scholarly journals An HPLC tool for process monitoring: rare sugar D- psicose and D- fructose contents during the production through an enzymatic path

2020 ◽  
Vol 11 (1) ◽  
pp. 775-780 ◽  
Author(s):  
Sri Rama Krishna Surapureddi ◽  
Kunta Ravindhranath ◽  
Saritha Anthireddy
Keyword(s):  
Current Method ◽  
Raw Material ◽  
Reaction Monitoring ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Rare Sugar ◽  
Food Industries ◽  
On Line ◽  
Isocratic Hplc ◽  
Process Cost

D-Psicose/allulose, a rare sugar, is an essential raw material in the pharmaceutical and food industries. It is scantly found in nature and to meet its demand in industries, D-Psicose is generated enzymatically using D-fructose as a substrate.  In these conversations, it is important to monitor D-Psicose,  in order to control the process, impurities, optimize the reaction time and reduce the process cost The available analytical methods have their limitations in quantifying D-psicose and D-fructose mixtures. Hence there is a need for the development of a routine, sensitive, quick and precise analytical method for D-psicose production on-line monitoring of reaction mixer. In the present work, a simplified reverse phase HPLC technique is developed and validated for the quick reaction monitoring of D-psicose from D-fructose, during enzymatic conversation procedures. The analysis is conducted at different concentrations ranging from 0.05 % to 0.5 % of the standard solutions of the D-psicose and D-fructose, by using water and Acetonitrile (at a ratio of 20:80) as eluent with a flow rate of 1.0 mL/min on isocratic HPLC-RID system with an aminopropyl silane stationary phase [ZORBAX SIL 4.6 x 150 mm, 5 µm particle size column (USP-L8)]. The applicability of this method is illustrated in reaction monitoring, where D-fructose (substrate) is converted to D-psicose (product) in the presence of the enzyme:  D-Tagotose 3- epimerase. Separation of D-psicose and D-fructose is achieved within 8 minutes with a resolution ≥ 4 which is the key advantage for reaction monitoring and linearity is established with regression of ≥ 0.99.  Additionally, the current method uses a simple mobile phase, without any buffers. It can be used routinely for reaction monitoring.

scholarly journals Method Development for 4-Hydroxy-2-Methyl-N- (5-Methyl-2-Thiazolyl)-2H-1,2-Benzothiazine-3-Carboxamide-1,1-Dioxide by Reverse Phase HPLC: New Analytical Technique

2019 ◽  
pp. 1-10
Author(s):  
Husnul Maab ◽  
Hammad Yousaf ◽  
Syed Saeed ul Hassan ◽  
Umar Farooq
Keyword(s):  
Mobile Phase ◽  
Method Development ◽  
Raw Material ◽  
New Method ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Analytical Technique ◽  
Peak Areas ◽  
Standard Solution ◽  
Study Development

Purpose: In this study, development of a new analytical method for the evaluation of 4-Hydroxy-2-methyl-N-(5-methyl-2-thiazolyl)-2H-1,2-benzothiazine-3-carboxamide-1,1-dioxide (Meloxicam) by reverse phase HPLC was carried out. The basic aim of this research was to develop and validate a simple, precise, accurate and sensitive method for qualitative and quantitative analysis of Meloxicam in pharmaceutical raw material and its dosage forms. The existing reported method (BP) for the analysis of Meloxicam is potentiometric method which is an old, lengthy and tedious method. Methods: In the new method of reverse phase HPLC, C18 column was used while the mobile phase was acetonitrile and methanol (70:30). The flow rate of mobile phase was 0.6 ml/min and retention time was found to be 1.5min. Separately equal volume of standard solution and sample solutions in HPLC vials were injected in auto sampler compartment of HPLC in six replicates. Chromatogram and peak areas of Meloxicam in standard and sample solutions of different concentrations were recorded. Results: This method was later validated in different ways by which the calibration curve proved to be linear with linearity coefficient of 0.999 over the range of 100 to 600ppm. The precision was equivalent to 0.0003%. The LOD and LOQ were 0.0003ug/ml and 0.001ug/ml respectively. The system also showed accuracy over the range of 95 to 99%. Conclusion: Hence, this method proved to be an alternative to the existing reported method of potentiometric titration because the new method showed accuracy, reproducibility and sensitivity.

scholarly journals Determination of Montelukast Sodium in Raw Material and Solid Dosage Form Using Reverse Phase HPLC

2013 ◽  
Vol 25 (13) ◽  
pp. 7481-7484
Author(s):  
Syed Saeed-Ul-Hassan ◽  
Ahsan-Ul-Haq Ather ◽  
Muhammad Tayyab Ansari ◽  
Imran Tariq ◽  
Sabiha Karim
Keyword(s):  
Dosage Form ◽  
Raw Material ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Solid Dosage Form ◽  
Solid Dosage ◽  
Montelukast Sodium

A Simple Improved Method for the Measurement of Cyclosporin by Liquid-Liquid Extraction of Whole Blood and Isocratic HPLC

1988 ◽  
Vol 25 (5) ◽  
pp. 510-515 ◽  
Author(s):  
Stephen J Fletcher ◽  
Robby A Bacchus
Keyword(s):  
Whole Blood ◽  
Hplc Method ◽  
Improved Method ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Liquid Liquid Extraction ◽  
Routine Measurement ◽  
Ether Extraction ◽  
Acid Wash ◽  
Isocratic Hplc

The current HPLC methods of cyclosporin measurement have been reviewed and all aspects assessed. A simple isocratic C-18 reverse phase HPLC method with improved efficiency is described for the routine measurement of cyclosporin in whole blood. An alkaline ether extraction is followed by an acid wash, solvent evaporation and two hexane washes of the reconstituted extract. The turn-round time for a single sample is 1 h. Daily batches of up to 40 patient samples can be easily measured with this method. The results are compared with those from the Sandoz radioimmunoassay (RIA) method.

