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Quantification and Carbon and Nitrogen Isotopic Measurements of Heme B in Environmental Samples.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-07-21 , DOI: 10.1021/acs.analchem.0c01711 Yuta Isaji 1 , Nanako O Ogawa 1 , Yoshinori Takano 1 , Naohiko Ohkouchi 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-07-21 , DOI: 10.1021/acs.analchem.0c01711 Yuta Isaji 1 , Nanako O Ogawa 1 , Yoshinori Takano 1 , Naohiko Ohkouchi 1
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Heme B is an iron-coordinated tetrapyrrole molecule that acts as a cofactor in hemoproteins. It is expected to be ubiquitous in the environment, as b-type hemoproteins catalyze a variety of essential biochemical reactions. In this study, we developed an analytical method to quantify heme B in biological and environmental samples using high-performance liquid chromatography (HPLC) coupled to a photodiode array detector. The applicability of our method was further extended by the use of liquid chromatography/mass spectrometry (LC/MS; detection limit: ∼1 fmol), which enabled the quantification of a trace amount of dissolved heme B in filtered seawater and sedimentary heme B coexisting with an abundant interfering organic matrix. For compound-specific carbon and nitrogen isotopic measurements, heme B was successfully isolated and purified from biological and environmental samples by a combination of anion-exchange column chromatography, methyl esterification, and dual-step HPLC. While carbon and nitrogen isotopic compositions of heme B in phototrophs were mostly comparable to those of chlorophyll a, heme B in suspended particulate materials in coastal water and an intertidal sediment was 13C-depleted and 15N-enriched relative to chlorophyll a, suggesting that nonphototrophic microorganisms are also a significant source of heme B in natural environments.
中文翻译：
环境样品中血红素B的定量以及碳和氮同位素测量。
血红素B是铁配位的四吡咯分子，在血红蛋白中起辅助因子的作用。预计到无处不在的环境，为b型血蛋白催化多种基本的生化反应。在这项研究中，我们开发了一种使用高效液相色谱（HPLC）耦合光电二极管阵列检测器来定量分析生物和环境样品中血红素B的分析方法。我们的方法的适用性通过使用液相色谱/质谱法（LC / MS；检测极限：〜1 fmol）进一步扩展，从而能够定量分析过滤后的海水中微量的溶解血红素B和沉积的血红素B共存具有丰富的干扰有机基质。对于化合物特定的碳和氮同位素测量，通过阴离子交换柱色谱，甲基酯化和双步HPLC的结合，成功地从生物和环境样品中分离并纯化了血红素B。a，相对于叶绿素a而言，沿海水和潮间带沉积物中悬浮颗粒物质中的血红素B减少了13 C，N富集了15 N ，这表明非营养型微生物也是自然环境中血红素B的重要来源。
更新日期：2020-08-18
中文翻译：
环境样品中血红素B的定量以及碳和氮同位素测量。
血红素B是铁配位的四吡咯分子，在血红蛋白中起辅助因子的作用。预计到无处不在的环境，为b型血蛋白催化多种基本的生化反应。在这项研究中，我们开发了一种使用高效液相色谱（HPLC）耦合光电二极管阵列检测器来定量分析生物和环境样品中血红素B的分析方法。我们的方法的适用性通过使用液相色谱/质谱法（LC / MS；检测极限：〜1 fmol）进一步扩展，从而能够定量分析过滤后的海水中微量的溶解血红素B和沉积的血红素B共存具有丰富的干扰有机基质。对于化合物特定的碳和氮同位素测量，通过阴离子交换柱色谱，甲基酯化和双步HPLC的结合，成功地从生物和环境样品中分离并纯化了血红素B。a，相对于叶绿素a而言，沿海水和潮间带沉积物中悬浮颗粒物质中的血红素B减少了13 C，N富集了15 N ，这表明非营养型微生物也是自然环境中血红素B的重要来源。
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