Wang, J.K. Xu, Y. Wen, Electrochemical

502 determination of quercetin by self-assembled platinum nanoparticles/poly

503 (hydroxymethylated-3,4-ethylenedioxylthiophene) nanocomposite modified

504 glassy carbon electrode, Chinese Chem. Lett. 25(2014) 505-510.

505 [38] M. Veerapandian, Y.T. Seo, K. Yun, M.H. Lee, Graphene oxide functionalized

23
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506 with silver@silica-polyethylene glycol hybrid nanoparticles for direct

507 electrochemical detection of quercetin, Biosens. bioelectron. 58(2014) 200-204.

508 [39] J.P. Cornard, J.C. Merlin, Spectroscopic and structural study of complexes of

509 quercetin with Al(III), J. Inorg. Biochem. (2002) 19-27.

PT
510 [40] A. Ahmedova, K. Paradowska, I. Wawer, 1H, 13C MAS NMR and DFT GIAO

RI
511 study of quercetin and its complex with Al(III) in solid state, J.Inorg. Biochem,

512 110(2012) 27-35.

SC
513 [41] H.Z. Lian, Y.F. Kang, S.P. Bi, Y. Arkin, D.L. Shao, D.N. Li, et al., Direct

U
514 determination of trace aluminum with quercetin by reversed-phase high
AN
515 performance liquid chromatography, Talanta, 62(2004) 43-50.

516 [42] P. Norfun, T. Pojanakaroon, S. Liawraungrath, Reverse flow injection

M

517 spectrophotometric for determination of aluminium(III), Talanta, 82(2010)

D

518 202-207.
TE

519 [43] Y. Zou, F. Yan, yong, L. Dai, feng, Y. Luo, mei, Y. Fu, N. Yang, et al., High

520 photoluminescent carbon nanodots and quercetin-Al3+ construct a ratiometric

EP

521 fluorescent sensing system, Carbon, 77(2014) 1148-1156.

C

522 [44] S.Y. Hung, W.L. Tseng, A polyadenosine-coralyne complex as a novel

AC

523 fluorescent probe for the sensitive and selective detection of heparin in plasma,

524 Biosens. bioelectron. 57(2014) 186-191.

525 [45] H. Wu, J. Wu, C. Saez, M. Campana, E.G. Megehee, E. Wang, Luminescence

526 response of an osmium(II) complex to macromolecular polyanions for the

527 detection of heparin and chondroitin sulfate in biomedical preparations, Anal

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528 Chim Acta, 804(2013) 221-227.

529 [46] J. Zhuang, X. Zhang, G. Wang, D. Li, W. Yang, T. Li, Synthesis of water-soluble

530 ZnS : Mn2+ nanocrystals by using mercaptopropionic acid as stabilizer, J. Mater.

531 Chem. 13(2003) 1853-1857.

PT
532 [47] H. Yan, H.F. Wang, Turn-on room temperature phosphorescence assay of heparin

RI
533 with tunable sensitivity and detection window based on target-induced

534 self-assembly of polyethyleneimine capped Mn-doped ZnS quantum dots, Anal.

SC
535 chem. 83(2011) 8589-8595.

U
536 [48] Q. Ma, Z.H. Lin, N. Yang, Y. Li, X.G. Su, A novel carboxymethyl
AN
537 chitosan-quantum dot-based intracellular probe for Zn2+ ion sensing in prostate

538 cancer cells, Acta biomater. 10(2014) 868-874.

M

539 [49] O. Kulakovich, N. Strekal, A. Yaroshevich, S. Maskevich, S. Gaponenko, I.

D

540 Nabiev, et al., Enhanced Luminescence of CdSe Quantum Dots on Gold Colloids,
TE

541 Nano Lett. 2(2002) 1449-1452.

542 [50] P. Anger, P. Bharadwaj, L. Novotny, Enhancement and quenching of

EP

543 single-molecule fluorescence, Phys. rev. lett. 96(2006) 113002.

C

544 [51] P.H. Borse, D. Srinivas, R.F. Shinde, S.K. Date, W. Vogel, S.K. Kulkarni, Effect
AC

545 of Mn2+ concentration in ZnS nanoparticles on photoluminescence and

546 electron-spin-resonance spectra, Phys. Rev. B, (1999) 8659-8664.

