Rapid Communications in Mass Spectrometry最新文献_第7页

Software-assisted automated detection and identification of “unknown” analogues of benzodiazepines in liquid chromatography mass spectrometry analysis 软件辅助自动检测和识别液相色谱质谱分析中的苯并二氮杂卓 "未知 "类似物。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-08-05 DOI: 10.1002/rcm.9883

Dana Marder, Ori Gutman, Uriel Bretler, Yiffat Katz, Lilach Yishai-Aviram, Eyal Drug

Rationale

Benzodiazepines (BZDs) construct a large group of psychoactive drugs acting as depressants of the central nervous system (CNS) and used in medicine as sedatives and anxiolytic and antiepileptic agents. The illicit use of these materials is a worldwide problem, and for many years, part of the benzodiazepines have been abused as rape drugs. For example, flunitrazepam (Rohypnol) is most commonly linked by media reports to drug-facilitated sexual assaults, more commonly referred to as “date rape.” Furthermore, there are growing concerns for other misuses of these drugs. Over the last few years, there was an increase in the number, type, and availability of new psychoactive substances (NPS) belonging to the benzodiazepine group, challenging standard forensic labs to fully identify the chemical structure of new, unknown benzodiazepines.

Methods

This work demonstrates a new application of the automated tool for the detection and identification of benzodiazepine analogues using high-resolution-accurate-mass LC-MS analysis, followed by “Compound Discoverer” (CD) software data processing, to automatically detect various benzodiazepine analogues by picking peaks and compare them to in silico calculated modifications made on a predefined basic backbone. Subsequently, a complete structural elucidation for the proposed molecular formula is obtained by MS/MS data analysis of the suspected component.

Results

This method was found to be useful for the automated detection and putative identification of a series of nine “unknown” benzodiazepine analogues, at concentrations in the low ng/mL range.

Conclusions

We hereby present a general demonstration of this powerful tool for the forensic community in the detection and identification of hazardous unknown compounds.

理由：苯二氮卓类（BZDs）是一大类精神活性药物，具有抑制中枢神经系统（CNS）的作用，在医学上用作镇静剂、抗焦虑剂和抗癫痫剂。非法使用这类药物是一个世界性问题，多年来，部分苯二氮卓类药物被当作强奸药物滥用。例如，在媒体报道中，氟硝西泮（Rohypnol）最常与借助药物的性侵犯联系在一起，更常被称为 "约会强奸"。此外，人们越来越关注这些药物的其他滥用情况。在过去几年中，属于苯并二氮杂卓类药物的新型精神活性物质（NPS）的数量、种类和供应量都有所增加，这对标准法医实验室全面鉴定新型未知苯并二氮杂卓类药物的化学结构提出了挑战：这项工作展示了自动检测和鉴定苯并二氮杂卓类似物工具的新应用，该工具使用高分辨率-精确质量 LC-MS 分析，然后使用 "化合物发现者"（CD）软件进行数据处理，通过拾取峰值自动检测各种苯并二氮杂卓类似物，并将其与在预定义的基本骨架上进行的硅学计算修饰进行比较。随后，通过对可疑成分的 MS/MS 数据分析，对提出的分子式进行完整的结构阐释：结果：发现该方法可用于自动检测和推定鉴定一系列九种 "未知 "苯并二氮杂卓类似物，浓度在低纳克/毫升范围内：在此，我们向法医界展示了这一强大工具在检测和鉴定危险未知化合物方面的普遍应用。

{"title":"Software-assisted automated detection and identification of “unknown” analogues of benzodiazepines in liquid chromatography mass spectrometry analysis","authors":"Dana Marder, Ori Gutman, Uriel Bretler, Yiffat Katz, Lilach Yishai-Aviram, Eyal Drug","doi":"10.1002/rcm.9883","DOIUrl":"10.1002/rcm.9883","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>Benzodiazepines (BZDs) construct a large group of psychoactive drugs acting as depressants of the central nervous system (CNS) and used in medicine as sedatives and anxiolytic and antiepileptic agents. The illicit use of these materials is a worldwide problem, and for many years, part of the benzodiazepines have been abused as rape drugs. For example, flunitrazepam (Rohypnol) is most commonly linked by media reports to drug-facilitated sexual assaults, more commonly referred to as “date rape.” Furthermore, there are growing concerns for other misuses of these drugs. Over the last few years, there was an increase in the number, type, and availability of new psychoactive substances (NPS) belonging to the benzodiazepine group, challenging standard forensic labs to fully identify the chemical structure of new, unknown benzodiazepines.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>This work demonstrates a new application of the automated tool for the detection and identification of benzodiazepine analogues using high-resolution-accurate-mass LC-MS analysis, followed by “Compound Discoverer” (CD) software data processing, to automatically detect various benzodiazepine analogues by picking peaks and compare them to in silico calculated modifications made on a predefined basic backbone. Subsequently, a complete structural elucidation for the proposed molecular formula is obtained by MS/MS data analysis of the suspected component.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>This method was found to be useful for the automated detection and putative identification of a series of nine “unknown” benzodiazepine analogues, at concentrations in the low ng/mL range.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>We hereby present a general demonstration of this powerful tool for the forensic community in the detection and identification of hazardous unknown compounds.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/rcm.9883","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141892470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Ionization characteristics of various amino acids in positive-ion helium direct analysis in real-time quadrupole time-of-flight mass spectrometry 实时四极杆飞行时间质谱法正离子氦直接分析中各种氨基酸的电离特性。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-08-02 DOI: 10.1002/rcm.9879

Jianing Liu, Peng Yu, Tong Wu, Delai Ma, Yankun Sun, Xinran Zhang, Hongmei Yang

引用次数: 0

Effects of pre-treatment, historical age, and sample characteristics on the stable isotope analyses of killer whale (Orcinus orca) bone 处理前、历史年龄和样本特征对虎鲸（Orcinus orca）骨骼稳定同位素分析的影响。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-08-01 DOI: 10.1002/rcm.9874

Kelly R. Bowen, Carolyn M. Kurle

Rationale

Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone.

Methods

Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the δ¹³C and δ¹⁵N values of powdered killer whale (Orcinus orca) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics.

Results

No significant differences in the δ¹⁵N values were observed for any of the experimental protocols. However, the δ¹³C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics.

Conclusions

If only the δ¹⁵N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ¹³C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally.

