Most of analytical community is focused on reversed phase high performance liquid chromatography (RP-HPLC) for mercury speciation by employing mobile phases comprising of high salts and moderate amounts of organic solvents. This study aims at rapid mercury speciation analysis by ion-pairing RP-HPLC with inductively coupled plasma mass spectrometry (ICP-MS) detection only using low salts for the sake of green analytical chemistry. Two ion-pairing HPLC methods were developed on individual usage of positively and negatively charged ion-pairing reagents (tetrabutylammonium hydroxide –TBAH and sodium dodecylbenzene sulfonate –SDBS), where sodium 3-mercapto-1-propysulfonate (MPS) and l-cysteine (Cys) were individually added in mobile phases to transform mercury species into negative and positive Hg-complexes for good resolution. Addition of phenylalanine was also utilized for rapid baseline separation in combination of short C₁₈ guard columns. Optimum mobile phases of 2.0 mM SDBS + 2.0 mM Cys + 1.0 mM Phe (pH 3.0) and 4.0 mM TBAH + 2.0 mM MPS + 2.0 mM Phe (pH 6.0) both achieved baseline separation of inorganic mercury (Hg²⁺), methylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg) on two consecutive 12.5-mm C₁₈ columns. The former mobile phase was selected for mercury speciation in freshwater fish because of short separation time (3.0 min). Detection limits of 0.015 for Hg²⁺, 0.014 for MeHg, 0.028 for EtHg and 0.042 μg L⁻¹ for PhHg were obtained along with satisfactory precisions of peak height and area (1.0-2.8% for 5.0 μg L⁻¹ Hg-mixture standard). Good accordance of determined values of MeHg and total mercury in certified reference materials of fish tissue (GBW 10029) and tuna fish (BCR-463) with certified values as well as good recoveries (91–106%) proved good accuracy of the proposed method. An example application to freshwater fish indicated its potential in routine analysis, where MeHg was presented at 3.7–20.3 μg kg⁻¹ as the dominate species.

通过使用由高盐和适量有机溶剂组成的流动相，大多数分析界将重点放在用于汞形态分析的反相高效液相色谱（RP-HPLC）上。这项研究旨在通过离子对RP-HPLC和电感耦合等离子体质谱（ICP-MS）检测对汞进行快速形态分析，仅出于绿色分析化学的目的而仅使用低盐。针对分别使用带正电和带负电的离子对试剂（氢氧化四丁铵– TBAH和十二烷基苯磺酸钠– SDBS），开发了两种离子对HPLC方法，其中3-巯基-1-丙磺酸钠（MPS）和l在流动相中分别添加-半胱氨酸（Cys），以将汞物质转化为负和正Hg络合物，以获得良好的分离度。在短C ₁₈保护柱的组合中，苯丙氨酸的添加也可用于快速基线分离。最佳流动相2.0 mM SDBS + 2.0 mM Cys + 1.0 mM Phe（pH 3.0）和4.0 mM TBAH + 2.0 mM MPS + 2.0 mM Phe（pH 6.0）均实现了无机汞（Hg ²⁺），甲基汞（ MeHg），乙基汞（EtHg）和苯基汞（PhHg）在两个连续的12.5 mm C ₁₈色谱柱上。由于分离时间短（3.0分钟），因此选择前一种流动相进行淡水鱼中的汞形态分析。Hg ^2+的检出限为0.015获得的MeHg为0.014，EtHg为0.028，PhHg为0.042μgL ^-1，并且峰高和峰面积的精度令人满意（5.0μgL ^-1 Hg混合物标准品的1.0-2.8％）。鱼类组织（GBW 10029）和金枪鱼（BCR-463）的认证参考物质中MeHg和总汞的测定值与认证值的良好符合以及良好的回收率（91–106％）证明了该方法的准确性。淡水鱼的一个实例应用表明了其在常规分析中的潜力，其中甲基汞的主要种类为3.7–20.3μgkg ^-1。