SIELC’s Alltesta™ Analyzer is an affordable HPLC analytical system that provides both scientific and educational environments with a convenient, simple solution for measuring small and trace concentrations of various chemicals in products such as milk, baby formula, pet food, pills, household formulations, and more.No difficult sample preparation is necessary, and no prior experience in chromatography or analytical chemistry is required for the operator. Measurements typically take less than 5 minutes, and the Alltesta™ comes with a set of supplies sufficient for 200 analyses.<ul><li>Small foot-print, light-weight, can be used on a small table</li><li>No difficult sample preparation</li><li>Takes only a few minutes to start operating</li><li>No chemical experience is required for instrument user</li><li>User friendly, self explanatory cloud-based software</li><li>Produces easy-to-understand results within 5-10 minutes</li></ul><h4>Alltesta Gradient Automated HPLC Analyzer includes:</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1249">Alltesta™ Power Tower</a> (provides power and cloud connectivity)</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1244">Alltesta Mini Autosampler</a> (includes <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1245">Alltesta High-Pressure Switching Valve</a>)</li><li>Two <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1247">Alltesta™ Ultra High Pressure Mini Syringe Pumps</a> for gradient flow</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1248">Alltesta UV-Vis Detector</a></li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/SIELC_Alltesta_automaticky_HPLC_gradientovy_analyzator_2_6aec49522f.jpg" alt="Bioanalytic / SIELC: Alltesta Gradient Automated HPLC Analyzer" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_SIELC_Alltesta_automaticky_HPLC_gradientovy_analyzator_2_6aec49522f.jpg 107w," sizes="100vw" width="1969" height="2881"><h3>Application examples using Alltesta HPLC</h3><h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1242">Alltesta HPLC</a> Method for Berberine on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=243&amp;search=Primesep+B">Primesep B</a> by SIELC Technologies</h4>Berberine is a naturally occurring alkaloid found in several plants, including the roots, rhizomes, and stem bark of plants such as Berberis species (barberry), Coptis chinensis (goldthread), and Hydrastis canadensis (goldenseal).<ul><li>Medicinal Properties: Berberine has been used in traditional medicine for its various medicinal properties, including antimicrobial, anti-inflammatory, and antioxidant effects.</li><li>Blood Sugar Regulation: Some studies suggest that berberine may help regulate blood sugar levels and improve insulin sensitivity, making it of interest in diabetes research.</li><li>Cholesterol Management: It has been investigated for potential benefits in managing cholesterol levels.</li><li>Anticancer Properties: Berberine has also shown promise in preclinical studies for its potential anticancer properties.</li></ul>Berberine supplements are available and marketed for various health purposes. In traditional medicine, berberine-containing plants have been used for treating various conditions.<h5>HPLC Method</h5><ul><li>Column: Primesep B, 3.2 x 150 mm, 5 µm, 100 A</li><li>Mobile Phase: MeCN/H2O – 20/80%</li><li>Buffer: H3PO4 -0.1%</li><li>Flow Rate:&nbsp;0.5 ml/min</li><li>Detection:&nbsp;UV 275 nm</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_SIECL_Alltesta_HPLC_metoda_proanalyzu_Berberinu_na_kolone_Primesep_B_od_SIELC_Technologies_444d7c3d9c.jpg" alt="Bioanalytic / SIELC: Alltesta HPLC Method for Berberine on Primesep B by SIELC Technologies" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_SIECL_Alltesta_HPLC_metoda_proanalyzu_Berberinu_na_kolone_Primesep_B_od_SIELC_Technologies_444d7c3d9c.jpg 241w," sizes="100vw" width="775" height="500"><h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1242">Alltesta