scholarly journals PAT for Continuous Chromatography Integrated into Continuous Manufacturing of Biologics towards Autonomous Operation

Processes ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 472
Author(s):  
Florian Lukas Vetter ◽  
Steffen Zobel-Roos ◽  
Jochen Strube
Keyword(s):  
Molecular Weight ◽  
Zero Point ◽  
Test System ◽  
Target Component ◽  
Array Detector ◽  
Continuous Manufacturing ◽  
Fluorescence Detector ◽  
Autonomous Operation ◽  
Side Component ◽  
Process Disturbances

This study proposes a reliable inline PAT concept for the simultaneous monitoring of different product components after chromatography. The feed for purification consisted of four main components, IgG monomer, dimer, and two lower molecular weight components of 4.4 kDa and 1 kDa molecular weight. The proposed measurement setup consists of a UV–VIS diode-array detector and a fluorescence detector. Applying this system, a R2 of 0.93 for the target component, a R2 of 0.67 for the dimer, a R2 of 0.91 for the first side component and a R2 of 0.93 for the second side component is achieved. Root mean square error for IgG monomer was 0.027 g/L, for dimer 0.0047 g/L, for side component 1 0.016 g/L and for the side component 2 0.014 g/L. The proposed measurement concept tracked component concentration reliably down to 0.05 g/L. Zero-point fluctuations were kept within a standard deviation of 0.018 g/L for samples with no IgG concentration but with side components present, allowing a reliable detection of the target component. The main reason inline concentration measurements have not been established yet, is the false-positive measurement of target components when side components are present. This problem was eliminated using the combination of fluorescence and UV–VIS data for the test system. The use of this measurement system is simulated for the test system, allowing an automatic fraction cut at 0.05 g/L. In this simulation a consistent yield of >99% was achieved. Process disturbances for processed feed volume, feed purity and feed IgG concentration can be compensated with this setup. Compared to a timed process control, yield can be increased by up to 12.5%, if unexpected process disturbances occur.

scholarly journals Single-Laboratory Validation of a High-Performance Liquid Chromatographic-Diode Array Detector-Fluorescence Detector/Mass Spectrometric Method for Simultaneous Determination of Water-Soluble Vitamins in Multivitamin Dietary Tablets

2009 ◽  
Vol 92 (2) ◽  
pp. 680-688 ◽  
Author(s):  
Pei Chen ◽  
Renata Atkinson ◽  
Wayne R Wolf
Keyword(s):  
Formic Acid ◽  
Dietary Supplements ◽  
High Performance ◽  
Array Detector ◽  
Diode Array ◽  
Diode Array Detector ◽  
Fluorescence Detector ◽  
Liquid Chromatographic ◽  
Single Laboratory

Abstract The purpose of this study was to develop a single-laboratory validated (SLV) method using high-performance liquid chromatography with different detectors diode array detector (DAD); fluorescence detector (FLD); and mass spectrometry (MS) for determination of 7 B-complex vitamins (B1-thiamin, B2-riboflavin, B3-nicotinamide, B6-pyridoxine, B9-folic acid, pantothenic acid, and biotin) and vitamin C in multivitamin/multimineral dietary supplements. The method involves the use of a reversed-phase octadecylsilyl column (4 m, 250 2.0 mm id) and a gradient mobile phase profile. Gradient elution was performed at a flow rate of 0.25 mL/min. After a 5 min isocratic elution at 100 A (0.1 formic acid in water), a linear gradient to 50 A and 50 B (0.1 formic acid in acetonitrile) at 15 min was employed. Detection was performed with a DAD as well as either an FLD or a triple-quadrupole MS detector in the multiple reaction monitoring mode. SLV was performed using Standard Reference Material (SRM) 3280 Multivitamin/Multimineral Tablets, being developed by the National Institute of Standards and Technology, with support by the Office of Dietary Supplements of the National Institutes of Health. Phosphate buffer (10 mM, pH 2.0) extracts of the NIST SRM 3280 were analyzed by the liquid chromatographic (LC)-DAD-FLD/MS method. Following extraction, the method does not require any sample cleanup/preconcentration steps except centrifugation and filtration.

