Derivatizing amino acids doesn’t have to take hours to do. A more hands-off solution can save your lab hours of prep work. And help you trust your data even more.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_f41f58db04.jpeg" alt="Thermo Fisher Scientific: How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_f41f58db04.jpeg 245w," sizes="100vw" width="1024" height="512">Ever wonder what makes a simple molecule like valine so complicated to measure?Amino acids—the building blocks of proteins—are everywhere. In your food. Body. And even the fertilizers that help grow our food. But measuring these compounds precisely? That’s a classic analytical headache, even for an experienced chromatographer.Ask any lab scientist who’s done amino acid analysis, and they’ll tell you the chromatographic separation isn’t the hard part. The challenge is sample preparation, or everything before the chromatography. Handling toxic reagents. The repeated pipetting. And then timing the derivatization reactions just right.All these manual steps can contribute to lower method precision and data reproducibility.So, what’s the solution? Using a more hands-off approach. You get the same trusted chemical reaction, but your method is now faster, cleaner, and more reliable.Let’s walk through a real-life example of how your lab can derivatize amino acids directly inside the HPLC autosampler needle.<h3>Key takeaways</h3><div><ul><li>Manual derivatization of amino acids is laborious, error-prone, and handling toxic chemicals can pose health hazards to lab staff.</li><li>Automated in-needle OPA/FMOC derivatization of amino acids is a time- and labor-saving approach for HPLC-FLD analysis.</li><li>Reliable method performance, column stability over several hundred injections, and using inexpensive reagents facilitates high sample throughput at low costs per sample for routine laboratories.</li><li>Sample matrices of medium and high complexity (beverages, food, fertilizers) using in-needle derivatization is robust and reproducible.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_2_a906250f79.jpeg" alt="Thermo Fisher Scientific: How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids: Illustration showing manual derivatization versus in-needle derivatization." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_2_a906250f79.jpeg 245w," sizes="100vw" width="700" height="410"><h3>Why does the derivatisation of amino acids still matter when aerosol detectors are an option? The unmatched sensitivity of fluorescence detection compared to evaporative technology.</h3>Most amino acids don’t have double bonds that naturally absorb UV light, meaning your HPLC detector can’t “see” the compounds directly.To solve these visibility issues, chemists use derivatization. The conjugation reaction is brilliant chemistry, facilitating the formation of <a target="_blank" rel="noopener noreferrer" href="https://www.thermofisher.com/content/lifetech/global/en/home/industrial/chromatography/chromatography-learning-center/liquid-chromatography-information/hplc-system-components/how-hplc-detectors-work.html">fluorescent derivatives</a> suitable for sensitive and precise analysis.This technique involves reacting amino acids in the solution for analysis with reagents that make the amino acids fluorescent. The most common pair are OPA (ortho-phthaldialdehyde) for primary amino acids and FMOC-Cl (9-fluorenylmethyloxycarbonyl chloride) for secondary ones.Now here’s what can go wrong. You measure reagents, mix samples by hand, wait for the reaction, then inject onto your HPLC instrument. Small inconsistencies, such as a few seconds too long or a reagent variation, can mean BIG differences in peak response, method accuracy, and data reproducibility. Removing these inconsistencies is where automated in-needle derivatization comes in.<h3>Let your HPLC autosampler needle handle the not-so-friendly laborious tasks</h3>Switching from manual to automated in-needle derivatization can help make this labor-intensive process easier for everyone in your lab, while improving data consistency and overall lab safety. &nbsp;Here’s how:<div><div><ul><li>Instead of manually preparing reaction mixtures, you can use a compatible instrument, like<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=37&amp;manufacturerItems=3&amp;search=vanquish"> Thermo Scientific Vanquish LC Systems</a>, to perform in-needle derivatization.</li><li>And, with the helpful guidance of supporting software, you can automatically mix samples and reagents in the needle, time the reaction more precisely, and inject the derivatized sample onto the column.