Interfacing nanoliter liquid chromatography and inductively coupled plasma mass spectrometry with an in-column...

Metal speciation analysis of microsamples in the clinical, biological and forensic fields is important for elucidating the metals' toxicity, mobility and metabolic behaviors of metals. Such analysis may be achieved by nanoliter high-performance liquid chromatography (nanoHPLC) hyphenated with inductively coupled plasma mass spectrometry (ICP-MS). In this work, an in-column high-pressure nebulizer (ICHPN) was developed to enhance the sensitivity and the separation efficiency of the sheathless nanoHPLC-ICP-MS coupling. The ICHPN consists of two concentric fused-silica capillaries with tapered tips connected via a PEEK tee, where C₁₈ silica particles with a size of 5 μm were packed in the tapered inner capillary based on the keystone effect. Combining a heated single pass spray chamber with a makeup gas, the ICHPN was capable of independently optimizing the nebulization efficiency and the transport efficiency. The ICHPN offered high sensitivities, low detection limits and good precision at nanoflow rates. Compared with commercial nebulizers, the ICHPN fabrication was simple, rapid, reproducible and inexpensive. By implementing the ICHPN, we achieved rapid separation of four mercury species, i.e., Hg²⁺ and methyl-, ethyl- and phenylmercuric chloride, within 8.0 min with good resolution (2.0-13.9). Detection limits of 0.044-0.13 μg L^-1 were obtained with precisions of peak heights and areas ranging from 1.5 to 3.5% for a 50 μg L^-1 standard solution. Good agreement between the determined and certified values of mercury species in a certified reference material of caprine blood (SRM 955c) together with good recoveries (93-101%) validated the accuracy of the method.