scholarly journals Depth of maximum of air-shower profiles: testing the compatibility of measurements performed at the Pierre Auger Observatory and the Telescope Array experiment

2019 ◽  
Vol 210 ◽  
pp. 01009
Author(s):  
Alexey Yushkov ◽  
Jose Bellido ◽  
John Belz ◽  
Vitor de Souza ◽  
William Hanlon ◽  
...  
Keyword(s):  
Quantitative Comparison ◽  
Pierre Auger Observatory ◽  
Array Detector ◽  
Air Showers ◽  
Fluorescence Detector ◽  
Telescope Array ◽  
Longitudinal Development ◽  
Air Shower ◽  
Direct Observations ◽  
Systematic Uncertainties

At the Pierre Auger Observatory and the Telescope Array, the measurements of depths of maximum of airshower profiles, Xmax, are performed using direct observations of the longitudinal development of showers with the help of the fluorescence telescopes. Though the same detection technique is used at both installations, the straightforward comparison of the characteristics of the measured Xmax distributions is not possible due to the different approaches to the analysis of the recorded events. In this work, the Auger – Telescope Array composition working group presents a technique to compare the Xmax measurements from the Auger Observatory and the Telescope Array. Applying this technique the compatibility of the first two moments of the measured Xmax distributions is qualitatively tested for energies 1018.2 eV < E < 1019.0 eV using the recently published Telescope Array data from the Black Rock Mesa and Long Ridge fluorescence detector stations. For a quantitative comparison, simulations of air showers with EPOS-LHC, folded with effects of the Telescope Array detector, are required along with the inclusion in the analysis of the systematic uncertainties in the measurements of Xmax and the energies of the events.

scholarly journals Synchronous High-Performance Liquid Chromatography with a Photodiode Array Detector and Mass Spectrometry for the Determination of Citrinin, Monascin, Ankaflavin, and the Lactone and Acid Forms of Monacolin K in Red Mold Rice

2011 ◽  
Vol 94 (1) ◽  
pp. 179-190 ◽  
Author(s):  
Cheng-Lun Wu ◽  
Yao-Haur Kuo ◽  
Chun-Lin Lee ◽  
Ya-Wen Hsu ◽  
Tzu-Ming Pan
Keyword(s):  
High Performance ◽  
Array Detector ◽  
Monacolin K ◽  
Fluorescence Detector ◽  
Photodiode Array Detector ◽  
Photodiode Array ◽  
Red Mold Rice ◽  
Product Red ◽  
Yellow Pigments

Abstract The Monascus fermentation product red mold rice (RMR) has been found to contain the cholesterol-lowering agent monacolin K (MK) in both its lactone (MKL) and acid (MKA) forms and the mycotoxin citrinin (CT). The yellow pigments in RMR, namely, monascin (MS) and ankaflavin (AK), have been reported to exhibit antimetastatic and antiangiogenic activities. Currently, MK and these yellow pigments are usually detected in RMR by different analytical methods that are inconvenient, expensive, and time-consuming. The goal of this study was to establish a rapid, synchronous analytical method for determination of the MKA, MKL, MS, AK, and CT levels in RMR. MKA, MKL, MS, AK, and CT were extracted by the same extraction method, then separated by RP-HPLC with a C18 column. The effluent from the column was passed through a photodiode array detector and then introduced directly into a fluorescence detector. The results showed that high recovery rates of MKA, MKL, MS, AK, and CT are possible if RMR powder is extracted with 75% ethanol (10 mL) at 80°C for 30 min. With regard to the optimal conditions of the HPLC, the peaks of MKA, MKL, MS, AK, and CT can be clearly separated from any noise peaks by isocratic elution with a mobile phase comprising 0.05% trifluoroacetic acid in acetonitrile–water (62.5 + 37.5, v/v).

scholarly journals Analytical procedures for the determination of estrogen compounds in a surface water reservoir in Southeast Brazil

2021 ◽  
Vol 46 (1) ◽  
pp. 41-51
Author(s):  
Thayná Aparecida Cais ◽  
Alley Michael da Silva Procópio ◽  
Márcia Matiko Kondo ◽  
Flávio Soares Silva ◽  
Sandro José De Andrade
Keyword(s):  
Surface Water ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Sewage Treatment Plant ◽  
Treatment Plant ◽  
Water Reservoir ◽  
Array Detector ◽  
Sample Point ◽  
Fluorescence Detector ◽  
Analytical Methodology