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_3_5c636b1584.jpeg" alt="Thermo Fisher Scientific: How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids: In-needle derivatization of primary and secondary AAs with OPA/MPA and FMOC-Cl reagents for subsequent LC analysis of the fluorescent derivatives. *Borate buffer pH 10, **acetic acid for quenching." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_3_5c636b1584.jpeg 245w," sizes="100vw" width="768" height="384"><h3>A real-life example showcasing better efficiency, robustness, and throughput capabilities.</h3>What makes this automated workflow stand out isn’t a new reaction—but rather a faster way of working with your HPLC instrument.The reactions between OPA/MPA and FMOC-Cl still happen as before, but now the reaction is controlled digitally through <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/cn_003698_hplc_vanquish_workflow_amino_acid_analysis_derivatization_cn003698_na_en_e76e3abe44.pdf">Thermo Scientific Chromeleon Chromatography Data System</a> (CDS).As demonstrated in our whitepaper, each reagent draw, mix, and injection step is defined in a Custom Injection Program (CIP), eliminating timing errors and variability. <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/76">Chromeleon CDS</a> visualizes every step in its Instrument Method Editor, tracks parameters in real time, and logs events in the Audit Trail for compliance.And since the workflow design has flexibility in mind, you can use Chromeleon CDS or a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ab_004093_hplc_vanquish_core_amino_acids_needle_derivatization_ab004093_na_en_edb37df857.pdf">third-party software</a>, with identical parameters, which gives your labs the freedom to work within your preferred environment without retraining staff or rewriting SOPs.<h3>Trusting your data and a faster derivatization technique. That’s the science behind putting your sample preparation inside the needle.</h3>The most beneficial part of this workflow is the hours of time your lab and broader QC team will save.<ul><li>Consistent fluorescence response and accurate quantitation for all amino acids</li><li>Excellent injection-to-injection reproducibility, even across reagent batches</li><li>Improved safety through reduced manual solvent handling</li><li>Less hands-on time, freeing analysts for higher-value work</li><li>Seamless CDS integration for full audit traceability</li></ul>So next time you’re pipetting derivatization reagents by hand, secretly wishing there was a better way, why not consider an automated method approach?You get the same chemistry, performed automatically inside the autosampler, hands-free, ready to inject in minutes with reproducible data that you can trust.Learn more about this new automated amino acid analytical workflow in our <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/cn_003698_hplc_vanquish_workflow_amino_acid_analysis_derivatization_cn003698_na_en_e76e3abe44.pdf">customer application note today</a>.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_4_341aed0b3a.jpeg" alt="Thermo Fisher Scientific: How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids: Chromatographic results of calibration standard (50 µM) measured with the Vanquish Core HPLC system, comparing method performance on 1st and 14th day of measurements." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_How_to_Cut_HPLC_Sample_Preparation_Time_for_Derivatizing_Amino_Acids_4_341aed0b3a.jpeg 245w," sizes="100vw" width="768" height="384"><h3>Helpful tips to improve method robustness with Vanquish LC Systems+ Chromeleon CDS</h3><ol><li>Use a 100 µL sample loop for thorough and reproducible mixing of the sample and reagents during the derivatization step.</li><li>Install a passive pre-heater to facilitate proper dilution of the reagents before the column.</li><li>Use guard cartridges for your columns and replace the cartridges when column performance diminishes. Apply smart tools like intelligent run control in Chromeleon CDS for automated checks.</li><li>Replace derivatization reagents in your autosampler daily.</li><li>Periodically flush your column with 100% acetonitrile for 30 minutes to remove persistent reaction products and matrices.</li></ol><blockquote>Visit us on&nbsp;<a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/">LinkedIn:</a>&nbsp;#AutomatedSamplePreparation #HPLC</blockquote></div></div></div>