HPLC</a> Method for Prednisone, Methylprednisolone, Prednisolone on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=243&amp;search=Primesep+B">Primesep B</a> by SIELC Technologies</h4>All three compounds have potent anti-inflammatory effects, making them valuable in treating conditions where inflammation plays a role.Prednisone:<ul><li>Mechanism of Action: Prednisone is a synthetic corticosteroid that mimics the action of cortisol, a natural steroid produced by the adrenal glands. It primarily exerts its effects by binding to glucocorticoid receptors and regulating gene expression.</li><li>Medical Uses: It is commonly used to treat inflammatory conditions, autoimmune disorders, allergic reactions, and certain cancers.</li></ul>Prednisolone:<ul><li>Derived Form: Prednisolone is the active form of prednisone. When prednisone is ingested, the liver converts it into prednisolone.</li><li>Mechanism of Action: Similar to prednisone, it has anti-inflammatory and immunosuppressive properties.</li></ul>Methylprednisolone:<ul><li>Potency: Methylprednisolone is considered to be more potent than prednisone.</li><li>Medical Uses: It is used in various conditions, including severe allergic reactions, inflammatory conditions, and as a part of immunosuppressive regimens.</li></ul>HPLC Method<ul><li>Column:&nbsp;Primesep B, 3.2 x 150 mm, 5 µm, 100 A</li><li>Mobile Phase: MeCN/H2O – 35/35%</li><li>Buffer: H3PO4 -0.1%</li><li>Flow Rate:&nbsp;0.5 ml/min</li><li>Detection: UV 275 nm</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_SIELC_Alltesta_HPLC_metoda_pro_Prednison_Methylprednisolon_Prednisolon_na_kolone_Primesep_B_od_SIELC_Technologies_019e25f09c.jpg" alt="Bioanalytic / SIELC: Alltesta HPLC Method for Prednisone, Methylprednisolone, Prednisolone on Primesep B by SIELC Technologies" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_SIELC_Alltesta_HPLC_metoda_pro_Prednison_Methylprednisolon_Prednisolon_na_kolone_Primesep_B_od_SIELC_Technologies_019e25f09c.jpg 224w," sizes="100vw" width="775" height="540"><h4>More examples of Alltesta™-based HPLC methods are available in the LabRulez Library</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/HPLC_Method_for_Analysis_of_Aspirin_in_Alka_Seltzer_Extra_Strength_Effervescent_Aspirin_on_Primesep_100_Column_d33816ae83.pdf">HPLC Method for Analysis of Aspirin in Alka-Seltzer Extra Strength Effervescent Aspirin</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/HPLC_Method_for_Analysis_of_Phenylephrine_Hydrochloride_1_in_Equate_Nasal_Spray_on_Primesep_100_Column_02f2983c25.pdf">HPLC Method for Analysis of Phenylephrine Hydrochloride 1% in Equate – Nasal Spray</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/HPLC_Method_for_Analysis_of_Rockstar_Zero_Carbs_Energy_Drink_with_Caffeine_and_Taurine_on_Primesep_100_Column_c87a85a0b4.pdf">HPLC Method for Analysis of Rockstar Zero Carbs Energy Drink with Caffeine and Taurine</a></li></ul>See this cloud-based example of the 2-Naphthalenesulfonic acid, Allura red, Fast Green FCF, 1,5-Naphthalenedisulfonic acid, Tartrazine separation method <a target="_blank" rel="noopener noreferrer" href="https://chrom.app/share/#/samples/11972/injection/21307/?t=a992fefcdd3647b19f6bed81e182d3d8">here</a>.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/SIELC_SIELC_Alltesta_Gradient_Automated_Analyzer_example_of_the_2_Naphthalenesulfonic_acid_Allura_red_Fast_Green_FCF_1_5_Naphthalenedisulfonic_acid_Tartrazine_separation_b96e648d63.jpg" alt="Bioanalytic / SIELC: SIELC Alltesta™ Gradient Automated Analyzer - example of the 2-Naphthalenesulfonic acid, Allura red, Fast Green FCF, 1,5-Naphthalenedisulfonic acid, Tartrazine separation" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_SIELC_SIELC_Alltesta_Gradient_Automated_Analyzer_example_of_the_2_Naphthalenesulfonic_acid_Allura_red_Fast_Green_FCF_1_5_Naphthalenedisulfonic_acid_Tartrazine_separation_b96e648d63.jpg 245w," sizes="100vw" width="1914" height="1134">