In this study, an analytical methodology was validated to determine and quantify four estrogen hormones using high-performance liquid chromatography (HPLC) with detections by diode array detector (DAD) and by fluorescence detector (FLD). For validation of the method, the following parameters were evaluated: linearity, selectivity, precision, accuracy, limit of detection (LOD), limit of quantification (LOQ) and robustness. Environmental samples were preconcentrated using solid phase extractions and for that, an experimental design was planned to determine the best recovery conditions by varying cartridge types, flow of eluent, pH of the samples, and eluting solvent. Five surface water sampling campaigns were carried out in five different sites of Furnas Reservoir over the months of December 2015 and May 2016. Sample point 1 was located near the sewage treatment plant of the city of Alfenas - MG, while sample point 5 was the most distant from this location. All estrogens, except for E1, were found in all water samples of at least one of the sampling sites. The concentrations of E3, E2 and EE2 ranged from 11-366, 63-422 and 75-9998 ng L-1, respectively. These results are consistent with several studies published in the scientific literature.

scholarly journals Fast and Flexible mRNA Vaccine Manufacturing as a Solution to Pandemic Situations by Adopting Chemical Engineering Good Practice—Continuous Autonomous Operation in Stainless Steel Equipment Concepts

Processes ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1874
Author(s):  
Axel Schmidt ◽  
Heribert Helgers ◽  
Florian Lukas Vetter ◽  
Alex Juckers ◽  
Jochen Strube
Keyword(s):  
Stainless Steel ◽  
Chemical Engineering ◽  
Process Analytical Technology ◽  
Good Practice ◽  
Operation Mode ◽  
Continuous Manufacturing ◽  
Efficient Operation ◽  
Messenger Ribonucleic Acid ◽  
Autonomous Operation ◽  
Analytical Technology

SARS-COVID-19 vaccine supply for the total worldwide population has a bottleneck in manufacturing capacity. Assessment of existing messenger ribonucleic acid (mRNA) vaccine processing shows a need for digital twins enabled by process analytical technology approaches in order to improve process transfer for manufacturing capacity multiplication, a reduction in out-of-specification batch failures, qualified personal training for faster validation and efficient operation, optimal utilization of scarce buffers and chemicals and speed-up of product release by continuous manufacturing. In this work, three manufacturing concepts for mRNA-based vaccines are evaluated: Batch, full-continuous and semi-continuous. Technical transfer from batch single-use to semi-continuous stainless-steel, i.e., plasmid deoxyribonucleic acid (pDNA) in batch and mRNA in continuous operation mode, is recommended, in order to gain: faster plant commissioning and start-up times of about 8–12 months and a rise in dose number by a factor of about 30 per year, with almost identical efforts in capital expenditures (CAPEX) and personnel resources, which are the dominant bottlenecks at the moment, at about 25% lower operating expenses (OPEX). Consumables are also reduceable by a factor of 6 as outcome of this study. Further optimization potential is seen at consequent digital twin and PAT (Process Analytical Technology) concept integration as key-enabling technologies towards autonomous operation including real-time release-testing.

scholarly journals Comparison of DAD and FLD Detection for Identification of Selected Bisphenols in Human Breast Milk Samples and Their Quantitative Analysis by LC-MS/MS

2020 ◽  
Vol 103 (4) ◽  
pp. 1029-1042
Author(s):  
Tomasz Tuzimski ◽  
Szymon Szubartowski ◽  
Renata Gadzała-Kopciuch ◽  
Andrzej Miturski ◽  
Monika Wójtowicz-Marzec ◽  
...  
Keyword(s):  
Breast Milk ◽  
Solid Phase ◽  
Array Detector ◽  
Human Breast Milk ◽  
Human Breast ◽  
Bisphenol S ◽  
Fluorescence Detector ◽  
Detection Techniques ◽  
Milk Samples