547 [52] Y. Hou, J. Ye, Z. Gui, G. Zhang, Temperature-Modulated Photoluminescence of

548 Quantum Dots, Langmuir, 24(2008) 9682-9685.

549 [53] H.M. Qiu, C.N. Luo, M. Sun, F.G. Lu, L.L. Fan, X.J. Li, A novel

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550 chemiluminescence sensor for determination of quercetin based on molecularly

551 imprinted polymeric microspheres, Food Chem. 134(2012) 469-473.

552 [54] T. Zhu, W.T. Bi, K. Row, Extraction and Determination of Quercetin and

553 Myricetin from Chamaecyparis obtusa by Ionic Liquids-based Monolithic

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554 Cartridge, Chinese J. Chem. 29(2011) 1759-1763.

RI
555 [55] M.L. Yola, V.K. Gupta, T. Eren, A.E. Sen, N. Atar, A novel electro analytical

556 nanosensor based on graphene oxide/silver nanoparticles for simultaneous

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557 determination of quercetin and morin. Electrochim. Acta 120 (2014) 204-211.

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558 [56]J.Manokaran, R.Muruganantham, A.Muthukrishnaraj, N.Balasubramanian.
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559 Platinum- polydopamine @SiO2 nanocomposite modified electrode for the

560 electrochemical determination of quercetin. Electrochim. Acta 168 (2015) 16-24.

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572 Figure captions:

573 Fig.1. The schematic illustration of fabricating Mn-doped ZnS QDs for quercetin

574 detection.

575 Fig.2. (A) RTP emission spectra of MPA-capped Mn-doped ZnS QDs in the presence

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576 of different concentrations of Al3+: (a) 20, (b) 40, (c) 60, (d) 80, (e) 100, (f) 120, (g)

140, (h) 160, (i) 180 and (j) 200 µM. The inset displays the plot of ∆RTP against Al3+

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577

578 concentrations. (B) UV-vis absorption spectra of Al3+ (curve 1), MPA-capped

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579 Mn-doped ZnS QDs (curve 2), MPA-capped Mn-doped ZnS QDs+Al3+(curve 3) and

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580 MPA-capped Mn-doped ZnS QDs-Al3+ system (curve 4).The concentration of
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581 MPA-capped Mn-doped ZnS QDs and Al3+ are 40 mg L-1 and 140 µM, respectively.

582 (C) FT-IR spectra of MPA Mn-doped ZnS QDs (curve 1) and MPA Mn-doped ZnS
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583 QDs/Al3+ complexes (curve 2). (D) RLS spectra of the MPA-capped Mn-doped ZnS
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584 QDs (40mg L-1) in the presence of different concentrations of Al3+: (a) 0, (b) 40, (c)
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585 60, (d) 80, (e) 100, (f) 120, (g) 140 and (h) 160 µM. (E) the adding order of quercetin,

586 quantum dots (QDs) and Al3+. (E) Size distribution MPA-capped Mn-doped ZnS QDs
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587 (QDs, 40 mg L-1µM,) and in the presence of Al3+ (140 µM ).

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588 Fig.3. (A) The RTP emission of the MPA-capped Mn-doped ZnS QDs-Al3+ system in
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589 the presence of different concentrations of quercetin: (a) 0, (b) 0.1, (c) 0.5, (d) 1.0, (e)

590 2.0, (f) 3.0, (g) 4.0, (h) 5.0, (i) 6.0, (j) 7.0 and (k) 8.0 mg L-1. (B) UV-vis absorption

591 spectra of Al3+(curve1), quercetin (curve2), quercetin+Al3+(curve 3) and

592 quercetin/Al3+ complexes (curve 4).The concentration of MPA Mn-doped ZnS QDs

593 and Al3+ are 40 mg L-1 and 140 µM, respectively. (C) FT-IR spectra of quercetin and

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594 quercetin/Al3+ complexes. (D) Effect of the adding order on the RTP intensity of the

595 MPA-capped Mn-doped ZnS QDs.

596 Fig. 4. The relation ship between ∆RTP and the concentration of quercetin (from 0.1

597 to 6.0 mg L-1);

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598

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600

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616 Fig. 1.

