Compact photometric detector integrated with separation microchip for potential portable liquid chromatography system.

In recent years, miniaturized analytical instruments have been developing to meet the needs of portable and rapid analysis. The key of miniaturized analytical equipment is the miniaturization and integration of functional modules. This paper aims to develop a miniaturized photometric detector and separation microfluidic chip for a liquid chromatography (LC) system. The detector uses a light-emitting diode to emit ultraviolet light, which is collimated by an internal double lens. A Z-shaped flow cell with a long optical path is designed and fabricated in the separation microfluidic chip with a three-layer structure, which provides a tubing-free connection between the separation and detection unit. Detector performance is evaluated using hemoglobin (Hb) samples, with an upper limit of detection linearity (95 %) of 0.345 AU and stray light level as low as 0.08 %. Additionally, the microchip channel can be filled with cation exchange resin and C18 particles. Finally, an ion LC system and a reversed-phase LC system were constructed based on the miniaturized photometric detector and two microchips with different packed columns, respectively, and were successfully used in the separation and detection of two metabolic markers (glycated hemoglobin or bilirubin). The results of this study are expected to facilitate the development of a portable LC system and their application in community health services and family health management of chronic diseases.

Rapid detection of glycosylated hemoglobin levels by a microchip liquid chromatography system in gradient elution mode.

BACKGROUND: The determination of glycosylated hemoglobin (HbA1c) is crucial for diabetes diagnosis and can provide more substantial results than the simple measurement of glycemia. While there is a lack of simple methods for the determination of HbA1c using a point-of-care test (POCT) compared to glycemia measurement. In particular, high-performance liquid chromatography (HPLC) is considered the current gold standard for determining HbA1c levels. However, commercial HPLC systems usually have some sort of disadvantages such as bulky size, high-cost and need for qualified operators. Therefore, there is an urgent demand to develop a portable, and fast HbA1c detection system consuming fewer reagents. RESULTS: We present a novel microchip that integrates a micromixer, passive injector, packed column and detection cell. The integrated microchip, in which all the microstructures were formed in the CNC machining center through micro-milling, is small in size (30 mm × 70 mm × 10 mm), and can withstand 1600 psi of liquid pressure. The integrated design is beneficial to reduce the band broadening caused by dead volume. Based on the microchip, a microchip liquid chromatography (LC) system was built and applied to the analysis of HbA1c. The separation conditions of HbA1c in blood calibrator samples were optimized using the microchip LC system. Samples containing four levels of HbA1c were completely separated within 2 min in optimal gradient conditions, with an inaccuracy (<3.2 %), a coefficient of variation (c.v. < 2.1 %) and a correlation coefficient (R2 = 0.993), indicating excellent separation efficiency and reproducibility. SIGNIFICANCE: The POCT of HbA1c is critical for diabetes diagnosis. The microchip chromatography system was developed for HbA1c determination, which contains an integrated microchip and works under a gradient elution. It surpasses existing chip technology in terms of separation performance and detection speed, providing a competitive advantage for POCT of HbA1c. It is considered one important step for realizing efficient portable systems for timely and accurate diabetes diagnosis.