理由：通过对骨骼进行稳定同位素分析，可以深入了解动物的觅食情况，并随着时间的推移进行生态重建，但要分离胶原蛋白，需要进行预处理。预处理通常包括去矿物质以去除无机成分和/或提取脂质以去除脂肪，但这些方法会因涉及的化学物质、组织和/或物种的不同而对稳定碳（δ13C）和氮（δ15N）同位素值产生不同的影响。针对不同物种的方法为各项研究的可比性提供了标准，并加深了对从现代鲸类骨骼中分离胶原蛋白的理解：方法：使用元素分析仪和同位素比质谱仪测量虎鲸（Orcinus orca）骨骼粉末的 δ13C 和 δ15N 值，这些骨骼粉末既可以是完整的（对照组），也可以在三种实验条件中选择一种：去矿物质、脂质萃取以及去矿物质和脂质萃取。此外，还对 C:N 比率进行了评估，作为胶原蛋白纯度的替代指标。最后，研究了对照组 C:N 比率与历史年龄之间的相关性，以及对照组 C:N 比率与样品特征之间的相关性：任何实验方案的 δ15N 值均无明显差异。但是，δ13C 值在所有三种实验方案中都有显著增加：脱矿、脂质提取和两种处理方法的结合。影响最大的实验方案是脱矿和脂质提取。C:N比率的测量值也因去矿物质化和两种处理方法的结合而显著降低，表明处理后的材料更接近于纯胶原蛋白。通过 C:N 比值显示的胶原蛋白纯度与历史年龄或样本特征无关：结论：如果只对虎鲸骨骼的 δ15N 值感兴趣，则无需进行预处理。如果对 δ13C 值感兴趣，则应对样本进行脱矿和脂质提取。由于历史年代和样本特征与样本污染无关，因此可以对所有样本一视同仁。

{"title":"Effects of pre-treatment, historical age, and sample characteristics on the stable isotope analyses of killer whale (Orcinus orca) bone","authors":"Kelly R. Bowen, Carolyn M. Kurle","doi":"10.1002/rcm.9874","DOIUrl":"10.1002/rcm.9874","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (<i>δ</i><sup>13</sup>C) and nitrogen (<i>δ</i><sup>15</sup>N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the <i>δ</i><sup>13</sup>C and <i>δ</i><sup>15</sup>N values of powdered killer whale (<i>Orcinus orca</i>) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>No significant differences in the <i>δ</i><sup>15</sup>N values were observed for any of the experimental protocols. However, the <i>δ</i><sup>13</sup>C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>If only the <i>δ</i><sup>15</sup>N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the <i>δ</i><sup>13</sup>C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/rcm.9874","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141873752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of the headspace gas chromatography–tandem mass spectrometry method for ethylene oxide and ethylene chlorohydrin residue in medical devices 开发顶空气相色谱-串联质谱法检测医疗器械中的环氧乙烷和乙烯氯醇残留量。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-24 DOI: 10.1002/rcm.9869

Yi Chun Chiang, Ming Chih Fang, Chuan Kai Yang, Sheng Wei Wang, Su Hsiang Tseng

Rationale

Ethylene oxide (EO) sterilization is commonly employed for the sterilization of medical devices and has a very high market share. However, EO and its metabolite ethylene chlorohydrin (ECH) are toxic to humans. In compliance with the classification and residue limits of medical devices defined by ISO 10993-7, our study established two extraction methods for the testing of EO and ECH.

Methods

The first method involves simulated-use extraction using water as the extraction solvent. While the second, exhaustive extraction, directly extracts sample through headspace sampling analysis. Gas chromatography–tandem mass spectrometry in multiple reaction monitoring mode was utilized, requiring only 16 min. Then, the developed method was applied to assess 10 commercially available medical devices sterilized by EO.

Results

In simulated-use extraction, calibration curves were evaluated in the range of 1–100 and 5–500 μg for EO and ECH, respectively (r > 0.999). Inter-day recoveries ranged from 85.0% to 95.2% and from 94.8% to 102.4%. In exhaustive extraction, calibration curves spanned 0.5–50 and 2–200 μg for EO and ECH, respectively (r > 0.999). Inter-day recoveries ranged from 101.6% to 102.1% for EO and from 98.1% to 102.2% for ECH. After analysis of the 10 commercially available medical devices, two cotton swabs were found to have ECH of 35.1 and 28.4 μg per device, and four medical devices were found to have EO with concentration below the limit of quantification. Meanwhile, we found that the EO internal standard (propylene oxide) recommended by ISO 10993-7 had interference problems with other similar substances and was not suitable as an internal standard for EO.

Conclusions

This study offers a sensitive and straightforward analytical approach to EO and ECH residues in a variety of medical devices. In addition, the results show that the EO or ECH content of these types of medical devices in our study falls below the regulatory limits, therefore instilling confidence among consumers regarding their safe use.

理由：环氧乙烷（EO）灭菌法常用于医疗器械的灭菌，市场占有率非常高。然而，环氧乙烷及其代谢物乙烯氯海德林（ECH）对人体有毒。根据 ISO 10993-7 规定的医疗器械分类和残留限量，我们的研究建立了两种提取方法来检测环氧乙烷和 ECH：第一种方法是以水为提取溶剂进行模拟使用提取。方法：第一种方法是以水为萃取溶剂进行模拟萃取，第二种方法是通过顶空取样分析直接萃取样品。采用多反应监测模式的气相色谱-串联质谱法，仅需 16 分钟。然后，将所开发的方法用于评估经环氧乙烷灭菌的 10 种市售医疗器械：在模拟使用萃取过程中，EO 和 ECH 的校准曲线范围分别为 1-100 和 5-500 μg（r > 0.999）。日间回收率分别为 85.0% 至 95.2% 和 94.8% 至 102.4%。在穷举萃取中，EO 和 ECH 的校准曲线分别为 0.5-50 和 2-200 μg（r > 0.999）。环氧乙烷的日间回收率为 101.6%至 102.1%，环己烷的日间回收率为 98.1%至 102.2%。对 10 个市售医疗器械进行分析后，发现两个棉拭子的 ECH 分别为 35.1 微克和 28.4 微克，4 个医疗器械的 EO 浓度低于定量限。同时，我们发现 ISO 10993-7 推荐的环氧乙烷内标（环氧丙烷）与其他类似物质存在干扰问题，不适合作为环氧乙烷的内标：这项研究为各种医疗器械中的环氧乙烷和环己烷残留量提供了一种灵敏而直接的分析方法。此外，研究结果表明，在我们的研究中，这些类型的医疗器械中的环氧乙烷或环己烷含量低于监管限值，因此消费者对其安全使用充满信心。