How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids

<h3>How long does it actually take to install all the capillaries and fittings on a conventional HPLC system?</h3><h4>The Small Components That Matter Most</h4>When people think about HPLC systems, they often focus on pumps, detectors, columns, or software. Capillaries and fittings are usually seen as minor components because they are relatively inexpensive and physically small. However, every chromatographer eventually discovers that these tiny parts can have a major influence on system performance.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Connecting_a_U_HPLC_system_can_be_difficult_at_some_poorly_accessible_locations_3b837b441b.webp" alt="KNAUER: Connecting a (U)HPLC system can be difficult at some poorly accessible locations" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Connecting_a_U_HPLC_system_can_be_difficult_at_some_poorly_accessible_locations_3b837b441b.webp 208w," sizes="100vw" width="720" height="540">Even decades ago, chromatographers recognized the importance of proper connections. In the 1980s, long before UHPLC became common, John Dolan wrote about minimizing dead volume through correct fitting installation. Today, with UHPLC systems operating at extremely high pressures and using columns with very small particle sizes, dead volume is even less tolerable than before. The topic remains just as relevant today as it was back then.Interestingly, despite what many users assume, the chromatographic column itself is often not the primary source of HPLC or UHPLC problems. In my experience, poor connections are far more commonly responsible for issues such as peak broadening, leaks, or loss of efficiency.<h4>The Early Days of HPLC Capillaries</h4>Anyone who has been working with HPLC for many years will probably remember the era of rigid 1/16" stainless steel tubing. Whenever PEEK tubing was unsuitable, stainless steel capillaries were the standard choice.Setting up a system back then was far from convenient. You first had to locate an appropriate length of tubing somewhere in a drawer full of capillaries. Usually, this meant finding a several-meter-long piece and cutting it yourself using a dedicated capillary cutter. After cutting, you needed to ensure the tubing end was smooth and undamaged, hoping that the previous person using the cutter had not accidentally crushed the inner diameter.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Standard_1_16_Capillary_and_needed_tools_and_the_drawer_we_all_know_pretty_well_943d7ed0b2.jpg" alt="KNAUER: Standard 1/16” Capillary and needed tools and the drawer we all know pretty well" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Standard_1_16_Capillary_and_needed_tools_and_the_drawer_we_all_know_pretty_well_943d7ed0b2.jpg 245w," sizes="100vw" width="1277" height="456">Once cut, the tubing often required additional preparation. You might need to deburr the capillary using a grater, shape it carefully with bending tools, and then finally install it using separate fittings and ferrules. And of course, everyone remembers the frustration of dropping a ferrule onto the laboratory floor, where it effectively disappeared forever.At the same time, laboratories often accumulated drawers filled with pre-assembled capillaries that had fixed fittings attached. Reusing these assemblies was risky. If the fitting geometry did not perfectly match the receiving port, users could unintentionally introduce dead volume into the system. The result was often degraded chromatographic performance, broad peaks, and frustrating troubleshooting sessions involving the entire laboratory team. In worse cases, excessive force could damage threads or receiving ports while attempting to stop leaks.During that period, assembling and connecting a complete HPLC system could easily consume half a day of work.