Al3+
O
S S O
O
HO O
O O O
O O O HO S
S OH Al3+ OH O +
Quercetin Al3+ S
S
Al3+ HO
OH O OH

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O O O O
O O
O
O OH
S S O
O

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Al3+

Low RTP intensity RTP signals amplificating RTP signals recovery

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OH
3' OH
O
1 C 4'
S OH HO O
: MPA-capped Mn-doped ZnS QDs

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7 A B
S OH 3
OH
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O 5
OH O

617 Quercetin
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618 Fig. 1. Schematic illustrating the sensing mechanism of MPA-capped Mn-doped ZnS

619 QDs- Al3+ system for quercetin detection.

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630 Fig. 2.

(A) (B) QDs

j 600

△RTP
800 3+
400 0.6 Al
3+
RTP intensity

3+ 200 4 QDs+Al

Absorbance
[Al ] 3
600 QDs/Al
3+

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0
0 50 100 150 200 0.4 2
400 a C (µM)

0.2

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200
1
0 0.0

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500 550 600 650 700 750 800 240 280 320 360 400

631 Wavelength (nm) Wavelength ( nm )

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(C) (D)
1569 1398 h
600
Transmittance (a.u.)

curve1
3428
RLS intensity

3+
[Al ]
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400

curve 2 a
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200
854
3422
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1561 1389 0

4000 3000 2000 1000 200 300 400 500 600 700
-1
Wavenumber (cm ) Wavelength (nm)
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632
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(E) 40
50 diameter: 8.84 nm
30
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40 20
Frequency (%)

10
30 0
6 9 12 15

20 diameter: 36.78 nm

10

0
10 20 30 40 50 60 70

633 Size (d. nm)

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634 Fig.2. (A) RTP emission spectra of MPA-capped Mn-doped ZnS QDs in the presence

635 of different concentrations of Al3+: (a) 20, (b) 40, (c) 60, (d) 80, (e) 100, (f) 120, (g)

636 140, (h) 160, (i) 180 and (j) 200 µM. The inset displays the plot of ∆RTP against Al3+

637 concentrations. (B) UV-vis absorption spectra of Al3+ (curve 1), MPA-capped

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638 Mn-doped ZnS QDs (curve 2), MPA-capped Mn-doped ZnS QDs+Al3+(curve 3) and

MPA-capped Mn-doped ZnS QDs-Al3+ system (curve 4).The concentration of

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639

640 MPA-capped Mn-doped ZnS QDs and Al3+ are 40 mg L-1 and 140 µM, respectively.

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641 (C) FT-IR spectra of MPA Mn-doped ZnS QDs (curve 1) and MPA Mn-doped ZnS

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642 QDs/Al3+ complexes (curve 2). (D) RLS spectra of the MPA-capped Mn-doped ZnS
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643 QDs (40mg L-1) in the presence of different concentrations of Al3+: (a) 0, (b) 40, (c)

644 60, (d) 80, (e) 100, (f) 120, (g) 140 and (h) 160 µM. (E) the adding order of quercetin,
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645 quantum dots (QDs) and Al3+. (E) Size distribution MPA-capped Mn-doped ZnS QDs
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646 (QDs, 40 mg L-1µM,) and in the presence of Al3+ (140 µM ).

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656 Fig. 3.

(A) (B) 0.5

800 a Quercetin/Al3+
0.4 Quercetin
[Quercetin] Al3+
RTP intensity

600

Absotance
0.3

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K
400 0.2

0.1

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0.0
0

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657 Wavelength (nm) Wavelenglh (nm)

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3428 1661 1528 1454 1276
Transmittance (a.u.)

1.5
RTP intensity (a.u.)

curve1
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1.0 600
RTP intensity

1388
400
0.5
D

200
curve2 1351
1492 0
0.0
500 600 700
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3435 1640 645

Wavelength (nm)

4000 3000 2000 1000 0 20 40 60 80 100 120 140

-1
Wavenumber (cm ) Concentration(µM)
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658

659 Fig.3. (A) The RTP emission of the MPA-capped Mn-doped ZnS QDs-Al3+ system in
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660 the presence of different concentrations of quercetin: (a) 0, (b) 0.1, (c) 0.5, (d) 1.0, (e)
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661 2.0, (f) 3.0, (g) 4.0, (h) 5.0, (i) 6.0, (j) 7.0 and (k) 8.0 mg L-1. (B) UV-vis absorption

662 spectra of Al3+(curve1), quercetin (curve2), quercetin+Al3+(curve3) and quercetin/Al3+

663 complexes (curve 4).The concentration of MPA Mn-doped ZnS QDs and Al3+ are 40

664 mg L-1 and 140 µM, respectively. (C) FT-IR spectra of quercetin and quercetin/Al3+

665 complexes. (D) Effect of the adding order on the RTP intensity of the MPA-capped

666 Mn-doped ZnS QDs.

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667 Fig. 4.

