Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in <italic style="font-style: italic">Bombyx Batryticatus</italic> Based on UPLC－UV－Q－TOF MS

SHAO You-xin; SHANG Qiang; ZHENG Wen-lin; LI Dong; GUO Lun; HU Ping; ZHANG Hong-yang

doi:10.19969/j.fxcsxb.23051705

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Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in Bombyx Batryticatus Based on UPLC－UV－Q－TOF MS

Scientific Papers | 更新时间：2023-10-24

- Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in Bombyx Batryticatus Based on UPLC－UV－Q－TOF MS
- Journal of Instrumental Analysis Vol. 42, Issue 8, Pages: 992-999(2023)
- 作者机构：
  
  1.华东理工大学化学与分子工程学院，上海 200237
  2.四川光大制药有限公司，四川彭州 611930
  3.四川省抗病毒中药产业化工程技术研究中心，四川彭州 611930
  4.国家中药现代化工程技术研究中心川药资源分中心，四川彭州 611930
- 作者简介：
- 基金信息：
- DOI：10.19969/j.fxcsxb.23051705
  CLC：
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邵尤欣,尚强,郑文林等.基于UPLC－UV－Q－TOF MS的僵蚕嘌呤（核苷）类成分多指标定量指纹图谱研究[J].分析测试学报,2023,42(08):992-999.

SHAO You-xin,SHANG Qiang,ZHENG Wen-lin,et al.Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in Bombyx Batryticatus Based on UPLC－UV－Q－TOF MS[J].Journal of Instrumental Analysis,2023,42(08):992-999.
邵尤欣,尚强,郑文林等.基于UPLC－UV－Q－TOF MS的僵蚕嘌呤（核苷）类成分多指标定量指纹图谱研究[J].分析测试学报,2023,42(08):992-999. DOI： 10.19969/j.fxcsxb.23051705.

SHAO You-xin,SHANG Qiang,ZHENG Wen-lin,et al.Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in Bombyx Batryticatus Based on UPLC－UV－Q－TOF MS[J].Journal of Instrumental Analysis,2023,42(08):992-999. DOI： 10.19969/j.fxcsxb.23051705.

摘要

嘌呤（核苷）是机体细胞生命活动中的主要遗传物质，但目前缺乏对僵蚕中该类成分的整体定量研究。该文采用ACE Excel 2 AQ水性柱（2.1 mm × 150 mm，2 µm）进行分离，以10 mmol/L磷酸二氢钾水溶液－甲醇为流动相，分别优化了样品提取与色谱条件，建立了中药僵蚕中嘌呤（核苷）类成分的超高效液相色谱－紫外（UPLC－UV）指纹图谱方法。利用四极杆－飞行时间质谱（Q－TOF MS）对9个特征峰物质进行二级鉴定，并对其质谱裂解规律进行总结。其中，2′-甲氧基-鸟苷单磷酸、6-（,β,-羟基乙基氨基）-嘌呤、2′-甲氧基-腺苷单磷酸和N6-（2-羟乙基）-腺苷首次从僵蚕中鉴定。基于该方法，对其中的鸟嘌呤、尿苷、腺嘌呤、鸟苷、6-（,β,-羟基乙基氨基）-嘌呤、腺苷、N6-（2-羟乙基）-腺苷7个成分进行绝对定量测定。2′-甲氧基-鸟苷单磷酸和2′-甲氧基-腺苷单磷酸因缺乏对照品，采用相对定量分析。结果表明，各指标成分的精密度、重现性、稳定性、耐用性的相对标准偏差（RSD）均不大于5.0%，7种成分的线性关系良好（,r,2, ≥ 0.999 4），检出限（LOD）为0.266 ~ 0.474 μg/mL，定量下限（LOQ）为0.843 ~ 1.58 μg/mL，平均加标回收率为94.0% ~ 101%，RSD不大于3.4%。采用该方法对13批不同产地僵蚕药材进行分析，发现各成分含量具有显著差异。该研究可为僵蚕的质量评价提供技术支持和科学依据。

Abstract

The comprehensively quantitative research on purine（nucleoside） constituents in ,Bombyx batryticatus ,is still deficient as they are the main genetic substances in cell life activities of organism．In this article，by using an ACE Excel 2 AQ aqueous column（2.1 mm × 150 mm，2 µm） and a mobile phase system of 10 mmol/L potassium dihydrogen phosphate aqueous solution－methanol，an ultra-performance liquid chromatography－ultraviolet（UPLC－UV） fingerprint method for purine（nucleoside） components in ,Bombyx batryticatus, was established for the first time via the optimized sample extraction and chromatographic conditions．Nine characteristic peak compounds were identified by quadrupole time-of-flight mass spectrometry（Q－TOF MS），and the mass fragmentation patterns were summarized subsequently．Among these，2′-methoxy-guanosine monophosphate，6-（,β,-hydroxyethyl amino）-purine，2′-methoxy-adenosine monophosphate，and N6-（2-hydroxyethyl）-adenosine were firstly identified from ,Bombyx batryticatus., Seven components，including guanine，uridine，adenine，guanosine，6-（,β,-hydroxyethylamino）-purine，adenosine，and N6-（2-hydroxyethyl）-adenosine，were absolutely quantified based on the developed method．Besides，relative determinations of 2′-methoxy guanosine monophosphate and 2′-methoxy adenosine monophosphate were carried out due to the lack of standards．Method validation results demonstrated that the relative standard deviations（RSDs） for precision，reproducibility，stability and robustness of each indicator component were not more than 5.0%．Seven compounds showed good linearities（,r,2, ≥ 0.999 4），the limits of dection（LODs） were in the range of 0.266－0.474 μg/mL and the limits of quantitation（LOQs） were 0.843－1.58 μg/mL. The average spiked recoveries ranged from 94.0% to 101% with RSDs not more than 3.4%．Comparative analysis on 13 batches of ,Bombyx batryticatus, originated from different regions was then conducted，and significant differences were found in the contents of each component．This study could provide a technical support and a scientific basis for the quality evaluation on ,Bombyx batryticatus,.

关键词

僵蚕嘌呤（核苷）指纹图谱多指标定量UPLC－UV－Q－TOF MS

Keywords

Bombyx batryticatuspurine（nucleoside）fingerprintmulti-index quantificationUPLC－UV－Q－TOF MS

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Multi-index Quantitative Fingerprint Analysis of Purine （Nucleoside） Components in Bombyx Batryticatus Based on UPLC－UV－Q－TOF MS

DOI：10.19969/j.fxcsxb.23051705

摘要

Abstract

关键词

Keywords

references