<h4>The Improvement: Flexible and Pre-Cut Capillaries</h4>Later, laboratories gradually adopted pre-cut tubing and flexible capillaries, which represented a significant improvement over manually preparing stainless steel tubing.With these newer capillaries, users simply selected the correct length and installed the tubing directly. Flexible capillaries adapted naturally to the required shape, eliminating much of the manual bending and shaping work. Installation became faster, easier, and far less stressful.Still, these systems had limitations. Although the capillary itself improved, the fitting technology remained largely unchanged. Many of these capillaries were still effectively single-use components. If a connection was installed incorrectly because the tubing was not fully inserted into the receiving port before tightening, the result was often another unusable capillary destined for the growing “capillary drawer.” Sustainability was certainly not a strong point of these systems.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_A_typical_capillary_drawer_It_was_tricky_to_open_and_close_it_sometimes_530dd2087c.webp" alt="KNAUER: A typical capillary drawer. It was tricky to open and close it sometimes" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_A_typical_capillary_drawer_It_was_tricky_to_open_and_close_it_sometimes_530dd2087c.webp 208w," sizes="100vw" width="1024" height="768">Even with these advancements, system setup still required considerable time and effort.<h4>The Real Breakthrough: Modern Tool-Free Connection Systems</h4>A major breakthrough finally arrived when manufacturers introduced a completely redesigned fitting and capillary concept aimed specifically at simplifying UHPLC connections.These newer systems came with fittings pre-installed directly on the capillary. Unlike traditional ferrule-based systems, the fittings automatically adjusted to different port depths and sealed directly at the bottom of the port. This dramatically reduced the amount of torque required and eliminated the need for complicated installation techniques.Perhaps the biggest advantage was convenience. These capillaries could be connected and disconnected more than 100 times without tools, while remaining entirely finger-tight. And best of all, ferrules no longer disappeared onto the laboratory floor.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_The_little_things_that_do_the_click_3a0469d516.jpg" alt="KNAUER: The little things that do the “click”" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_The_little_things_that_do_the_click_3a0469d516.jpg 245w," sizes="100vw" width="1866" height="1022">The system I currently use includes an especially useful feature: a patented torque-limiting mechanism that prevents both under-tightening and over-tightening. Once the correct torque is reached, the fitting produces a tactile “click,” providing immediate feedback that the connection is secure and properly installed every time.<h4>From Half a Day to Fifteen Minutes</h4>Thanks to these modern connection systems and pre-configured startup kits, installing an HPLC or UHPLC system today is dramatically faster than it used to be.What once consumed hours of work can now be completed in less than 15 minutes. The improvements in usability, reproducibility, and sustainability over recent years have been remarkable.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Trouble_free_connection_systom_for_HPLC_and_UHPLC_e62b80154b.webp" alt="KNAUER: Trouble-free connection systom for HPLC and UHPLC" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Trouble_free_connection_systom_for_HPLC_and_UHPLC_e62b80154b.webp 221w," sizes="100vw" width="690" height="487"><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_Trouble_free_connection_systom_for_HPLC_and_UHPLC_2_71c24acdf5.webp" alt="KNAUER: Trouble-free connection systom for HPLC and UHPLC" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_Trouble_free_connection_systom_for_HPLC_and_UHPLC_2_71c24acdf5.webp 224w," sizes="100vw" width="1920" height="1339">And while today’s solutions already represent a huge leap forward, the evolution of HPLC connection technology is certainly not over yet. Looking ahead, it will be exciting to see how future innovations continue to simplify chromatography workflows even further.<blockquote>If you want to learn more about KNAUER´s capillaries, fittings and (U)HPLC system start-up kits and are interested in details and numbers, I highly recommend our brand <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/r75ev6rjivdbid246k60zrs3gjpvwdi5b8em_fad0f92fd9.pdf">new tech note "Benefits of KNAUER´s trouble-free analytical capillaries in terms of HPLC and UHPLC system performance</a>" and a visit to our <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/2402">products online</a>.</blockquote>

