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Chinese Journal of Chromatography
2010, Vol. 28, No. 6
Online: 28 June 2010

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Prediction of peptide retention time in reversed-phase liquid chromatography and its application in protein identification LIU Chao, WANG Haipeng, FU Yan, YUAN Zuofei, CHI Hao, WANG Leheng, SUN Ruixiang, HE Simin 2010, 28 (6):  529-534.  DOI: 10.3724/SP.J.1123.2010.00529 Abstract ( 3605 )   [Full Text(HTML)] () PDF (181KB) ( 437 )   Liquid chromatography-mass spectrometry (LC-MS) is the mainstream of high-throughput protein identification technology. Peptide retention time in reversed-phase liquid chromatography (RPLC) is mainly determined by the physicochemical properties of the peptide and the LC conditions (stationary phase and mobile phase). Retention time can be predicted by analyzing these properties and quantifying their effects on peptide chromatographic behavior. Prediction of peptide retention time in LC can be used to improve identification of peptides and post translational modifications (PTM). There are mainly two methods to predict retention time: i.e. retention coefficients and machine learning. The coefficient of determination between observed and predicted retention times can reach 0.93. With the development of LC-MS technology, retention time prediction will become an important tool to facilitate protein identification.

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Renaturation with simultaneous purification of the recombinant human Flt3 ligand from inclusion bodies by high performance hydrophobic interaction chromatography JIA Jia, WANG Lili, GAO Dong, GENG Xindu 2010, 28 (6):  535-540.  DOI: 10.3724/SP.J.1123.2010.00535 Abstract ( 2673 )   [Full Text(HTML)] () PDF (226KB) ( 642 )   Flt3 ligand (FL) is a class of cytokines with the functions of promoting early hematopoiesis. It has important clinical value in promoting growth and development of hematopoietic cells and hematopoietic mobilization. In order to obtain large quantities of recombinant human FL (rhFL) by genetic engineering methods for clinic and research, in this work, rhFL was expressed in E. coli as inclusion bodies. The inclusion bodies were recovered, cleaned and solubilized in 8 mol/L urea, the solubilized rhFL was renatured by high performance hydrophobic interaction chromatography (HPHIC) with simultaneous purification, the retention feature and renaturation regularity were studied. The results showed that when the denatured protein concentration was 8.51 g/L, and the end group of stationary phase was PEG800, under the conditions of mobile phase of pH 7.0 and with the addition of 4 mol/L urea, 1.8 mmol/L glutathione (GSH) and 0.3 mmol/L oxidative glutathione (GSSG), a mass recovery of 36.9% and a purity of 94.5% were obtained after refolding with simultaneous purification. The obtained rhFL was successfully renatured with simultaneous purification in only one step of HPHIC, and it provided a foundation for the manufacturing of high quality rhFL.

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General retention time formulae for gradient liquid chromatography with any combination of isocratic, linear and stepwise gradients HAO Weiqiang, DI Bin, YANG Yongbing, CHEN Qiang, WANG Junde 2010, 28 (6):  541-546.  DOI: 10.3724/SP.J.1123.2010.00541 Abstract ( 2754 )   [Full Text(HTML)] () PDF (210KB) ( 712 )   The ideal model of liquid chromatography in gradient elution was solved by using the characteristic approach on the assumption of a linear solvent strength model. By considering the influence of the dwelling time of the chromatographic system on retention time, the retention time formulae suited for any combination of isocratic, linear and stepwise gradients were obtained. It was found that the values of the retention time obtained from these formulae were in well accordance with those computed by using the finite-difference method, which confirms the validity of the formulae. Due to the simplicity and broad scope of applications, these formulae can be easily applied in practice.