350
300
250

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200
150

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100
50

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0 2 4 6 8 10

668 Concentration (mg L-1)

669
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Fig. 4. The relation ship between ∆RTP and the concentration of quercetin (from 0.1
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670 to 6.0 mg L-1);
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683 Table captions:

684 Table 1

685 Results of quercetin detection in real samples

686

PT
687

RI
688

689

SC
690

U
691
AN
692

693
M

694
D

695
TE

696

697
EP

698
C

699
AC

700

701

702

703

704

34
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705 Table 1

706 Recovery for the detection of quercetin in real samples (Mean ± SEM; n=3)

Type of Found Added Total found Recovery RSD

samples (mg L-1) (mg L-1) (mg L-1) (%) (%, n=3)
1.0 1.14 98 3.9
Human urine 0.165
2.0 2.23 103 2.7

PT
1.0 1.16 96 4.4
Human serum 0.203
2.0 2.34 106 3.6

RI
707

SC
708

709

U
710
AN
711
M

712

713
D

714
TE

715
EP

716

717
C

718
AC

719

720

721

722

723

35
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724 Supplementary information

725

726 Supplementary content

727 Synthesis of the Mn-Doped ZnS QDs

PT
728 MPA-capped Mn-Doped ZnS QDs were synthesized based on a published method

RI
729 [11, 43] with minor modification. In brief, 10 mL of 0.1 M Zn(Ac)2, 4 mL of 0.01 M

730 Mn(Ac)2, and 100 mL of 0.04 M MPA were added to a three-neck flask. The solution

SC
731 was mixed and the pH value was adjusted to be 11 with 1 M NaOH under N2

U
732 atmosphere, then 10mL of 0.1 M Na2S was immediately injected into the solution
AN
733 after the air was removed with N2 bubbling for 30 min. Then the solution was aged at

734 50 °C in open air for 2 hours to form MPA-capped Mn-doped ZnS QDs after 20 min.
M

735 The QDs were purified by precipitation with ethanol, separated by centrifugation,
D

736 washed with ethanol and dried in vacuum. Such kind of as-prepared QD powder is
TE

737 highly soluble in water.

738 Characterization of the MPA-capped Mn-doped ZnS QDs

EP

739 The morphologies of MPA-capped Mn-doped ZnS QDs were characterized by

C

740 TEM. Clearly, these nanoparticles are nearly spherical and uniform with an average
AC

741 diameter of 3.5 nm (Fig. S1A). Meanwhile, the X-ray diffraction (XRD) spectra was

742 displayed, and a zinc-blend structure was typically exhibited by the prepared QDs as

743 revealed by the distinguishable (111), (220) and (311) planes in XRD patterns (Fig.

744 S1B). In addition, the energy dispersive spectrum (EDX) analysis showed that the

745 MPA-capped Mn-doped ZnS QDs are composed of Zn, S and Mn (Fig. S1C), while

36
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746 the phosphorescence spectra of QDs sensor indicate a maximum excitation peak at

747 295 nm and a narrow emission band around 590 nm (Fig. S1D), which is produced by

748 the energy transferred from the band gap ZnS to the dopant Mn2+ as well as the

749 subsequent transition from triplet state (4T1) to ground state (6A1) of Mn2+

PT
750 incorporated into the lattice of host ZnS[43]. All these results reveal that MPA-capped

RI
751 Mn-doped ZnS QDs were prepared successfully.

