A rational approach for HPLC calibration using mass spectrometer as detector

Paresh Varshney , Atul Kaushik , Alok Sharma , Sajal Srivastava , Parmesh Kumar Dwivedi
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Abstract

Background/Objective

A direct, accurate, precise, reproducible and cost-time efficient approach for calibration of HPLC using mass spectrometer as detector has been described which simulates the routine analysis in bioanalytical laboratories.

Methods

Injector position check and injection volume repeatability test for the calibration of autosampler; and pump flow rate accuracy test for the calibration of pumps were illustrated using direct weighing method. Carryover test, pump repeatability test, linearity and precision were carried out for holistic (overall system) calibration of the instrument using a sensitive high throughput LC–MS/MS pre-validated method.

Results/Conclusion

The results of calibration experiments for individual modules suggested that pumps are yielding accurate flow rate, autosampler is aspirating the correct injection volume irrespective to position or volume, and column oven and autosampler chambers are maintaining steady temperatures. Holistic calibration verified the accuracy, precision, repeatability and reproducibility of the instrument with insignificant carryover.

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一种以质谱仪为检测器的高效液相色谱定标方法
背景/目的建立了一种直接、准确、精密度高、重现性好、成本-时间效益高的以质谱仪为检测器的HPLC校准方法,模拟了生物分析实验室的常规分析。方法采用进样器位置检查和进样量重复性试验对自动进样器进行标定;并举例说明了用直接称量法标定泵流量精度的方法。使用灵敏的高通量LC-MS /MS预验证方法对仪器进行了整体(整体系统)校准,进行了结转测试、泵重复性测试、线性和精度测试。结果/结论各模块的标定实验结果表明,泵的流量准确,自动进样器无论位置或体积都能正确吸入进样量,柱箱和自动进样室温度稳定。整体校准验证了仪器的准确性、精密度、重复性和再现性,无显著的结转。
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