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Classification of diabetes deficiency syndromes based on plasma fatty acid metabolic profilings using pre-column derivatization quantitative method XU Wenjuan, HUANG Yuhong, WANG Longxing, YANG Qianxu, XIAO Hongbin, ZHANG Deqin 2010, 28 (6):  547-550.  DOI: 10.3724/SP.J.1123.2010.00547 Abstract ( 2734 )   [Full Text(HTML)] () PDF (156KB) ( 556 )   A simple metabolic profiling approach for quantitative analysis of free fatty acids (FFAs) in human plasma by high performance liquid chromatography was described and validated, using α-bromoacetophenone as the derivative reagent and heptadecanoic acid (C17:0) as the internal standard. The quantitations of 6 predominant FFAs and 6 trace FFAs were achieved. Plasma fatty acid metabolic profiling of 75 diabetic patients was investigated, and then analyzed by multivariate statistical analysis. The linear discriminant analysis (LDA) model was established and validated for the pattern discrimination between Qi-deficiency and Qi and Yin-deficiency, with the hit ratio 94.3%. Stepwise discriminant analysis (SDA) model indicated that arachidonic acid (C20:4) and oleic acid (C18:1) contained the important information on the two syndromes above, and can be used as potential biomarkers of traditional Chinese medicine (TCM) syndrome. It is of great significance to systematically study the relationship between fatty acid metabolic profiling and TCM syndrome using metabolomics methods, and to improve the credibility and repeatability of clinical diagnosis and treatment system.

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Monodisperse thermo-sensitive chromatographic stationary phase prepared by atom transfer radical polymerization and its chromatographic properties OUYANG Kanglong, CAO ying, WANG Fuqiang, GONG Bolin 2010, 28 (6):  551-555.  DOI: 10.3724/SP.J.1123.2010.00551 Abstract ( 2674 )   [Full Text(HTML)] () PDF (200KB) ( 497 )   The new monodisperse thermo-sensitive stationary phase was prepared by atom transfer radical polymerization method at room temperature using α-bromo-isobutyryl bromide as an initiator, and the organic metal compound formed in the CuCl/CuCl2/Bpy system as a catalyst. The stationary phase was characterized by means of elementary analysis and Fourier transform infrared (FTIR) spectroscopy and evaluated in detail to determine its separability, thermosensitivity, stability and reproducibility. The grafting yield of the N-isopropylacrylamide monomer on the surface of poly(glycidyl methacrylate-co-ethylene dimethacrylate) (PGMA/EDMA) beads is 10.4%, and the stationary phase can effectively separate 4-hydroxy benzaldehyde, o-cresol and 4-n-butylaniline by changing temperature. The results showed that the synthesized stationary phase has satisfactory chromatographic properties, thermo-sensitive properties and reproducibility.

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Determination of imidazolium ionic liquid cations by ion-pair chromatography using gradient elution GAO Wei, YU Hong, MA Yajie 2010, 28 (6):  556-560.  DOI: 10.3724/SP.J.1123.2010.00556 Abstract ( 2733 )   [Full Text(HTML)] () PDF (176KB) ( 536 )   Determination of five imidazolium ionic liquid cations by ion-pair chromatography was carried out using ultraviolet-visible spectrophotometry (UV/VIS)detection. Chromatographic separations were performed on a ZORBAX Eclipse XDB-C18 column with 1-heptanesulfonic acid sodium (adjusted by citric acid to pH 4.0)+acetonitrile as eluent. The effects of ion-pair reagents, acetonitrile concentration and column temperature on retention of the cations were investigated. The retention factor of imidazolium cation on column was increased with the increase of the concentration of 1-heptanesulfonic acid sodium. With the increase of the concentration of acetonitrile, the retention time of imidazolium cation was obviously shortened and the peak shape was improved. Under the optimum conditions, detection limits (S/N=3) for the cations were 0.05-0.30 mg/L. Relative standard deviations (RSDs) for peak areas were less than 0.1%. The method has been successfully applied to the determination of two ionic liquids by chemically synthesized in laboratory with the recoveries of the cations of 98.6%-102.1% after spiking.