752

SC
753

U
754
AN
755

756
M

757
D

758
TE

759

760
EP

761
C

762
AC

763

764

765

766

767

37
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768 Supplementary Figure captions:

769 Fig. S1.(A)TEM image of MPA-capped Mn-doped ZnS QDs. (B) XRD image of

770 MPA-capped Mn-doped ZnS QDs. (C) EDS spectrum of MPA-capped Mn-doped ZnS

771 QDs. (dD) The excitation (curve a) and RTP emission (curve b) spectra of

PT
772 MPA-capped Mn-doped ZnS QDs ( 40 mg L-1) in Tris-HCl buffer (20 mM, pH 8.0).

Fig. S2. (A) Effect of pH on the RTP intensity of 40 mg L-1 MPA-capped Mn-doped

RI
773

774 ZnS QDs after addition of 140 µM Al3+. (B) Effect of reaction time on RTP intensity

SC
775 of 40 mg L-1 MPA-capped Mn-doped ZnS QDs after addition of 140 µM Al3+. (C)

U
776 Effect of NaCl concentration on the RTP intensity of 40 mg L-1 MPA-capped
AN
777 Mn-doped ZnS QDs after addition of 140 µM Al3+.

778 Fig. S3. Effect of quercetin concentration (0.05- 6 mg L-1) on the RTP intensity of the
M

779 MPA-capped Mn-doped ZnS QDs.

D

780 Fig. S4. FT-IR spectra of quercetin and quercetin/Al3+ complexes.

TE

781 Fig. S5. Size distribution MPA-capped Mn-doped ZnS QDs (40 mg L-1µM) and in the

782 presence of Al3+ (140 µM ).

EP

783 Fig. S6. The relation ship between ∆RTP and the concentration of quercetin (from
C

784 0.15 to 5.5 mg L-1) in Real samples (A: serum; B: urine).

AC

785

786

787

788

789

38
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790 Fig. S1.

A
(B)2000

1500

Intensity

PT
1000

500

RI
0
10 20 30 40 50 60 70 80

SC
791 2 Theta [deg.]

U
(D) 350
(C) 2000
AN
Conduction band
300
1600 tion
defect 4
250 T1
Excita
Intensity (a.u.)

Mn2+ hν2' RTP

Intensity

1200 200 hν1 6

A1
M

Valence band
S 150
800 Zn
Zn 100
b
D

400 a
Mn Mn Zn
50

0 0
TE

0 5 10 15 20 200 300 400 500 600 700

792 Energy (kev) Wavelength (nm)

EP

793 Fig. S1. (A)TEM image of MPA-capped Mn-doped ZnS QDs. (B) XRD image of

794 MPA-capped Mn-doped ZnS QDs. (C) EDS spectrum of MPA-capped Mn-doped ZnS
C
AC

795 QDs. (dD) The excitation (curve a) and RTP emission (curve b) spectra of

796 MPA-capped Mn-doped ZnS QDs ( 40 mg L-1) in Tris-HCl buffer (20 mM, pH 8.0).

797

798

799

800

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 ACCEPTED MANUSCRIPT

801 Fig. S2.

(A) (B) 1.2

1000
1.0
RTP intensity

800

PT
0.8

F/F0
600
0.6
400

RI
0.4
200

SC
5.5 6.0 6.5 7.0 7.5 8.0 8.5 0 10 20 30 40 50 60

802 pH value Time (min)

U
AN
(C) 1.2

1.0
M

0.8
F/F0

0.6
D

0.4
TE

0.2
0.0 0.5 1.0 1.5

Concentration (mM)
EP

803

804 Fig. S2. (A) Effect of pH on the RTP intensity of 40 mg L-1 MPA-capped Mn-doped
C

805 ZnS QDs after addition of 140 µM Al3+. (B) Effect of reaction time on RTP intensity
AC

806 of 40 mg L-1 MPA-capped Mn-doped ZnS QDs after addition of 140 µM Al3+. (C)

807 Effect of NaCl concentration on the RTP intensity of 40 mg L-1 MPA-capped

808 Mn-doped ZnS QDs after addition of Al3+ (140 µM).

809

810

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 ACCEPTED MANUSCRIPT

811 Fig. S3.

350
300
RTP intensity

250

PT
200
150

RI
100
50

SC
0
0 2 4 6 8 10 12

812 Concentration (mg L-1)

813
U
Fig. S3. Effect of quercetin concentration (0.05-10 mg L-1) on the RTP intensity of
AN
814 the MPA-capped Mn-doped ZnS QDs.
M

815

816
D
TE

817

818
EP

819

820
C
AC

821

822

823

824

825

826

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827 Fig. S4.