{"title":"Development of the headspace gas chromatography–tandem mass spectrometry method for ethylene oxide and ethylene chlorohydrin residue in medical devices","authors":"Yi Chun Chiang, Ming Chih Fang, Chuan Kai Yang, Sheng Wei Wang, Su Hsiang Tseng","doi":"10.1002/rcm.9869","DOIUrl":"10.1002/rcm.9869","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>Ethylene oxide (EO) sterilization is commonly employed for the sterilization of medical devices and has a very high market share. However, EO and its metabolite ethylene chlorohydrin (ECH) are toxic to humans. In compliance with the classification and residue limits of medical devices defined by ISO 10993-7, our study established two extraction methods for the testing of EO and ECH.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>The first method involves simulated-use extraction using water as the extraction solvent. While the second, exhaustive extraction, directly extracts sample through headspace sampling analysis. Gas chromatography–tandem mass spectrometry in multiple reaction monitoring mode was utilized, requiring only 16 min. Then, the developed method was applied to assess 10 commercially available medical devices sterilized by EO.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>In simulated-use extraction, calibration curves were evaluated in the range of 1–100 and 5–500 μg for EO and ECH, respectively (<i>r</i> > 0.999). Inter-day recoveries ranged from 85.0% to 95.2% and from 94.8% to 102.4%. In exhaustive extraction, calibration curves spanned 0.5–50 and 2–200 μg for EO and ECH, respectively (<i>r</i> > 0.999). Inter-day recoveries ranged from 101.6% to 102.1% for EO and from 98.1% to 102.2% for ECH. After analysis of the 10 commercially available medical devices, two cotton swabs were found to have ECH of 35.1 and 28.4 μg per device, and four medical devices were found to have EO with concentration below the limit of quantification. Meanwhile, we found that the EO internal standard (propylene oxide) recommended by ISO 10993-7 had interference problems with other similar substances and was not suitable as an internal standard for EO.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study offers a sensitive and straightforward analytical approach to EO and ECH residues in a variety of medical devices. In addition, the results show that the EO or ECH content of these types of medical devices in our study falls below the regulatory limits, therefore instilling confidence among consumers regarding their safe use.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Conversion of 5α-Androstane-3α,17β-diol to bis[(4-dimethylamino)phenyl carbamate] derivative for sensitive determination of its rat brain level by LC/ESI-MS/MS 通过 LC/ESI-MS/MS 将 5α-雄甾烷-3α,17β-二醇转化为双[(4-二甲基氨基)苯基氨基甲酸酯]衍生物，以灵敏测定其在大鼠大脑中的含量。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-24 DOI: 10.1002/rcm.9875

Tatsuya Higashi, Asuka Tanaka, Shiho Tsubura, Shoichi Nishimoto-Kusunose

Rationale

5α-Androstane-3α,17β-diol (3α,5α-Adiol) is a testosterone-derived neurosteroid and has anxiolytic and analgesic effects via γ-aminobutyric acid type A receptors as with the progesterone-derived neurosteroid, allopregnanolone (AP). Although the psychotropic drug-evoked changes in the brain AP concentration have been intensively studied, those in the brain 3α,5α-Adiol concentration remain poorly understood. One of the causes for this is the limited availability of a validated method for quantifying the brain 3α,5α-Adiol with a sufficient sensitivity and specificity, which is described in this study.

Methods

To enhance the detectability of 3α,5α-Adiol by electrospray ionization-tandem mass spectrometry (ESI-MS/MS), derivatization with 4-dimethylaminobenzoyl azide was employed. The brain sample was purified by solid-phase extraction and the recovered 3α,5α-Adiol and the deuterated internal standard were derivatized, then measured by liquid chromatography (LC)/ESI-MS/MS with selected reaction monitoring.

Results

The derivatized 3α,5α-Adiol, i.e., the bis[(4-dimethylamino)phenyl carbamate] derivative, provided the intense doubly-protonated molecule as the precursor ion, then the specific product ion containing the 3α,5α-Adiol-skeleton by collision-induced dissociation. The detectability of 3α,5α-Adiol was eventually increased 1000-fold by derivatization. Separation of the derivatized 3α,5α-Adiol from its stereoisomers and interfering brain components was achieved using a SunShell Biphenyl column with an isopropyl alcohol-containing mobile phase. A good linearity in the sufficient concentration range, acceptable precision and accuracy, and negligible matrix effect were demonstrated by the validation tests. The animal (rat) study using this method revealed that the brain 3α,5α-Adiol levels were unaffected by the administration of fluoxetine (FLX) and clozapine (CLZ), in contrast to the significant increase of the AP levels.

Conclusion

An LC/ESI-MS/MS method capable of quantifying 3α,5α-Adiol in the rat brain using a 20-mg tissue was developed and validated. The brain levels of 3α,5α-Adiol had an entirely different behavior from those of AP due to FLX and CLZ administration.

理由：5α-雄甾烷-3α,17β-二醇（3α,5α-Adiol）是一种由睾酮衍生的神经类固醇，与由孕酮衍生的神经类固醇异丙孕酮（AP）一样，通过γ-氨基丁酸 A 型受体具有抗焦虑和镇痛作用。虽然人们对精神药物引起的大脑 AP 浓度变化进行了深入研究，但对大脑中 3α,5α-二醇浓度的变化仍然知之甚少。造成这种情况的原因之一是，目前用于定量检测脑内 3α,5α-Adiol、具有足够灵敏度和特异性的有效方法非常有限：为了提高电喷雾离子化-串联质谱法（ESI-MS/MS）对3α,5α-蝶醇的检测能力，采用了4-二甲氨基苯甲酰基叠氮化物进行衍生化。脑样品经固相萃取净化后，对回收的 3α,5α-蝶醇和氘代内标物进行衍生化，然后用液相色谱/ESI-MS/MS 进行选择反应监测：结果：衍生化的 3α,5α-Adiol（即双[(4-二甲基氨基)苯基氨基甲酸酯]衍生物）提供了强烈的双质子化分子作为前体离子，然后通过碰撞诱导解离得到了含有 3α,5α-Adiol-骨架的特定产物离子。通过衍生化，3α,5α-Adiol 的可检测性最终提高了 1000 倍。衍生化后的 3α,5α-二醇与其立体异构体和干扰脑成分的分离是使用 SunShell 联苯色谱柱和含异丙醇的流动相实现的。验证测试表明，该方法在足够的浓度范围内线性关系良好，精密度和准确度均可接受，基质效应可忽略不计。使用该方法进行的动物（大鼠）研究表明，服用氟西汀（FLX）和氯氮平（CLZ）不会影响大脑中 3α,5α-甲二醇的含量，而 AP 的含量则会显著增加：结论：本研究开发并验证了一种能够利用20毫克组织对大鼠大脑中的3α,5α-二醇进行定量的LC/ESI-MS/MS方法。大鼠大脑中的 3α,5α-二醇含量与服用 FLX 和 CLZ 后的 AP 含量完全不同。