Let´s Connect a whole (U)HPLC system without tools and trouble

News from LabRulezLCMS Library - Week 21, 2026

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A unique company dedicated to serving science helping our customers drive productivity and accelerate results. Through our premier brands – Thermo Scientific, Applied Biosystems, Invitrogen, Fisher Scientific and Unity Lab Services.

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Based in Berlin, KNAUER is a medium-sized, owner-managed company that has been serving the sciences since 1962. We develop and manufacture scientific instruments of superior quality for liquid chromatography, including: Analytical HPLC/UHPLC, Preparative HPLC, Fast protein liquid chromatography (FPLC), Multi-column chromatography/Simulated moving bed (SMB) chromatography. High pressure dosing, in-line detection systems, and osmometry are additional business areas.

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KNAUER Eurospher II 100 C18P (U)HPLC Columns (USP L1)

KNAUER´s most unpolar C18P phase with excellent shape selectivity. It features polymeric, high efficiency endcapping of 99 % and 20 % carbon content, exceptional pH stability in the range of 1–12.

C18P columns with trifunctional modifications (USP L1): Hydrophobic and stearic interactionsKNAUER´s most unpolar C18P phase with excellent shape selectivity. It features polymeric, high efficiency endcapping of 99 % and 20 % carbon content, exceptional pH stability in the range of 1–12, and outstanding mechanical and chemical stability.<h5>Properties</h5><ul><li>Separation mechanism: Hydrophobic and stearic interaction</li><li>Ultra pure, spherical high performance HPLC phase based on silica gel</li><li>Unpolar, trifunctional C18 (Octadecyl) modification, high efficiency endcapping, with 20 % carbon content (~ 99 % endcapping)&nbsp;</li></ul><h5>Key features</h5><ul><li>KNAUER´s most unpolar C18 phase, polymeric, high efficiency endcapping of 99 % and 20 % carbon content, higher pH stability in the range of 1-12, outstanding mechanical and chemical stability, suited for analytical as well as semi preparative and preparative applications.</li></ul><h5>Recommended application areas</h5><ul><li>Alternative selectivity to standard C18; stationary phase in Eurospher II C18 family with the highest carbon load; fully endcapped; excellent shape selectivity and pH stability.</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_C18_P_f093126fa9.webp" alt="KNAUER: C18P" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_C18_P_f093126fa9.webp 245w," sizes="100vw" width="278" height="174"><blockquote>Tip: Not all C18 phases are comparable! When replacing an existing column, always have a close look at the characteristics like carbon load, pore size and specific surface area.</blockquote>Stationary phase Eurospher II 100 C18P is provided in these column types: analytical, semi-preparative and preparative.Eurospher II 100 C18P Analytical Columns are available with these parameters:<ul><li>Column length (mm): 50, 100, 150, 250</li><li>Column ID (mm): 2, 3, 4, 4.6</li><li>Particle size (µm): 2, 3, 5</li></ul>Eurospher II 100 C18P Semi-Preparative Standard Columns are available with these parameters:<ul><li>Column length (mm): 30, 50, 125, 150, 250</li><li>Column ID (mm): 8, 16</li><li>Particle size (µm): 5, 10</li></ul><h3>KNAUER (U)HPLC Columns</h3>Widest range of HPLC and UHPLC columns for all separation modes and applications.<ul><li>Safe Investment: Robust and reliable columns for attractive price&nbsp;</li><li>Separation Mode: Reversed Phase, HILIC, Normale</li><li>Simplicity: Easy purchase via our online store with fast delivery</li><li>Application: All-rounder for research or highly specialised for high-throughput analysis? We have the best column for your requirements.</li><li>Flexibility: Wide selection of column and particle sizes, for (U)HPLC applications up to 1000 bar</li></ul>KNAUER offers the perfect column for any small molecule separation. No matter which separation mode, molecule and the mobile phase, we offer a suitable (U)HPLC stationary phase. Choose from a wide range of modifications, particle sizes and column dimensions to suit your application. To elongate the lifetime of your column, you can choose to use a suitable precolumn.<figure class="image"><img style="aspect-ratio:787/674;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/KNAUER_KNAUER_U_HPLC_C8_and_C8_A_Columns_083c60f401.webp" alt="KNAUER: KNAUER (U)HPLC Columns" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_KNAUER_KNAUER_U_HPLC_C8_and_C8_A_Columns_083c60f401.webp 182w," sizes="100vw" width="787" height="674"></figure>

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News from LabRulezLCMS Library - Week 21, 2026

Let´s Connect a whole (U)HPLC system without tools and trouble

How to Cut HPLC Sample Preparation Time for Derivatizing Amino Acids