Compact, cost-effective HPLC system - SIELC Alltesta

<h4 style="text-align:justify;">Background</h4>TDM of immunosuppressants is extremely important topic, especially when it comes to sample collection at home coupled with simple and fast sample preparation and analysis. The aims of the study covered check if optimization of ready-to-use kit for determination of Tacrolimus (TAC), Sirolimus (SIR), Everolimus (EVR) and Cyclosporin A (CSA) in venous blood can be successfully applied for VAMS-collected blood and preliminary venous blood to Mitra® (VB/M) correlation study.<blockquote>Check the link for pdf of the original poster in the <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/Poster_Immuno_3000px_RGB_352b008e9d.pdf">LabRulezLCMS library</a>!</blockquote><h4 style="text-align:justify;">Methods</h4>Sample preparation for venous blood was done according to the procedure provided with the kit (Chromsystems kit: 93000). Sample preparation for Mitra®-collected blood included additional sonication, vortexing and combining of internal standards with extraction buffer. Only reagents supplied by the manufacturer were used. All samples were analyzed on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=QTRAP+5500%2B">QTRAP 5500+</a> LC-MS/MS system (SCIEX). Data processing, quantitation and basic statistical analysis was done in SciexOS 3.3 software (SCIEX).<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Venous_Blood_Chromsystems_b496c1d87a.jpg" alt="Bioanalytic: Venous blood (Chromsystems procedure)" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Venous_Blood_Chromsystems_b496c1d87a.jpg 234w," sizes="100vw" width="1323" height="881"><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Mitra_optimized_procedure_e671781241.jpg" alt="Bioanalytic: Mitra - optimized procedure based on Chromsystems kit." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Mitra_optimized_procedure_e671781241.jpg 211w," sizes="100vw" width="1340" height="994"><figure class="image"><img style="aspect-ratio:2064/674;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Data_processing_quantitation_8529b80c50.jpg" alt="Bioanalytic: Data processing, quantitation and basic statistical analysis was done in SciexOS 3.3 software (SCIEX)." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Data_processing_quantitation_8529b80c50.jpg 245w," sizes="100vw" width="2064" height="674"></figure><h4 style="text-align:justify;">Results</h4>The optimized for 10µl and 20µl Mitra® samplers procedure met the validation criteria, including: reproducibility (%CV±15% and accuracy 85-115%), LLOQ (S/N≥10, calibrator 1), linearity (R≥0.995). LOD was not determined, however collected for 10 µl Mitra® sampler data shows that lower that 50 pg/ml concentration for each compound is achieved. The preliminary study of VB/M correlation, included 15 patients per compound where venous blood and 10 µl Mitra® samples were collected at the same time. Obtained VB/M factors were as follows: TAC=0.75, %CV=22.92%, SIR=0.77, %CV=36.10%, CSA=1.15, %CV=19.01%. No patients for EVR were tested during the study.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Tab_1_Validation_results_for_8c39ba1b30.jpg" alt="Bioanalytic: Tab. 1. Validation results for collected on 10 microL Mitra sampler lowest analytes concentrations provided with the kit (CAL 1, 4 replicates). LOD was not determined, however assitional analyses showed that lower that 50-100 pg/ml concentration for each compound can be achieved." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Tab_1_Validation_results_for_8c39ba1b30.jpg 245w," sizes="100vw" width="2679" height="925"><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Tab_Results_of_preliminary_1d5b77648e.jpg" alt="Bioanalytic: Tab. 2. Results of preliminary study for VB/M correlation included 15 paptients par compound where venous blood and 10 microL Mitra samples were collected at the same time. No patients for EVR were tested during this study." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_From_venous_blood_to_VAMS_adaptation_of_a_commercial_CE_IVD_assay_kit_for_the_determination_of_Tacrolimus_Sirolimus_Everolimus_and_Cyclosporin_A_Tab_Results_of_preliminary_1d5b77648e.jpg 245w," sizes="100vw" width="2168" height="1117"><h4 style="text-align:justify;">Conclusions</h4>Optimized for VAMS procedure allowed to achieve validation criteria with the use of kit reagents. Additional steps required for Mitra® sample preparation are simple, robust and automation friendly. The VB/M correlations revealed that the capillary blood concentration of TAC and SIR are lower than in venous blood contrary to concentration of CSA. This data may support the fact that TAC and SIR enters erythrocytes in about 90%, while CSA only in 50% and the capillary blood has higher amount of serum fraction. However, due to relatively small number of patients included in the study it needs further investigation on larger group of patients and deeper statistical analysis.<blockquote><h4 style="text-align:justify;">Authors</h4>Rafał Szewczyk1,4; Arkadiusz Kocur2,3, Agnieszka Czajkowska3; Karina Izdebska4; Maciej Stopa1,4. 1LabExperts Sp. z o.o., Gdańsk, Poland; 2Department of Drug Chemistry, Pharmaceutical and Biomedical Analysis, Faculty of Pharmacy, Medical University of Warsaw, Warsaw, Poland; 3Therapeutic Drug Monitoring, Clinical Pharmacokinetics and Toxicology Laboratory Unit, Department of Clinical Biochemistry, The Children’s Memorial Health Institute, Warsaw, Poland; 4Bioanalytic Sp. z o.o., Gdańsk, Poland</blockquote>

From venous blood to VAMS – adaptation of a commercial CE-IVD assay kit for the determination of Tacrolimus, Sirolimus, Everolimus and Cyclosporin A