Abstract Background Determination of bisphenols released from packaging material is undoubtedly a difficult and tricky task, requiring the chemical analyst to develop an individual approach to obtain reliable analytical information. Objective QuECHERS (Quick, Easy, Cheap, Effective, Rugged, and Safe)/dispersive solid-phase extraction (d-SPE) technique and high performance liquid chromatography (HPLC) coupled with modern detection techniques such as diode-array detector (DAD), fluorescence detector (FLD) or tandem mass spectrometry (MS/MS) for the determination of bisphenols such as bisphenol A (BPA), bisphenol S (BPS), bisphenol F (BPF), bisphenol B (BPB), 2-[[4-[2-[4-(Oxiran-2-ylmethoxy)phenyl]propan-2yl]phenoxy] methyl]oxirane (BADGE), 3-[4-[2-[4-(Oxiran-2-ylmethoxy)phenyl]propan-2-yl]phenoxy]propane-1,2-diol (BADGE*H2O), 3-[4-[2-[4-(2,3-Dihydroxypropoxy)phenyl]propan-2-yl]phenoxy]propane-1,2-diol (BADGE*2H2O), 1-Chloro-3-[4-[2-[4-(3-chloro-2-hydroxypropoxy)phenyl] propan-2-yl]phenoxy]propan-2-ol (BADGE*2HCl) in human breast milk samples have been performed. Methods For the analysis of total analytes, prior to the extraction with acetonitrile, a deconjugation step was implemented in a tube by adding 1 mL of the enzymatic solution with the β-Glucuronidase to 5 mL of sample. The mix was homogenized and incubated for 17 h at 37°C. Ten milliliters of acetonitrile, and a QuEChERS salt packet with 4 g anhydrous MgSO4 and 1 g NaCl were added. During the d-SPE step the extract was transferred into tube with 30 mg Z-Sep and 50 mg PSA (and also 150 mg MgSO4 for LC-MS/MS analysis). MeOH–water (20:80, v/v) were added to the dry residue and the extract was reconstituted in 150 µL (25-fold analytes pre-concentration is achieved). Next bisphenols were identified by HPLC-DAD-FLD and quantified by LC-MS/MS equipment. Conclusions During the bisphenols HPLC-DAD-FLD analysis, from 6 min a reinforcement of 15 was used, which allowed analytes to be identified at 750 pg/mL. Application of LC-MS/MS allowed quantification of bisphenols in the range from 2.12 to 116.22 ng/mL in a total 27 human breast milk samples. Highlights First QuEChERS/d-SPE coupled with HPLC-DAD-FLD or LC-MS/MS method for the quantification of bisphenols and its analogues in breast milk Faster and cheaper alternative to traditional extraction methods The method was applied for the first biomonitoring of bisphenols and its analogues in breast milk.

scholarly journals Reaction of plasmic degradation products of fibrinogen in the radioimmunoassay of human fibrinopeptide A

Blood ◽  
1975 ◽  
Vol 45 (6) ◽  
pp. 757-768 ◽  
Author(s):  
AZ Budzynski ◽  
VJ Marder ◽  
S Sherry
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Venous Thrombosis ◽  
Degradation Products ◽  
Rabbit Antiserum ◽  
Amino Acid Position ◽  
Test System ◽  
Fibrinopeptide A ◽  
Terminal Portion ◽  
Large Molecular Weight

Abstract A radioimmunoassay (RIA) technique has been devised for the measurement of human fibrinopeptide A (FPA). The system utilizes rabbit antiserum to native human FPA and a synthetic fibrinopeptide, with tyrosine substituted for phenylanine in amino acid position 8. The test detects native human FPA at a concentration of 0.1 ng/ml, but does not cross react with human fibrinopeptide B or with fibrinopeptides A from canine, porcine, or bovine fibrinogen. Fibrinogen and chemical or plasmic degradation products with 2 moled of FPA per mole react fully in this test system. This includes the large-molecular-weight intermediate fragments X and Y and the NH2-terminal disulfide knot, and indicates that this antibody recognizes and reacts with FPA in the presence of the contiguous peptide structures present in fibrinogen. Fragment E, which is derived from the NH-2-terminal portion of fibrinogen, loses most of its FPA content after its liberation from its precursor derivative and reacts to a lesser extent in the RIA than do fragments X and Y. This correlated with the recovery of FPA-positive material from ultrafilitrates of extensive but not partial plasmic digests of fibrinogen. Although FPA immunoreactivity liberated from fibrinogen does not necessarily reflect thrombin activity and/or fibrin formation, only extensive plasmic degradation yields peptide material which reacts in this RIA system. This should not be a serious limitation to the application of the RIA in the detection of venous thrombosis.