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Quantitative determination of Cantide, an antisense oligodeoxynucleotide in rhesus monkey plasma using non-gel sieving capillary electrophoresis method WANG Xiuzhong, WANG Qingqing, WANG Shihong, LI Weiping, SONG Haifeng, LU Dandan, WANG Shengqi 2010, 28 (6):  561-565.  DOI: 10.3724/SP.J.1123.2010.00561 Abstract ( 2886 )   [Full Text(HTML)] () PDF (177KB) ( 499 )   A dual solid phase extraction (SPE) pretreatment coupling with non-gel sieving capillary electrophoresis (NGCE) analysis method was established for the quantitative determination of an antisense oligodeoxynucleotide, Cantide, in rhesus monkey plasma. The conditions of SPE and the NGCE analysis were optimized. Under the optimized conditions (the SPE conditions: the pH of loading buffer was 9.0; the volumes of loading and the elution solution for the anion-exchange column were 5 mL and 3 mL, respectively. The NGCE analysis conditions: loading gel time was 30 min and the separation voltage was 24 kV), the linear dynamic range of Cantide in rhesus monkeys plasma was 1.95-250 mg/L, and the correlation coefficient (r) was more than 0.998. The limit of quantitation was 1.95 mg/L. The intra-batch accuracies ranged from 93.38% to 100.71% with the intra-batch relative standard deviation (RSD) less than 11%. The inter-batch accuracies were from 89.46% to 103.46% with the inter-batch RSD less than 9%. The stability experiment showed that the Cantide plasma sample was stable when stored at 4 ℃ for 24 h, room temperature for 4 h, -80 ℃ for 30 days and freeze-thaw for 2 cycles. This method was finally successfully applied to pharmacokinetic study of Cantide in rhesus monkeys.

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Comparison of phenolic components among different species of Dendrobium (Shihu Fengdou) and determination of their active components—moscatilin and gigantol ZHOU Jing, XU Zhiliang, KONG Hongwei, LU Xin, XU Guowang 2010, 28 (6):  566-571.  DOI: 10.3724/SP.J.1123.2010.00566 Abstract ( 2586 )   [Full Text(HTML)] () PDF (332KB) ( 811 )   A liquid chromatography coupled with ion trap-time of flight mass spectrometry (LC-IT-TOF MS/MS) profiling method was developed for the analysis of the phenolic constituents in Dendrobium. Six different species of Dendrobium were analyzed. Eighteen phenolic constituents in the samples were tentatively identified based on their mass spectra and related literatures. Moscatilin and gigantol were chosen as potential quality control targets of all species of Dendrobium. A high performance liquid chromatography (HPLC) method was developed for quantitative analysis of moscatilin and gigantol in Dendrobium. The extraction condition for moscatilin and gigantol was optimized by the response surface methodology (RSM). The linearity of the method is more than 0.9998 and the precision is within 3% for all the two analytes. The limits of detection (LOD) were 0.18 mg/L and 0.09 mg/L respectively. Recoveries were 97.1% and 101.4% respectively. The study might provide a basis for the quality control of Dendrobium.

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Determination of β2-agonists and β-blockers in urine using high performance liquid chromatography-ion trap mass spectrometry MIAO Hong, ZOU Jianhong, FAN Sai, GAN Lewen, ZHAO Yunfeng, WU Yongning, 2010, 28 (6):  572-578.  DOI: 10.3724/SP.J.1123.2010.00572 Abstract ( 2706 )   [Full Text(HTML)] () PDF (277KB) ( 523 )   A method has been developed for the determination of 23 β2-agonists and 5 β-blockers in urine samples using high performance liquid chromatography-ion trap mass spectrometry (HPLC-IT-MS). Urine samples were first deproteinized by high-speed frozen centrifugation, and the supernatants were loaded on an ExtrelutTM diatomite column for clean-up. The analytes were eluted by ethyl acetate and concentrated for further analysis. The analytical separation was performed on an AtlantisT3-150 mm chromatographic column with the gradient elution using methanol and water (containing 0.1% formic acid). The detection was carried on a linear ion trap mass spectrometer under multiple reaction monitoring (MRM) mode with the source operated in positive mode of electrospray ionization (ESI+). Nine deuterium labeled β2-agonists were used as internal standards for quantitative analysis. The results showed that the linear ranges for 23 β2-agonists and 5 β-blockers were 0.005－0.16 mg/L, and the limits of detection were all around 0.2 μg/L. The mixed standard solution was added into the blank urine samples, and the recoveries of 23 β2-agonists and 5 β-blockers were ranged from 57.1% to 127.7% with the relative standard deviations of 1.1%-31.1%. The results demonstrate that the method is easy, fast, sensitive, and suitable for the confirmation and quantification of 23 β2-agonists and 5 β-blockers in urine samples.