{"title":"Conversion of 5α-Androstane-3α,17β-diol to bis[(4-dimethylamino)phenyl carbamate] derivative for sensitive determination of its rat brain level by LC/ESI-MS/MS","authors":"Tatsuya Higashi, Asuka Tanaka, Shiho Tsubura, Shoichi Nishimoto-Kusunose","doi":"10.1002/rcm.9875","DOIUrl":"10.1002/rcm.9875","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>5α-Androstane-3α,17β-diol (3α,5α-Adiol) is a testosterone-derived neurosteroid and has anxiolytic and analgesic effects via γ-aminobutyric acid type A receptors as with the progesterone-derived neurosteroid, allopregnanolone (AP). Although the psychotropic drug-evoked changes in the brain AP concentration have been intensively studied, those in the brain 3α,5α-Adiol concentration remain poorly understood. One of the causes for this is the limited availability of a validated method for quantifying the brain 3α,5α-Adiol with a sufficient sensitivity and specificity, which is described in this study.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>To enhance the detectability of 3α,5α-Adiol by electrospray ionization-tandem mass spectrometry (ESI-MS/MS), derivatization with 4-dimethylaminobenzoyl azide was employed. The brain sample was purified by solid-phase extraction and the recovered 3α,5α-Adiol and the deuterated internal standard were derivatized, then measured by liquid chromatography (LC)/ESI-MS/MS with selected reaction monitoring.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The derivatized 3α,5α-Adiol, i.e., the bis[(4-dimethylamino)phenyl carbamate] derivative, provided the intense doubly-protonated molecule as the precursor ion, then the specific product ion containing the 3α,5α-Adiol-skeleton by collision-induced dissociation. The detectability of 3α,5α-Adiol was eventually increased 1000-fold by derivatization. Separation of the derivatized 3α,5α-Adiol from its stereoisomers and interfering brain components was achieved using a SunShell Biphenyl column with an isopropyl alcohol-containing mobile phase. A good linearity in the sufficient concentration range, acceptable precision and accuracy, and negligible matrix effect were demonstrated by the validation tests. The animal (rat) study using this method revealed that the brain 3α,5α-Adiol levels were unaffected by the administration of fluoxetine (FLX) and clozapine (CLZ), in contrast to the significant increase of the AP levels.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>An LC/ESI-MS/MS method capable of quantifying 3α,5α-Adiol in the rat brain using a 20-mg tissue was developed and validated. The brain levels of 3α,5α-Adiol had an entirely different behavior from those of AP due to FLX and CLZ administration.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756059","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Sulfur isotope analyses using 3× elemental analysis/isotope ratio mass spectrometry: Saving helium and energy while reducing analytical time and costs 利用 3× 元素分析/同位素比质谱法进行硫同位素分析：节省氦气和能源，同时减少分析时间和成本。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-23 DOI: 10.1002/rcm.9866

Jorge E. Spangenberg, Alice Bosco-Santos

Rationale

Helium (He) and energy shortages have caused price increases and reduced their availability. Using three combustion reactions per acquisition of carbon and nitrogen isotope ratios saves 50% He and energy during the elemental analysis/isotope ratio mass spectrometry (EA/IRMS). This approach needs to be tested for sulfur isotope (δ³⁴S) analyses.

Methods

A new method to measure δ³⁴S in three sequential combustion reactions within one EA/IRMS acquisition was developed. The same material or blank samples could be used in the three reactions. After SO₂ was used, a N₂ purging method was employed to prolong the lifetime of the valves in the EA/IRMS interface. The 3×EA/IRMS was applied to measure δ³⁴S in precious samples, such as Ag₂S from acid-volatile and chromium-reducible sulfur extracted with a multiple-port setup.

Results

The 3×EA/IRMS-δ³⁴S method was validated with replicate analyses of international reference materials and laboratory standards with a wide range of mineralogical compositions and δ³⁴S values. The method provided a strategic advantage for the δ³⁴S measurements of small precious samples (measured between blanks). The accuracy and precision of the 3×EA/IRMS values effectively matched those obtained using conventional EA/IRMS, with good agreement between the mean ± SD values and the recommended values with their uncertainties.

Conclusions

Compared with the conventional EA/IRMS, the proposed method provides accurate and precise δ³⁴S measurements of the sulfate and sulfide samples while saving approximately 50% of He, energy, SO₂ reference gas, O₂, analysis time, and cost. Notably, 3×EA/IRMS can provide up to three δ³⁴S values unaffected by memory effects.