scholarly journals Determination of Zearalenone Content in Cereals and Feedstuffs by Immunoaffinity Column Coupled with Liquid Chromatography

2001 ◽  
Vol 84 (5) ◽  
pp. 1453-1459 ◽  
Author(s):  
Bela Fazekas ◽  
Andrea Tar
Keyword(s):  
Liquid Chromatography ◽  
Recovery Rate ◽  
Limit Of Detection ◽  
Poultry Feed ◽  
Array Detector ◽  
Immunoaffinity Column ◽  
Uv Spectrum ◽  
Fluorescence Detector ◽  
Immunoaffinity Columns

Abstract The zearalenone content of maize, wheat, barley, swine feed, and poultry feed samples was determined by immunoaffinity column cleanup followed by liquid chromatography (IAC–LC). Samples were extracted in methanol–water (8 + 2, v/v) solution. The filtered extract was diluted with distilled water and applied to immunoaffinity columns. Zearalenone was eluted with methanol, dried by evaporation, and dissolved in acetonitrile–water (3 + 7, v/v). Zearalenone was separated by isocratic elution of acetonitrile–water (50 + 50, v/v) on reversed-phase C18 column. The quantitative analysis was performed by fluorescence detector and confirmation was based on the UV spectrum obtained by a diode array detector. The mean recovery rate of zearalenone was 82–97% (RSD, 1.4–4.1%) on the original (single-use) immunoaffinity columns. The limit of detection of zearalenone by fluorescence was 10 ng/g at a signal-to-noise ratio of 10:1 and 30 ng/g by spectral confirmation in UV. A good correlation was found (R2 = 0.89) between the results obtained by IAC–LC and by the official AOAC–LC method. The specificity of the method was increased by using fluorescence detection in parallel with UV detection. This method was applicable to the determination of zearalenone content in cereals and other kinds of feedstuffs. Reusability of immunoaffinity columns was examined by washing with water after sample elution and allowing columns to stand for 24 h at room temperature. The zearalenone recovery rate of the regenerated columns varied between 79 and 95% (RSD, 3.2–6.3%). Columns can be regenerated at least 3 times without altering their performance and without affecting the results of repeated determinations.

scholarly journals Process Analytical Technology for Precipitation Process Integration into Biologics Manufacturing towards Autonomous Operation—mAb Case Study

Processes ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 488
Author(s):  
Lara Julia Lohmann ◽  
Jochen Strube
Keyword(s):  
Process Control ◽  
Real Time ◽  
Process Analytical Technology ◽  
Measurement Techniques ◽  
Array Detector ◽  
Precipitation Process ◽  
Suitable Model ◽  
Process Data ◽  
Autonomous Operation ◽  
Analytical Technology

The integration of real time release testing into an advanced process control (APC) concept in combination with digital twins accelerates the process towards autonomous operation. In order to implement this, on the one hand, measurement technology is required that is capable of measuring relevant process data online, and on the other hand, a suitable model must be available to calculate new process parameters from this data, which are then used for process control. Therefore, the feasibility of online measurement techniques including Raman-spectroscopy, attenuated total reflection Fourier transformed infrared spectroscopy (ATR-FTIR), diode array detector (DAD) and fluorescence is demonstrated within the framework of the process analytical technology (PAT) initiative. The best result is achieved by Raman, which reliably detected mAb concentration (R2 of 0.93) and purity (R2 of 0.85) in real time, followed by DAD. Furthermore, the combination of DAD and Raman has been investigated, which provides a promising extension due to the orthogonal measurement methods and higher process robustness. The combination led to a prediction for concentration with a R2 of 0.90 ± 3.9% and for purity of 0.72 ± 4.9%. These data are used to run simulation studies to show the feasibility of process control with a suitable digital twin within the APC concept.

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