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Determination of 215 pesticide residues in ginger using liquid chromatography coupled with electrospray ionization tandem mass spectrometry CAO Jing, PANG Guofang, WANG Minglin, FAN Chunlin 2010, 28 (6):  579-589.  DOI: 10.3724/SP.J.1123.2010.00579 Abstract ( 2615 )   [Full Text(HTML)] () PDF (364KB) ( 1070 )   A multiresidue analytical method was developed for the determination of 215 pesticides in ginger using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The pesticide residues were extracted from ginger by acetonitrile containing 1% (v/v) acetic acid, cleaned-up by a Sep-Pak Vac cartridge, eluted with acetonitrile-toluene (3:1, v/v). The eluate was concentrated to about 0.5 mL with a rotary evaporator, dried with nitrogen at room temperature. The sample was redissolved in an acetonitrile-water mixture (3:2, v/v), then analyzed using LC-MS/MS in multiple reaction monitoring (MRM) mode via positive electrospray ionization. The recovery test was conducted at spiked level of limit of quantification (LOQ). The validation results were as follows: the overall recoveries were from 68.1% to 132.6% of which 94.4% of the recoveries were from 70% to 120%, with the relative standard deviations of 0.4%-25.0%. The limits of detection (S/N=3) and the limits of quantification (S/N=10) were 0.01-70.45 μg/L and 0.04-234.84 μg/L, respectively. The results demonstrated that this method is simple and with acceptable sensitivity and accuracy to meet the requirements of the multiple pesticide residue analysis. This method is applicable to determine 215 pesticide residues in ginger.

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Simultaneous determination of 7 rhodamine dyes in hot chili products by high performance liquid chromatography-tandem mass spectrometry HU Xia, XIAO Guang, PAN Wei, MAO Xiqin, LI Peng 2010, 28 (6):  590-595.  DOI: 10.3724/SP.J.1123.2010.00590 Abstract ( 2399 )   [Full Text(HTML)] () PDF (212KB) ( 610 )   A credible method was developed for the simultaneous determination of 7 rhodamine dyes in hot chili products based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The samples were extracted with hexane or methanol-water (1:1, v/v) and then cleaned up by a solid phase extraction cartridge. The target analytes were separated on an SB-C18 column with gradient elution using acetonitrile and water (containing 0.1% (v/v) formic acid for both) as mobile phases. The identification and quantification were achieved by using ESI-MS/MS in positive ion mode and with multiple reaction monitoring (MRM). The linear ranges were from 0.0005 to 1.0 mg/L with the correlation coefficients (r2) above 0.997 for all the 7 rhodamine dyes. The limits of detection (LOD) in chili powder and chili oil were from 0.21 to 51 μg/kg and 0.19 to 25 μg/kg, respectively. The relative standard deviations of intra-day and inter-day were both less than 20%. The recoveries of the method were between 85.0% and 106%. The method is simple, rapid, highly sensitive and suitable for the simultaneous determination of 7 rhodamine dyes in foods.

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Rapid determination of 3 nitromidazole residues in honey using liquid chromatography-tandem mass spectrometry LIU Yongming, CAO Yanzhong, LI Jin, WU Yanping, ZHANG Jinjie, LI Xuemin, GE Na 2010, 28 (6):  596-600.  DOI: 10.3724/SP.J.1123.2010.00596 Abstract ( 3037 )   [Full Text(HTML)] () PDF (176KB) ( 550 )   A method was developed for the rapid determination of 3 nitromidazole residues in honey, including metronidazole, ronidazole and dimetridazole, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The honey sample was dissolved in water, and the solution was cleaned up with an Oasis HLB cartridge. The cartridge was washed with water and methanol-water solution (1:9, v/v) in turn, and eluted with ethyl acetate. The solution was concentrated and then analyzed by LC-MS/MS. External calibration was used for quantitative determination. The recoveries and relative standard deviations (n=8) were from 76.6% to 89.7% and 5.2% to 9.9%, at the spiked levels of 0.05-2.0 μg/kg, respectively. The limits of detection were 0.1 μg/kg for metronidazole, and 0.2 μg/kg for ronidazole and dimetridazole. The method was successfully applied for the inspection of exported honey. With simple and fast sample preparation, the method is sensitive and specific for the determination. The sensitivity and accuracy of the method meet the requirements of the inspection for the 3 nitromidazole residues in honey in Japan and European Union.