理由：氦气（He）和能源短缺导致价格上涨，减少了氦气的供应。在进行元素分析/同位素比质谱分析（EA/IRMS）时，每次采集碳和氮同位素比使用三次燃烧反应可节省 50%的氦气和能源。这种方法需要在硫同位素（δ34S）分析中进行测试：方法：开发了一种在一次 EA/IRMS 采集中测量三个连续燃烧反应中 δ34S 的新方法。三个反应可使用相同的材料或空白样品。使用 SO2 后，采用 N2 吹扫法延长 EA/IRMS 接口阀门的使用寿命。3×EA/IRMS 被用于测量贵重样品中的δ34S，如用多端口装置从酸挥发性和铬还原性硫中提取的 Ag2S：结果：3×EA/IRMS-δ34S 方法通过对具有多种矿物成分和δ34S 值的国际参考材料和实验室标准物质的重复分析进行了验证。该方法为小型贵重样品的 δ34S 测量（在空白之间测量）提供了战略优势。3×EA/IRMS 值的准确度和精确度与使用传统 EA/IRMS 所获得的值有效匹配，平均值 ± SD 值与推荐值及其不确定度之间具有良好的一致性：结论：与传统的 EA/IRMS 相比，所提出的方法能准确测量硫酸盐和硫化物样品的 δ34S 值，同时节省约 50% 的 He、能源、SO2 基准气体、O2、分析时间和成本。值得注意的是，3×EA/IRMS 可提供多达三个不受记忆效应影响的 δ34S 值。

{"title":"Sulfur isotope analyses using 3× elemental analysis/isotope ratio mass spectrometry: Saving helium and energy while reducing analytical time and costs","authors":"Jorge E. Spangenberg, Alice Bosco-Santos","doi":"10.1002/rcm.9866","DOIUrl":"10.1002/rcm.9866","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>Helium (He) and energy shortages have caused price increases and reduced their availability. Using three combustion reactions per acquisition of carbon and nitrogen isotope ratios saves 50% He and energy during the elemental analysis/isotope ratio mass spectrometry (EA/IRMS). This approach needs to be tested for sulfur isotope (<i>δ</i><sup>34</sup>S) analyses.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>A new method to measure <i>δ</i><sup>34</sup>S in three sequential combustion reactions within one EA/IRMS acquisition was developed. The same material or blank samples could be used in the three reactions. After SO<sub>2</sub> was used, a N<sub>2</sub> purging method was employed to prolong the lifetime of the valves in the EA/IRMS interface. The 3×EA/IRMS was applied to measure <i>δ</i><sup>34</sup>S in precious samples, such as Ag<sub>2</sub>S from acid-volatile and chromium-reducible sulfur extracted with a multiple-port setup.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The 3×EA/IRMS-<i>δ</i><sup>34</sup>S method was validated with replicate analyses of international reference materials and laboratory standards with a wide range of mineralogical compositions and <i>δ</i><sup>34</sup>S values. The method provided a strategic advantage for the <i>δ</i><sup>34</sup>S measurements of small precious samples (measured between blanks). The accuracy and precision of the 3×EA/IRMS values effectively matched those obtained using conventional EA/IRMS, with good agreement between the mean ± SD values and the recommended values with their uncertainties.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Compared with the conventional EA/IRMS, the proposed method provides accurate and precise <i>δ</i><sup>34</sup>S measurements of the sulfate and sulfide samples while saving approximately 50% of He, energy, SO<sub>2</sub> reference gas, O<sub>2</sub>, analysis time, and cost. Notably, 3×EA/IRMS can provide up to three <i>δ</i><sup>34</sup>S values unaffected by memory effects.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/rcm.9866","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141746983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterization of metabolic features and potential anti-osteoporosis mechanism of pinoresinol diglucoside using metabolite profiling and network pharmacology 利用代谢物分析和网络药理学研究松脂醇二葡萄糖苷的代谢特征和潜在抗骨质疏松症机制。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-23 DOI: 10.1002/rcm.9872

Xin-Pu Tu, Si-Xian Wu, Meng-Yin Li, Zi-Hao Chen, Cheng-Jun Liu, Yan-Jie Ruan, Jian-Bin Zeng, Wei Shi, Jian-Hang Liu, Feng-Xiang Zhang

Rationale

Eucommia cortex is the core herb in traditional Chinese medicine preparations for the treatment of osteoporosis. Pinoresinol diglucoside (PDG), the quality control marker and the key pharmacodynamic component in Eucommia cortex, has attracted global attention because of its definite effects on osteoporosis. However, the in vivo metabolic characteristics of PDG and its anti-osteoporotic mechanism are still unclear, restricting its development and application.

Methods

Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to analyze the metabolic characteristics of PDG in rats, and its anti-osteoporosis targets and mechanism were predicted using network pharmacology.

Results

A total of 51 metabolites were identified or tentatively characterized in rats after oral administration of PDG (10 mg/kg/day), including 9 in plasma, 28 in urine, 13 in feces, 10 in liver, 4 in heart, 3 in spleen, 11 in kidneys, and 5 in lungs. Furan-ring opening, dimethoxylation, glucuronidation, and sulfation were the main metabolic characteristics of PDG in vivo. The potential mechanism of PDG against osteoporosis was predicted using network pharmacology. PDG and its metabolites could regulate BCL2, MARK3, ALB, and IL6, involving PI3K-Akt signaling pathway, estrogen signaling pathway, and so on.

Conclusions

This study was the first to demonstrate the metabolic characteristics of PDG in vivo and its potential anti-osteoporosis mechanism, providing the data for further pharmacological validation of PDG in the treatment of osteoporosis.

理论依据：杜仲皮是治疗骨质疏松症的传统中药制剂的核心药材。杜仲二葡萄糖苷（Pinoresinol diglucoside，PDG）是杜仲中的质量控制标志物和关键药效学成分，因其对骨质疏松症有确切疗效而备受全球关注。然而，PDG 的体内代谢特征及其抗骨质疏松机制尚不清楚，限制了其开发和应用：方法：采用超高效液相色谱-四极杆飞行时间质谱法分析 PDG 在大鼠体内的代谢特征，并利用网络药理学预测其抗骨质疏松的靶点和机制：结果：大鼠口服PDG（10 mg/kg/天）后，共鉴定或初步鉴定出51种代谢物，其中血浆9种、尿液28种、粪便13种、肝脏10种、心脏4种、脾脏3种、肾脏11种、肺脏5种。呋喃环开放、二甲氧基化、葡萄糖醛酸化和硫酸化是 PDG 在体内的主要代谢特征。利用网络药理学预测了PDG防治骨质疏松症的潜在机制。PDG及其代谢产物可调控BCL2、MARK3、ALB和IL6，涉及PI3K-Akt信号通路、雌激素信号通路等：该研究首次证明了PDG在体内的代谢特征及其潜在的抗骨质疏松症机制，为进一步验证PDG治疗骨质疏松症的药理作用提供了数据。