<blockquote>Maciej Stopa 1,2, Julia Mironenka1, Rafał Szewczyk1,2, Anna Lenartowicz1, Adrian Soboń1,2, Katarzyna Krupczyńska-Stopa1,2, Andrzej Kwaśnica31. LabExperts sp. z o. o., Gdańsk, Poland; 2. <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/315">Bioanalytic</a> sp. z o. o., Gdańsk, Poland; 3. Lab4Tox sp. z o. o., Wrocław, Poland</blockquote><h4>Introduction</h4>The precision, reproducibility and trueness of the result in forensic analysis is crucial. The accuracy of sample preparation during the psychoactive compounds quantitation is one of the key aspects. Automatic sample preparation minimize mistakes caused by poor performance of the analysis, discrepancies between analysts habits or different days of analysis. Accurate analytes detection relies on efficient extraction, low detection limits and maintaining the efficiency and cleanliness of LC-MS/MS system. Liquid-liquid extraction, by default, are less amenable for automation, and may be challenging because of large amount of sample preparation steps. This work shows the effective translation from manual method to semi-automated procedure of determination of 34 psychoactive compounds from human plasma using LC-MS/MS method.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_1_Semi_automated_sample_07f1c50562.jpg" alt="Bioanalytic: Figure 1 - Semi-automated sample preparation protocol using Hudson SOLO automatic pipette station with Hudson SOLO Soft and SoftLink V protocol Editor." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_1_Semi_automated_sample_07f1c50562.jpg 123w," sizes="100vw" width="805" height="1021"><h4>Methods</h4>Liquid-liquid extraction is sufficient method to determine the amount of psychoactive compounds in human plasma. &nbsp;Sample preparation procedure developed using Hudson SOLO automatic pipette station with Hudson SOLO Soft and SoftLink V Protocol Editor include deproteinization and extraction, dissolution and transfer to a plate compatible with an LC autosampler in 100 minutes. Chromatography run was performed using ExionAC equipped with Fortis H2O C18 column. The method was run in reverse phase mode, using as mobile phases acidified with formic acid both, water and acetonitrile. Mass spectrometry was performed using &nbsp;ZenoTOF 7600 (SCIEX), with following conditions: positive electrospray scheduled MRMhr mode acquiring full MS/MS spectra. The data analysis including compound identification and quantification was performed with SciexOS 3.3 software.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Table_1_Part_A_The_comparison_136f3d2667.jpg" alt="Bioanalytic: Table 1. Part A - The comparison of manual vs-semi automated sample preparation precision; Part B: The results of method of semi-automated sample preparation." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Table_1_Part_A_The_comparison_136f3d2667.jpg 129w," sizes="100vw" width="1008" height="1217"><h4>Preliminary data</h4>Blood is&nbsp; easy to obtain information-rich matrix. It have only one disadvantage – it cannot be easily analyzed with LC-MS/MS. Preparation of a multimethod for psychoactive compounds determination which &nbsp;characterized by a different chemical properties requires complex and strict sample preparation to achieve accurate detection and compound presence confirmation. Unfortunately, in complex extraction steps are difficult to postpone due to various activities that has to be undertaken including – mixing, centrifugation, sample aspirations and evaporation. In this work, we show the possibility to transfer steps of aspiration, dilution, heating and mixing during the sample preparation on an automatic high precision pipette station. Other steps including centrifugation and evaporation were done manually. Beside the shortening of sample preparation time, which allows to perform the analysis of 96 samples in less than 2 hours, the results are subjected to less preparation error. Such configurations allow a substantial increase of sample throughput and convenience when compared to standard protocols. As a result of this translation, better repeatability (%CV ≤ 15%), maintaining a wide range of linearity (LLOQ-50 ng/ml) which met the validation criteria (R ≥ 0.995) of the method was achieved. An additional advantages of using automation – the analyst’s exposure to blood obtained from contractors is reduced. Also, automated protocol minimize the risk of losing valuable samples avoiding cases of human error.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_2_Chromatographic_separation_1f0e513084.jpg" alt="Bioanalytic: Figure 2: Chromatographic separation of analytes included in multi-method of psychoactive compounds quantitative determinantion [c=25 ng/ml]." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_2_Chromatographic_separation_1f0e513084.jpg 245w," sizes="100vw" width="2841" height="923"><h4>Novel aspect</h4>Automated method offers excellent repeatability and high accuracy of psychoactive compounds determination in human blood.

How to improve the precision of analysis – semi-automated sample preparation for psychoactive compounds quantification in human plasma

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How to improve the precision of analysis – semi-automated sample preparation for psychoactive compounds quantification in human plasma

From venous blood to VAMS – adaptation of a commercial CE-IVD assay kit for the determination of Tacrolimus, Sirolimus, Everolimus and Cyclosporin A

Compact, cost-effective HPLC system - SIELC Alltesta

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