practical problem solving in hplc pdf

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Back to School—Solving Practical HPLC Problems with Basic Theory

I’m very much a “big picture” type of thinker. By that, I mean unless I can understand all of the working parts of a problem and understand how they interact, I find it difficult to decide the best approach to figuring out how to solve the issue. While this is certainly a personality trait, I often wonder how chromatographers with some thirty-plus years less experience than myself begin to fathom the complexities of separation science and so begin to develop their own approach to practical problem solving.

The Purnell Equation shown in Figure 1 does an excellent job of describing the change in chromatographic Resolution ( R ) obtained when altering each of the principle chromatographic variables, namely efficiency ( N ), selectivity ( a ) or retention factor ( k ). When I’m teaching chromatography, it’s at the point we begin to roll out the governing equations that I see that glazed look come over many students, as the old guy “tries to tie everything back to theory,” “when was the last time I ever used an equation in the laboratory,” and “when was the last time he ever did any real chromatography.” Most times I can retrieve this situation by answering, “yesterday,” and by relating the mathematical relationships to the last time they stood in front of their HPLC equipment scratching their head for inspiration! Let’s do the same thing here and see how this particular equation can help with practical problems.

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High-Performance Liquid Chromatography (HPLC)

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Sample Preparation

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Troubleshooting

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Meyer, V.R. (2005). High-Performance Liquid Chromatography (HPLC). In: Dhein, S., Mohr, F.W., Delmar, M. (eds) Practical Methods in Cardiovascular Research. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-26574-0_35

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Review on Common Observed HPLC Troubleshooting Problems

2020, International Journal of Pharma Research and Health Sciences

HPLC provide much higher resolution, more accurate quantitative results, as well as shorter analysis times in comparison to the earlier techniques, HPLC has evolved into an indispensable tool in many analytical laboratories. Troubleshooting is a form of problem solving, often applied to repair failed products or processes.Actually, HPLC refers to a number of separation techniques that use a liquid mobile phase, or eluent. Troubleshooting HPLC instrumentation and separations require a fundamental understanding of how the instrument functions and how the separation works. Common HPLC problems are caused by component malfunctions (pump, degasser, injector, detector, data system, column), and faulty preparation of the mobile phase or sample preparation. Always use well cleaned glassware for preparation for better results. Main key to resolve HPLC problem is Cleaning of HPLC with appropriate solvent. If any problem occurs, it is advisable to perform a quick visual check of the instrument and column. If Monograph or any protocol is provided thentry to follow same process. Troubleshooting is a form of problem solving, often applied to repair failed products or processes. Otherwise, we suggest that you read the entire article so as to pick up some ideas that will help you avoid problems in the future. We hope that this article useful to diagnose problems and to gain understanding of underlying causes so that you can prevent or minimize future occurrence. Colum and HPLC care is most important part of troubleshooting. In This article, we tried to cover all major troubleshooting problems.

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Mohammed Tayeb

Glufosinate ammonium or ammonium salt (ammonium-(2RS)-2-amino-4-(methylphosphinato) butyric acid; C 5 H 15 N 2 O 4 P) is a commonly used polar herbicide in Malaysia and present in a variety of environmental waters at the sub-ppb level. Thus, glufosinate ammonium is analyzed in soil and water using high-performance liquid chromatography (HPLC), which is a complex yet the most powerful analysis tool. HPLC is tremendously sensitive and highly automated and HPLC instrumentation and machinery have improved over the years. However, typical problems are still encountered. HPLC users and advanced learners require help in identifying, separating and correcting typical problems. All HPLC systems consist of similar basic components. Although it is a modular system, trouble can occur in each component and change the overall performance. Resolving these problems may be expensive. This review describes the different aspects of HPLC, particularly troubleshooting, common problems and easy guidelines for maintenance. ABSTRAK Ammonium glufosinate atau garam ammonium (ammonium-(2RS)-2-amino-4-(methylphosphinato) asid butirik; C 5 H 15 N 2 O 4 P) adalah herbisid berkutub yang biasa digunakan di Malaysia dan hadir di dalam persekitaran air pada kepekatan sub-ppb. Oleh itu, ammonium glufosinate dianalisis di dalam tanah dan air dengan menggunakan kromatografi cecair berprestasi tinggi (HPLC) yang merupakan alat yang kompleks namun sangat jitu dalam menganalisis herbisid ini. HPLC sangat sensitif dan automatik serta prestasinya sering dipertingkatkan dari semasa ke semasa. Walau bagaimanapun, terdapat beberapa masalah sering dihadapi pengguna. Pengguna dan pengendali HPLC pada peringkat lebih tinggi memerlukan bantuan dalam proses pengenalpastian, pengasingan dan membetulkan permasalahan yang sering berlaku. Kesemua sistem HPLC terdiri daripada komponen asas yang sama. Walaupun ia adalah satu sistem yang modular, masalah boleh berlaku dalam setiap komponen dan mengubah keseluruhan prestasi. Menyelesaikan masalah ini mungkin memerlukan kos yang tinggi. Maka, ulasan ini menerangkan pelbagai aspek HPLC, terutamanya penyelesaian masalah HPLC, masalah yang sering berlaku dan garis panduan yang mudah untuk penyelenggaraan.

KNonStop Channel

Analyze • Detect • Measure • Control ™ c h r o m a t o g r a p h y

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PHARMANEST JOURNAL

Ahmed Hassan

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Ms. Khuspreet Kaur

zahra hashemi

liliana andrea

Untargeted metabolomics aims to gather information on as many metabolites as possible in biological systems by taking into account all information present in the data sets. Here we describe a detailed protocol for large-scale untargeted metabolomics of plant tissues, based on reversed phase liquid chromatography coupled to high-resolution mass spectrometry (LC-QTOF MS) of aqueous methanol extracts. Dedicated software, MetAlign, is used for automated baseline correction and alignment of all extracted mass peaks across all samples, producing detailed information on the relative abundance of thousands of mass signals representing hundreds of metabolites. Subsequent statistics and bioinformatics tools can be used to provide a detailed view on the differences and similarities between (groups of) samples or to link metabolomics data to other systems biology information, genetic markers and/or specific quality parameters. The complete procedure from metabolite extraction to assembly of a data matrix with aligned mass signal intensities takes about 6 days for 50 samples.

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