{"title":"Characterization of metabolic features and potential anti-osteoporosis mechanism of pinoresinol diglucoside using metabolite profiling and network pharmacology","authors":"Xin-Pu Tu, Si-Xian Wu, Meng-Yin Li, Zi-Hao Chen, Cheng-Jun Liu, Yan-Jie Ruan, Jian-Bin Zeng, Wei Shi, Jian-Hang Liu, Feng-Xiang Zhang","doi":"10.1002/rcm.9872","DOIUrl":"10.1002/rcm.9872","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p><i>Eucommia</i> cortex is the core herb in traditional Chinese medicine preparations for the treatment of osteoporosis. Pinoresinol diglucoside (PDG), the quality control marker and the key pharmacodynamic component in <i>Eucommia</i> cortex, has attracted global attention because of its definite effects on osteoporosis. However, the in vivo metabolic characteristics of PDG and its anti-osteoporotic mechanism are still unclear, restricting its development and application.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to analyze the metabolic characteristics of PDG in rats, and its anti-osteoporosis targets and mechanism were predicted using network pharmacology.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>A total of 51 metabolites were identified or tentatively characterized in rats after oral administration of PDG (10 mg/kg/day), including 9 in plasma, 28 in urine, 13 in feces, 10 in liver, 4 in heart, 3 in spleen, 11 in kidneys, and 5 in lungs. Furan-ring opening, dimethoxylation, glucuronidation, and sulfation were the main metabolic characteristics of PDG in vivo. The potential mechanism of PDG against osteoporosis was predicted using network pharmacology. PDG and its metabolites could regulate BCL2, MARK3, ALB, and IL6, involving PI3K-Akt signaling pathway, estrogen signaling pathway, and so on.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study was the first to demonstrate the metabolic characteristics of PDG in vivo and its potential anti-osteoporosis mechanism, providing the data for further pharmacological validation of PDG in the treatment of osteoporosis.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 19","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141750691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Combined matrix-assisted laser desorption/ionisation-mass spectrometry imaging with liquid chromatography-tandem mass spectrometry for observing spatial distribution of lipids in whole Caenorhabditis elegans 基质辅助激光解吸电离质谱成像与液相色谱-串联质谱联用技术，用于观察整条秀丽隐杆线虫体内脂质的空间分布。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-22 DOI: 10.1002/rcm.9850

Michiel Vandenbosch, Erika R. Amstalden van Hove, Ronny Mohren, Isabeau Vermeulen, Henry Dijkman, Ron M. A. Heeren, Pim E. G. Leonards, Samantha Hughes

Rationale

Matrix-assisted laser desorption/ionisation-mass spectrometry imaging (MALDI-MSI) is a powerful label-free technique for biomolecule detection (e.g., lipids), within tissue sections across various biological species. However, despite its utility in many applications, the nematode Caenorhabditis elegans is not routinely used in combination with MALDI-MSI. The lack of studies exploring spatial distribution of biomolecules in nematodes is likely due to challenges with sample preparation.

Methods

This study developed a sample preparation method for whole intact nematodes, evaluated using cryosectioning of nematodes embedded in a 10% gelatine solution to obtain longitudinal cross sections. The slices were then subjected to MALDI-MSI, using a RapifleX Tissuetyper in positive and negative polarities. Samples were also prepared for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using an Exploris 480 coupled to a HPLC Vanquish system to confirm the MALDI-MSI results.

Results

An optimised embedding method was developed for longitudinal cross-sectioning of individual worms. To obtain longitudinal cross sections, nematodes were frozen at −80°C so that all worms were rod shaped. Then, the samples were defrosted and transferred to a 10% gelatine matrix in a cryomold; the worms aligned, and the whole cryomold submerged in liquid nitrogen. Using MALDI-MSI, we were able to observe the distribution of lipids within C. elegans, with clear differences in their spatial distribution at a resolution of 5 μm. To confirm the lipids from MALDI-MSI, age-matched nematodes were subjected to LC-MS/MS. Here, 520 lipids were identified using LC-MS/MS, indicating overlap with MALDI-MSI data.

Conclusions

This optimised sample preparation technique enabled (un)targeted analysis of spatially distributed lipids within individual nematodes. The possibility to detect other biomolecules using this method thus laid the basis for prospective preclinical and toxicological studies on C. elegans.

原理：基质辅助激光解吸电离质谱成像（MALDI-MSI）是一种强大的无标记技术，可用于检测各种生物物种组织切片中的生物大分子（如脂质）。然而，尽管 MALDI-MSI 在许多应用中都很有用，但线虫却没有被常规地与 MALDI-MSI 结合使用。缺乏对线虫体内生物大分子空间分布的研究可能是由于样品制备方面的挑战：本研究开发了一种完整线虫的样品制备方法，评估方法是将线虫嵌入 10%明胶溶液中冷冻切片，获得纵向横截面。然后使用 RapifleX Tissuetyper 在正负极性下对切片进行 MALDI-MSI。此外，还使用与 HPLC Vanquish 系统相连的 Exploris 480 对样品进行液相色谱-串联质谱（LC-MS/MS）分析，以确认 MALDI-MSI 结果：开发了一种优化的包埋方法，用于单个蠕虫的纵向横截面。为获得纵向横截面，线虫在-80°C下冷冻，使所有蠕虫呈棒状。然后，将样本解冻并转移到低温模中的 10%明胶基质中；将蠕虫对齐，并将整个低温模浸没在液氮中。利用 MALDI-MSI，我们能够观察到虫体内脂质的分布，在 5 μm 的分辨率下，脂质的空间分布存在明显差异。为了确认 MALDI-MSI 中的脂质，我们对年龄匹配的线虫进行了 LC-MS/MS。在这里，利用 LC-MS/MS 鉴定出了 520 种脂质，表明与 MALDI-MSI 数据有重叠：结论：这种优化的样品制备技术能够对线虫个体内部空间分布的脂质进行（非）靶向分析。因此，利用这种方法检测其他生物大分子的可能性为对线虫进行前瞻性临床前研究和毒理学研究奠定了基础。