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Determination of zearalenone and related mycotoxins in grain and its products by solid-phase extraction coupled with ultra performance liquid chromatography-tandem mass spectrometry MENG Juan, ZHANG Jing, ZHANG Nan, SHI Jiachen, SHAO Bing 2010, 28 (6):  601-607.  DOI: 10.3724/SP.J.1123.2010.00601 Abstract ( 3355 )   [Full Text(HTML)] () PDF (257KB) ( 925 )   A method was established for the determination of 6 zearalenonic compounds (α-zearalanol, β-zearalanol, α-zearalenol, β-zearalenol, zearalanone and zearalenone) in grain and its products based on solid-phase extraction coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples was extracted by 84% (v/v) acetonitrile-water solution and further purified by an ENVI-Carb graphite carbon black (GCB) cartridge, which was eluted by 6 mL dichloromethane-methanol (7:3, v/v) solution. The target compounds were assayed by UPLC-MS/MS. The chromatographic separation was performed on an ACQUITY UPLCTM BEH C18 column with gradient elution using acetonitrile and water as mobile phases. The mass spectrometric acquisitions were carried out by means of multiple reaction monitoring (MRM) in electrospray negative ionization mode. The good linearities (R2＞0.99) were achieved for the 6 compounds over the range of 0.1-50 μg/L based on the internal standard calibration of α-zearalenol-d4. The detection limits of the method were 0.1-0.2 μg/kg. The mean recoveries of the 6 target compounds (spiked at three concentration levels) ranged from 79.9% to 104.0%, with the relative standard deviations (RSDs) no more than 10%. It has been applied in the analysis of grain and related products taken from Beijing. As a result, zearalenone presented a highest detectable frequency, with a concentration range of 0.42-220.7 μg/kg. In addition, α-zearalenol and β-zearalenol were also detected in this survey. This proposed method is simple, sensitive, reproducible, and complied with the regulations for the determination of trace contaminants residues in food matrices.

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Analysis of volatile and semi-volatile compounds in tobacco using off-line combination of liquid chromatography and capillary gas chromatography-mass spectrometry BAI Junchao, LIU Shaofeng, XIE Fuwei, LIU Huimin, XIA Qiaoling 2010, 28 (6):  608-614.  DOI: 10.3724/SP.J.1123.2010.00608 Abstract ( 3026 )   [Full Text(HTML)] () PDF (275KB) ( 592 )   A method of off-line combination of liquid chromatography and capillary gas chromatography-mass spectrometry (LC-CGC/MS) was established for the analysis of volatile and semi-volatile compounds in tobacco. The separation mechanism of volatile and semi-volatile compounds was studied. An LC column of 250 mm×2.0 mm packed with 5 μm amino-bonded silica was used as stationary phase for the separation. n-Hexane/dichloromethane/acetonitrile (90:6.6:3.4, v/v/v) was used as mobile phase. Five fractions from one LC run were collected in five nitrogen-blowing flasks, separately. The same fractions from 10 LC runs were combined together and concentrated to 1 mL by nitrogen blowing, and then were analyzed by CGC/MS. The column of DB-5MS (60 m×0.25 mm×0.25 μm) was used for CGC/MS analysis. Comparing with direct CGC/MS analysis of the same sample, the LC-CGC/MS system is suitable for a complex sample, and has a higher reliability of qualitative analysis.

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Simultaneous analysis of twenty free amino acids in tobacco using liquid chromatography-electrospray ionization/ion trap tandem mass spectrometry HUANG Yifei, HU Jing 2010, 28 (6):  615-622.  DOI: 10.3724/SP.J.1123.2010.00615 Abstract ( 2748 )   [Full Text(HTML)] () PDF (280KB) ( 785 )   A method for simultaneous analysis of 20 free amino acids in tobacco was developed using liquid chromatography-electrospray ionization/ion trap tandem mass spectrometry (LC-ESI-IT-MS/MS). Tobacco samples were extracted and filtered, and analyzed without derivatization or solid phase extraction. The LC separation was carried on a reversed-phase HyPURITY C18 column (200 mm×2.1 mm, 5 μm) with the mobile phase of 1%(v/v) acetonitrile/water solution containing 0.1% nonafluoropentanoic acid and 90% acetonitrile/water solution containing 0.1% nonafluoropentanoic acid in gradient mode. The results showed that the limits of detection (LODs) of 20 amino acids were in the range of 0.01-0.05 μmol/L (S/N=3), the linear correlation coefficients were above 0.9977 in all cases, and the relative standard deviations (RSDs) of peak areas of the extracted ion chromatograms were in the range of 0.78%-4.93%. The developed method is of good efficiency, sensitivity and selectivity, and has been successfully applied to determine the free amino acids in varieties of tobacco samples.