1661 1614 1388

1322
Transmittance (a.u.)

curve1

PT
1267

RI
1528 1454 1168
curve2

SC
1274
1351
1640 1581 1329 638

U
AN
2000 1600 1200 800 400

828
Wavenumber (cm-1)
M

829 Fig. S4. FT-IR spectra of quercetin and quercetin/Al3+ complexes complexes;
D

830
TE

831

832
EP

833
C

834
AC

835

836

837

838

839

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840 Fig. S5.

Average hydrodynamic
30 diameter: 24.69 nm
Frequency (%)

PT
20

10

RI
0

SC
10 15 20 25 30 35 40 45

841 Size (d. nm)

U
842 Fig. S5. Size distribution MPA-capped Mn-doped ZnS QDs (40 mg L-1) and in the
AN
843 presence of Al3+ (140 µM ).

844
M

845
D

846
TE

847

848
EP

849
C

850
AC

851

852

853

854

855

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856 Fig. S6.

A 250 B 250

200 200

150 150
△ RTP

△ RTP

PT
100 100

RI
50 50

0 0

SC
0 1 2 3 4 5 6 0 1 2 3 4 5 6

857 Concentration (mg L-1) Concentration (mg L-1)

U
858 Fig. 6. The relation ship between ∆RTP and the concentration of quercetin (from 0.15
AN
859 to 5.5 mg L-1) in Real samples (A: serum; B: urine).

860
M

861
D

862
TE

863

864
EP

865
C

866
AC

867

868

869

870

871

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872 Supplementary Table:

873

874

875

PT
876

RI
877

878

SC
879

U
880
AN
881

882
M

883
D

884
TE

885

886
EP

887
C

888
AC

889

890

891

892

893

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894 Table S1
895 Comparison of the proposed method with different analytical techniques reported for

896 detection of quercetin

Line arrange Limiting detection

Sensing matrix Ref.
(mg L-1) (mg L-1)

PT
spectrophotometric 1.0-12.0 0.76 [29]

Raman spectroscopy 15.1-45.3 15.1 [31]

RI
Fluorescence method (quercetin-
0.02-0.80 0.005 [41]
Al(III) CIP membrane)

SC
Flow injection chemiluminescence 0.42-48.3 0.28 [50]

Ionic liquids-based monolithic

0.5-10 0.1 [51]
cartridge

Electro analytical nanosensor

U
0.003-1.69 0.001 [52]
AN
Pt-PDA@SiO2/GCE 0.016-0.13 0.005 [53]

Mn-doped ZnS Quantum Dots 0.1-6.0 0.047 This work

M

897
D

898
TE

899

900
EP

901
C

902
AC

903

904

905

906

907

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908 Table S2

909 Tolerance of foreign substances.

Variation of the
Co-existing substances Tolerance ratios
calculated value (%)
NaCl 500 -3.4
KCl 500 -3.2

PT
CaCl2 150 +4.1
Glucose 500 -2.5

RI
BSA 150 +3.4
DNA 300 -2.6
L-Cys 400 +2.9

SC
L-Arg 140 +4.6
L-Gly 400 +2.7
catechin 70 -3.8

U
910
AN
911

912
M

913
D

914
TE

915

916
EP

917
C

918
AC

919

920

921

922

923

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924 Table S3

925 Intra-day detection of quercetin by MPA-capped Mn-doped ZnS QDs-Al3+ system

Mean assayed concentration

Added (mg L-1) RSD (%)
(mgL-1)
2.0 2.03 4.6

PT
4.0 4.02 3.9

926

RI
927

SC
928

929

U
AN
930

931
M

932
D

933
TE

934

935
EP

936
C

937
AC

938

939

940

941

942

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943 Table S4

944 Inter-day detection of quercetin by MPA-capped Mn-doped ZnS QDs-Al3+ system

Mean assayed concentration

Added (mgL-1) RSD (%)
(mgL-1)
2.0 2.04 4.7

PT
4.0 4.04 4.8

945

RI
U SC
AN
M
D
TE
C EP
AC

49