{"title":"Combined matrix-assisted laser desorption/ionisation-mass spectrometry imaging with liquid chromatography-tandem mass spectrometry for observing spatial distribution of lipids in whole Caenorhabditis elegans","authors":"Michiel Vandenbosch, Erika R. Amstalden van Hove, Ronny Mohren, Isabeau Vermeulen, Henry Dijkman, Ron M. A. Heeren, Pim E. G. Leonards, Samantha Hughes","doi":"10.1002/rcm.9850","DOIUrl":"10.1002/rcm.9850","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>Matrix-assisted laser desorption/ionisation-mass spectrometry imaging (MALDI-MSI) is a powerful label-free technique for biomolecule detection (e.g., lipids), within tissue sections across various biological species. However, despite its utility in many applications, the nematode <i>Caenorhabditis elegans</i> is not routinely used in combination with MALDI-MSI. The lack of studies exploring spatial distribution of biomolecules in nematodes is likely due to challenges with sample preparation.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>This study developed a sample preparation method for whole intact nematodes, evaluated using cryosectioning of nematodes embedded in a 10% gelatine solution to obtain longitudinal cross sections. The slices were then subjected to MALDI-MSI, using a RapifleX Tissuetyper in positive and negative polarities. Samples were also prepared for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using an Exploris 480 coupled to a HPLC Vanquish system to confirm the MALDI-MSI results.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>An optimised embedding method was developed for longitudinal cross-sectioning of individual worms. To obtain longitudinal cross sections, nematodes were frozen at −80°C so that all worms were rod shaped. Then, the samples were defrosted and transferred to a 10% gelatine matrix in a cryomold; the worms aligned, and the whole cryomold submerged in liquid nitrogen. Using MALDI-MSI, we were able to observe the distribution of lipids within <i>C. elegans</i>, with clear differences in their spatial distribution at a resolution of 5 μm. To confirm the lipids from MALDI-MSI, age-matched nematodes were subjected to LC-MS/MS. Here, 520 lipids were identified using LC-MS/MS, indicating overlap with MALDI-MSI data.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This optimised sample preparation technique enabled (un)targeted analysis of spatially distributed lipids within individual nematodes. The possibility to detect other biomolecules using this method thus laid the basis for prospective preclinical and toxicological studies on <i>C. elegans</i>.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 17","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/rcm.9850","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Potency analysis of twelve cannabinoids in industrial hemp via ultrahigh-performance liquid chromatography–tandem mass spectrometry 通过超高效液相色谱-串联质谱法对工业大麻中的十二种大麻素进行效力分析。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-22 DOI: 10.1002/rcm.9871

Youxi Cai, Ruiting Zhang, Hao Zhang, Xiaolei Li

Rationale

With an increasing appreciation for the unique pharmacological properties associated with distinct, individual cannabinoids of Cannabis sativa, there is demand for accurate and reliable quantification for a growing number of them. In this study, we developed rapid, sensitive, selective, accurate, and validated liquid chromatography-tandem mass spectrometry for the quantification of cannabinoids.

Methods

Crushed industrial hemp flower and leaf sample was extracted by 95% methanol aqueous, sonicated for 30 min. UPLC-MS/MS analysis using Waters Acquity BEH-C18 column and electrospray ionization(ESI) mass spectrometry detector.

Results

The method was validated to demonstrate its reproducibility and precision, linearity, recovery investigation, and investigation of matrix effect. The concentration–response relationship for all analyzed cannabinoids were linear with R2 values >0.99, with intra- and inter-day precision and relative errors below 12%. The recovery and matrix effect were measured as 66.1%–104.1% and 70.42%–110.75%.

Conclusions

This study established a UHPLC-MS/MS method for the simultaneous and rapid quantitative determination of twelve cannabinoids in industrial hemp flowers and leaves in 11 min. The method was used to analyze 43 industrial hemp flower and leaf samples, with the data being statistically analyzed. Based on the statistical analysis of the cannabinoids, hemp from different regions and different varieties were well distinguished by the PLS-DA model, with the main contributing substances being cannabidiol, Δ9-tetrahydrocannabinol, and Δ8-tetrahydrocannabinol.

理由：随着人们越来越认识到大麻中不同的单个大麻素所具有的独特药理特性，对越来越多的大麻素进行准确可靠定量的需求也在不断增加。在这项研究中，我们开发了快速、灵敏、选择性强、准确且经过验证的液相色谱-串联质谱法，用于定量分析大麻素：方法：用 95% 的甲醇水溶液提取粉碎的工业大麻花和大麻叶样品，超声处理 30 分钟。采用 Waters Acquity BEH-C18 色谱柱和电喷雾离子化（ESI）质谱检测器进行 UPLC-MS/MS 分析：验证了该方法的重现性和精密度、线性关系、回收率调查和基质效应调查。所有分析的大麻素的浓度-反应关系均呈线性，R2 值大于 0.99，日内和日间精密度和相对误差低于 12%。测定的回收率和基质效应分别为 66.1%-104.1%和 70.42%-110.75%：本研究建立了一种超高效液相色谱-质谱/质谱法，可在 11 分钟内同时快速定量测定工业大麻花和叶中的 12 种大麻素。使用该方法分析了 43 份工业大麻花和大麻叶样品，并对数据进行了统计分析。根据对大麻素的统计分析，不同地区和不同品种的大麻通过 PLS-DA 模型得到了很好的区分，主要贡献物质为大麻二酚、Δ9-四氢大麻酚和Δ8-四氢大麻酚。

{"title":"Potency analysis of twelve cannabinoids in industrial hemp via ultrahigh-performance liquid chromatography–tandem mass spectrometry","authors":"Youxi Cai, Ruiting Zhang, Hao Zhang, Xiaolei Li","doi":"10.1002/rcm.9871","DOIUrl":"10.1002/rcm.9871","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Rationale</h3>\u0000 \u0000 <p>With an increasing appreciation for the unique pharmacological properties associated with distinct, individual cannabinoids of Cannabis sativa, there is demand for accurate and reliable quantification for a growing number of them. In this study, we developed rapid, sensitive, selective, accurate, and validated liquid chromatography-tandem mass spectrometry for the quantification of cannabinoids.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Crushed industrial hemp flower and leaf sample was extracted by 95% methanol aqueous, sonicated for 30 min. UPLC-MS/MS analysis using Waters Acquity BEH-C18 column and electrospray ionization(ESI) mass spectrometry detector.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The method was validated to demonstrate its reproducibility and precision, linearity, recovery investigation, and investigation of matrix effect. The concentration–response relationship for all analyzed cannabinoids were linear with R2 values >0.99, with intra- and inter-day precision and relative errors below 12%. The recovery and matrix effect were measured as 66.1%–104.1% and 70.42%–110.75%.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study established a UHPLC-MS/MS method for the simultaneous and rapid quantitative determination of twelve cannabinoids in industrial hemp flowers and leaves in 11 min. The method was used to analyze 43 industrial hemp flower and leaf samples, with the data being statistically analyzed. Based on the statistical analysis of the cannabinoids, hemp from different regions and different varieties were well distinguished by the PLS-DA model, with the main contributing substances being cannabidiol, Δ9-tetrahydrocannabinol, and Δ8-tetrahydrocannabinol.</p>\u0000 </section>\u0000 </div>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":"38 18","pages":""},"PeriodicalIF":1.8,"publicationDate":"2024-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tracing the geographic origin of Atlantic cod products using stable isotope analysis. 利用稳定同位素分析追踪大西洋鳕鱼产品的地理来源。