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Modification of mid-point restrictor of heart cut multidimensionalgas chromatography-mass spectrometry system and its application in analysis of benzo［a］pyrene in cigarette smoke SHI Jiaqin, LIU Baizhan, XIE Wenyan 2010, 28 (6):  623-627.  DOI: 10.3724/SP.J.1123.2010.00623 Abstract ( 2309 )   [Full Text(HTML)] () PDF (216KB) ( 594 )   The mid-point restrictor, known as the key spare part of multidimensional gas chromatography (MDGC), was modified and applied in the determination of benzo［a］pyrene in cigarette smoke. The graphite restrictor was replaced by a 0.25 mm i.d. fused silica open tubular and the two-hole graphite ferrule was replaced by a press-fit glass Y-splitter to improve the stability of retention time on the 1st dimensional column and eliminate the carry-over effects of graphite. The cigarette samples were prepared as follows: The mainstream cigarette smoke collected on Cambridge pad was extracted with cyclohexane, then the extractant was analyzed by heart cut MDGC-mass spectrometry(MS) directly with benzo［a］pyrene-d12 as internal standard. The method was proved to be simple, sensitive, fast and reliable with good linearity (linear range of 1.47-29.4 μg/L, r2 of 0.9999), reproducibility (relative standard deviation (RSD) of 1.94%), recovery (90.74%-101.86%) and low quantitative and qualitative detection limits. Moreover, the amount of benzo［a］pyrene in the 2R4F Kentucky reference cigarette smoke obtained by this method was close to that reported in literature.

Technical Notes

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Determination of acrylic acid from catalytic preparation lactic acid by anion-exchange chromatography SHI Haining, WANG Hui, TAO Lizhi, WANG Zonghua, DING Mingyu 2010, 28 (6):  628-631.  DOI: 10.3724/SP.J.1123.2010.00628 Abstract ( 3199 )   [Full Text(HTML)] () PDF (125KB) ( 593 )   Acrylic acid is a kind of important monomer and basic organic chemical raw material. In the process of catalytic preparation of acrylic acid from lactic acid, in order to monitor the catalytic process effectively and timely, an anion-exchange chromatographic (AEC) method has been established for the simultaneous determination of lactic acid and acrylic acid. The separation was carried out on a Metrohm A Supp 5 anion-exchange column (150 mm×4.0 mm) with 2 mmol/L Na2CO3+2 mmol/L NaHCO3 as the mobile phase. The flow rate of the mobile phase was 0.7 mL/min. A chemically suppressed conductivity detector was used. The linear ranges of calibration curves were 0.1-500 mg/L for lactic acid and 0.1-200 mg/L for acrylic acid. The detection limits with S/N=3 were 0.030 mg/L for lactic acid and 0.035 mg/L for acrylic acid. The recoveries of lactic acid and acrylic acid were 100.7%-106% and 99.6%-103% with the relative standard deviations of 2.16%-2.49% and 2.42%-2.48%, respectively. This method is accurate, speedy, sensitive and reproducible, and has been successfully used for the determination of lactic acid and acrylic acid in the catalytic reaction product.

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Determination of arbutin in apple juice concentrate by ultra performance liquid chromatography with electrospray ionization tandem mass spectrometry KONG Xianghong, HE Qiang, YUE Aishan, WU Shuangmin, LI Jianhua 2010, 28 (6):  632-634.  DOI: 10.3724/SP.J.1123.2010.00632 Abstract ( 2622 )   [Full Text(HTML)] () PDF (123KB) ( 567 )   An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of arbutin in apple juice concentrate. Samples were diluted with water, then cleaned-up with a PS-DVB column. Quantitation was carried out using an external standard method. UPLC was performed on an Eclipse Plus C18 column(100 mm×2.1 mm, 1.8 μm) using a gradient solvent system (methanol-water). MS/MS was performed with multiple reaction monitoring (MRM) mode. The detection limit of arbutin was 0.02 mg/L. The method showed good linear relationship at the range of 0.04-2.0 mg/L. The recoveries ranged from 75.2% to 102.7% with relative standard deviations (RSDs) less than 8.9%. The method is simple, fast and sensitive. It’s suitable for quantitative and qualitative analysis of arbutin in apple juice concentrate.