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| Determination of Choline and Acetylcholine in Honey by Ultra Performance Liquid Chromatography－Tandem Mass Spectrometry |
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| DOI： |
| KeyWord:choline acetylcholine honey ultra performance liquid chromatography－tandem mass spectrometry（UPLC－MS/MS） |
| Author | Institution |
| XUE Xia，DING Yi，GONG Pi-xue，LU Lan-xiang，WANG Jun，ZHANG Yan-xia，WU Chuan-xiang，SU Shu-fang，WEI Li-li，LIU Yan-ming，HU Mei |
Shandong Institute for Food and Drug Control，Shandong Research Center of Engineering and Technology for Quality Control of Food for Special Medical Purposes，Shandong Research Center of Engineering and Technology for Safety Inspection of Food and Drug，Jinan ，China |
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| Abstract: |
| An analytical method was developed for the determination of choline and acetylcholine in honey using ultra performance liquid chromatography－tandem mass spectrometry（UPLC－MS/MS）．The samples were firstly extracted with water－acetonitrile solution（2∶3，by volume），then diluted with acetonitrile－water solution（8∶2） and filtered through a 0.22 μm microporous membrance，and finally determined using the UPLC－MS/MS system．The choline and acetylcholine were separated on an ACQUITY UPLC BEH HILIC column（100 mm × 2.1 mm，1.7 μm）by gradient elution，using 10 mmol/L ammonium acetate solution containing 0.1% acetic acid and acetonitrile as mobile phases．The target compounds were analyzed with electrospray ion source in positive electrospray ionization mode under the multi-reaction monitoring（MRM） mode，and quantified by the external standard method．Results showed that，under the optimal conditions，choline and acetylcholine showed good linearities（r2 > 0.99） in the ranges of 0.5－50 ng/mL and 0.05－5 ng/mL，while the limits of detection（LODs，S/N = 3） for them were 0.5 mg/kg and 0.05 mg/kg，and the limits of quantification（LOQs，S/N = 10） were 1.0 mg/kg and 0.1 mg/kg，respectively．The recoveries for choline and acetylcholine in blank high fructose syrup at three levels of LOQs，10LOQs and 50LOQs ranged from 100% to 103%，with relative standard deviations（RSDs） less than 5.0%，while the recoveries for the analytes in a honey sample at the spiked level of 50LOQs were 97.3% and 97.8%，respectively，with RSDs not more than 5.1%．With the advantages of convenience，sensitivity，good recoveries and repeatability，the method was suitable for the detection of choline and acetylcholine in honey．This method was successfully applied to the analysis on 70 batches of honey and 10 batches of syrup． |
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