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry

Pub Date : 2024-07-22 DOI: 10.1002/rcm.9861

Juliet S E Wilson, Rona A R McGill, Petur Steingrund, Clive N Trueman

Rationale: Increasing demand for fish and seafood means that the traceability of marine products is becoming ever more important for consumers, producers and regulators. Highly complex and globalised supply networks create challenges for verifying a stated catch region. Atlantic cod is one of the most commercially important species in the northeast Atlantic. Several regional fisheries supply cod into the trade network, of which some are at greater risk of overexploitation than others. Tools allowing retrospective testing of spatial origin would significantly assist sustainable harvesting of fish, reducing incentives for illegal fishing and fraud.

Methods: Here, we investigate whether stable isotope ratios of carbon, nitrogen and sulphur can be used to retrospectively identify the catch region of Atlantic cod (Gadus morhua). We measured the isotopic composition of muscle tissue from 377 cod from 10 catch regions across the northeast Atlantic and then applied three different assignment methods to classify cod by region of most likely origin. The assignment method developed was subsequently tested using independently sourced, known-origin samples.

Results: Individual cod could be traced back to their true origin with an average assignment accuracy of 70-79% and over 90% accuracy for certain regions. Assignment success rates comparable to those using genetic techniques were achieved when assigning among restricted and pre-selected regions. However, assignment accuracy to the fishery region estimated from independent samples across the whole geographic range of cod averaged ~25% overall, highlighting the need for careful application of isotope-based approaches.

Conclusion: Stable isotope techniques can provide effective tools to test for origin in Atlantic cod, but not all catch regions are isotopically distinct. Stable isotopes could be combined with genetic techniques to result in higher assignment accuracy than could be achieved using either method independently. Assignment potential can be estimated from reference datasets, but estimates of realistic assignment accuracy require independently collected data.

理由：对鱼类和海产品日益增长的需求意味着，海产品的可追溯性对消费者、生产者和监管者来说变得越来越重要。高度复杂和全球化的供应网络给验证既定捕捞区域带来了挑战。大西洋鳕鱼是大西洋东北部最重要的商业鱼种之一。多个区域渔场向贸易网络供应鳕鱼，其中一些渔场比其他渔场面临更大的过度开发风险。方法：在此，我们研究了碳、氮和硫的稳定同位素比值是否可用于回顾性地确定大西洋鳕鱼（Gadus morhua）的捕捞区域。我们测量了来自大西洋东北部 10 个捕获区的 377 条鳕鱼肌肉组织的同位素组成，然后采用三种不同的分配方法按鳕鱼最有可能的来源区进行分类。随后，我们使用独立来源的已知原产地样本对所开发的分配方法进行了测试：结果：鳕鱼个体可追溯到其真正的原产地，平均准确率为 70-79%，某些地区的准确率超过 90%。在限制区域和预选区域之间进行分配时，分配成功率可与使用基因技术的分配成功率相媲美。然而，在整个鳕鱼地理范围内，根据独立样本估计的渔业区域分配准确率总体平均约为 25%，这突出表明需要谨慎应用基于同位素的方法：结论：稳定同位素技术可为检测大西洋鳕鱼的产地提供有效工具，但并非所有渔获区域在同位素上都是不同的。稳定同位素可与遗传技术相结合，从而获得比单独使用其中一种方法更高的定位精度。可以根据参考数据集估算分配潜力，但实际分配准确性的估算需要独立收集的数据。

{"title":"Tracing the geographic origin of Atlantic cod products using stable isotope analysis.","authors":"Juliet S E Wilson, Rona A R McGill, Petur Steingrund, Clive N Trueman","doi":"10.1002/rcm.9861","DOIUrl":"https://doi.org/10.1002/rcm.9861","url":null,"abstract":"<p><strong>Rationale: </strong>Increasing demand for fish and seafood means that the traceability of marine products is becoming ever more important for consumers, producers and regulators. Highly complex and globalised supply networks create challenges for verifying a stated catch region. Atlantic cod is one of the most commercially important species in the northeast Atlantic. Several regional fisheries supply cod into the trade network, of which some are at greater risk of overexploitation than others. Tools allowing retrospective testing of spatial origin would significantly assist sustainable harvesting of fish, reducing incentives for illegal fishing and fraud.</p><p><strong>Methods: </strong>Here, we investigate whether stable isotope ratios of carbon, nitrogen and sulphur can be used to retrospectively identify the catch region of Atlantic cod (Gadus morhua). We measured the isotopic composition of muscle tissue from 377 cod from 10 catch regions across the northeast Atlantic and then applied three different assignment methods to classify cod by region of most likely origin. The assignment method developed was subsequently tested using independently sourced, known-origin samples.</p><p><strong>Results: </strong>Individual cod could be traced back to their true origin with an average assignment accuracy of 70-79% and over 90% accuracy for certain regions. Assignment success rates comparable to those using genetic techniques were achieved when assigning among restricted and pre-selected regions. However, assignment accuracy to the fishery region estimated from independent samples across the whole geographic range of cod averaged ~25% overall, highlighting the need for careful application of isotope-based approaches.</p><p><strong>Conclusion: </strong>Stable isotope techniques can provide effective tools to test for origin in Atlantic cod, but not all catch regions are isotopically distinct. Stable isotopes could be combined with genetic techniques to result in higher assignment accuracy than could be achieved using either method independently. Assignment potential can be estimated from reference datasets, but estimates of realistic assignment accuracy require independently collected data.</p>","PeriodicalId":225,"journal":{"name":"Rapid Communications in Mass Spectrometry","volume":" ","pages":"e9861"},"PeriodicalIF":1.8,"publicationDate":"2024-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141746982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0