Measurement of elastin hydrolysis by reverse-phase HPLC with on-line post-colum derivatization

1989 ◽  
Vol 27 (5-6) ◽  
pp. 225-227 ◽  
Author(s):  
Th. A. Stein ◽  
J. R. Cohen ◽  
C. Mandell ◽  
L. Wise
Keyword(s):  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
On Line

Complete β-oxidation of valproate: cleavage of 3-oxovalproyl-CoA by a mitochondrial 3-oxoacyl-CoA thiolase

2002 ◽  
Vol 362 (3) ◽  
pp. 755-760 ◽  
Author(s):  
Margarida F. B. SILVA ◽  
Jos P. N. RUITER ◽  
Henk OVERMARS ◽  
Albert H. BOOTSMA ◽  
Albert H. van GENNIP ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Liver Mitochondria ◽  
Rat Liver Mitochondria ◽  
Tandem Ms ◽  
Carnitine Acetyltransferase ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
On Line ◽  
Hydrolysis Of

The β-oxidation of valproic acid (VPA; 2-n-propylpentanoic acid) was investigated in vitro in intact rat liver mitochondria incubated with 3H-labelled VPA. The metabolism of [4,5-3H2]VPA and [2-3H]VPA was studied by analysing the different acyl-CoA intermediates formed by reverse-phase HPLC with radiochemical detection. Valproyl-CoA, Δ2(E)-valproyl-CoA,3-hydroxyvalproyl-CoA and 3-oxovalproyl-CoA (labelled and non-labelled) were determined using continuous on-line radiochemical and UV detection. The formation of these intermediates was investigated using the two tritiated precursors in respiratory states 3 and 4. Valproyl-CoA was present at highest concentrations under both conditions. Two distinct labelled peaks were found, which were identified as 3H2O and [4,5-3H2]3-oxo-VPA. The formation of 3H2O strongly suggested that VPA underwent complete β-oxidation and that [4,5-3H2]3-oxo-VPA was formed by hydrolysis of the corresponding thioester. The hypothesis that 3-oxovalproyl-CoA undergoes thiolytic cleavage was investigated further. For this purpose a mito chondrial lysate was incubated with synthetic 3-oxovalproyl-CoA, carnitine and carnitine acetyltransferase for subsequent monitoring of the formation of propionylcarnitine and pentanoylcarnitine using electrospray ionization tandem MS. The detection of these compounds demonstrated unequivocally that the intermediate 3-oxovalproyl-CoA is a substrate of a mitochondrial thiolase, producing propionyl-CoA and pentanoyl-CoA, thus demonstrating the complete β-oxidation of VPA in the mitochondrion. Our data should lead to a re-evaluation of the generally accepted concept that the biotransformation of VPA by mitochondrial β-oxidation is incomplete.

COOH-terminally extended secretins are potent stimulants of pancreatic secretion

1999 ◽  
Vol 276 (4) ◽  
pp. G808-G816 ◽  
Author(s):  
Travis E. Solomon ◽  
John H. Walsh ◽  
Louis Bussjaeger ◽  
Yumei Zong ◽  
James W. Hamilton ◽  
...  
Keyword(s):  
Pancreatic Secretion ◽  
Cross Reactivity ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Mass Spectral ◽  
Tissue Extracts ◽  
Secretin Receptor ◽  
Ion Exchange Hplc ◽  
Isocratic Hplc ◽  
Stimulation Of

Posttranslational processing of preprosecretin generates several COOH-terminally extended forms of secretin and α-carboxyl amidated secretin. We used synthetic canine secretin analogs with COOH-terminal -amide, -Gly, or -Gly-Lys-Arg to examine the effects of COOH-terminal extensions of secretin on bioactivity and detection in RIA. Synthetic products were purified by reverse-phase and ion-exchange HPLC and characterized by reverse-phase isocratic HPLC and amino acid, sequence, and mass spectral analyses. Secretin and secretin-Gly were noted to coelute during reverse-phase HPLC. In RIA using eight different antisera raised against amidated secretin, COOH-terminally extended secretins had little or no cross-reactivity. Bioactivity was assessed by measuring pancreatic responses in anesthetized rats. Amidated canine and porcine secretins were equipotent. Secretin-Gly and secretin-Gly-Lys-Arg had potencies of 81 ± 9% ( P > 0.05) and 176 ± 13% ( P < 0.01), respectively, compared with amidated secretin, and the response to secretin-Gly-Lys-Arg lasted significantly longer. These data demonstrate that 1) amidated secretin and secretin-Gly are not separable under some chromatographic conditions, 2) current RIA may not detect bioactive COOH-terminally extended forms of secretin in tissue extracts or blood, and 3) the secretin receptor mediating stimulation of pancreatic secretion recognizes both amidated and COOH-terminally extended secretins.

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