Sensitive UHPLC-MS/MS technique for monitoring Levothyroxine (T4) in human serum against endogenous thyroxin level

Background: Levothyroxine is a synthetic thyroid hormone that is chemically identical to Thyroxine (T4), which is secreted by the follicular cells of the thyroid gland. Levothyroxine is used to treat deficiency of thyroid hormone and to prevent the recurrence of thyroid cancer. Levothyroxine is present endogenously in human body. Method: It requires treated matrix for the preparation of calibration curve standard and quality control samples. The method was developed using LC-MS/MS and validated in human charcoal stripped serum. Charcoal stripped matrix was used for the preparation of Calibration curve standards and Quality control samples. Method involves Solid-Phase Extraction technique. Levothyroxine D3 used as an internal standard (ISTD). Result: Chromatographic separation was achieved using reversed phase analytical column Gemini NX-C18 110Å, 3µm (50x3.6) mm. Mobile phase consisted of acetonitrile and water in a ratio of 70:30 with 150µL of formic acid in 1000 mL of mobile phase. Mobile phase achieved shorter run-time of 0.9 minute due to use of Ultra-high performance liquid chromatography (UHPLC). Positive electro-spray ionization technique detected MRM ion pair transitions 777.60→731.65 for Levothyroxine and 780.70→734.6 for Levothyroxine- D3 (ISTD) were used. AB SCIEX Triple Quad™ API-4000 LC-MS/MS system and the bioanalytical method with 10ng/mL as limit of quantification has been applied successfully to pharmacokinetics studies. Conclusion: Chromatographic separation was achieved using reversed phase analytical column Gemini NX-C18 110Å, 3µm (50x3.6) mm. Mobile phase consisted of acetonitrile and water in a ratio of 70:30 with 150µL of formic acid in 1000 mL of mobile phase. Mobile phase achieved shorter run-time of 0.9 minute due to use of Ultra-high performance liquid chromatography (UHPLC). Positive electro-spray ionization technique detected MRM ion pair transitions 777.60→731.65 for Levothyroxine and 780.70→734.6 for Levothyroxine- D3 (ISTD) were used. AB SCIEX Triple Quad™ API-4000 LC-MS/MS system and the bioanalytical method with 10ng/mL as limit of quantification has been applied successfully to pharmacokinetics studies.

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Quantification of Histidine-Containing Dipeptides in Dolphin Serum Using a Reversed-Phase Ion-Pair High-Performance Liquid Chromatography Method

Separations ◽

10.3390/separations8080128 ◽

2021 ◽

Vol 8 (8) ◽

pp. 128

Author(s):

Momochika Kumagai ◽

Sanae Kato ◽

Nanami Arakawa ◽

Mika Otsuka ◽

Takahisa Hamano ◽

...

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Potassium Dihydrogen Phosphate ◽

Reversed Phase ◽

Ion Pair ◽

Dihydrogen Phosphate ◽

Limit Of Quantification ◽

Chromatography Method ◽

Relative Standard

The quantification of histidine-containing dipeptides (anserine, carnosine, and balenine) in serum might be a diagnostic tool to assess the health condition of animals. In this study, an existing reversed-phase ion-pair high-performance liquid chromatography (HPLC)–ultraviolet detection method was improved and validated to quantify serum anserine, carnosine, and balenine levels in the dolphin. The serum was deproteinized with trichloroacetic acid and directly injected into the HPLC system. Chromatographic separation of the three histidine-containing dipeptides was achieved on a TSK–gel ODS-80Ts (4.6 mm × 150 mm, 5 µm) analytical column using a mobile phase of 50 mmol/L potassium dihydrogen phosphate (pH 3.4) containing 6 mmol/L 1-heptanesulfonic acid and acetonitrile (96:4). The standard curve ranged from 0.1 µmol/L to 250 µmol/L. The average accuracy of the intra- and inter-analysis of anserine, carnosine, and balenine was 97–106%. The relative standard deviations of total precision (RSDr) of anserine, carnosine, and balenine in dolphin serum were 5.9%, 4.1%, and 2.6%, respectively. The lower limit of quantification of these compounds was 0.11–0.21 µmol/L. These results indicate that the improved method is reliable and concise for the simultaneous determination of anserine, carnosine, and balenine in dolphin serum, and may be useful for evaluation of health conditions in dolphins. Furthermore, this method can also be applied to other biological samples.

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Matrine Determination in Compound Sophora Alopecuroides Suppositories by High-Performance Liquid Chromatography

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.634-638.1001 ◽

2013 ◽

Vol 634-638 ◽

pp. 1001-1004

Author(s):

Hui Xiang Hu ◽

Xuan Zhang ◽

Zhimin Zhang

Keyword(s):

Quality Control ◽

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Mobile Phase ◽

High Performance ◽

Reversed Phase ◽

Average Recovery ◽

Sophora Alopecuroides ◽

Rp Hplc

The determination of matrine in compound sophora alopecuroides suppositories was carried out by reversed-phase high-performance liquid chromatography(RP-HPLC). Elite ODS C18(250mm×4.5mm, 5µm) column was used and acetonitrile -anhydrous ethanol-water-triethy lamine (15:22.5:62.5:0.01) was selected as the mobile phase. Meanwhile the detection wavelength 215nm was used. The linearity of matrine was in the range of 0.74-5.18µg，r=0.9991，and the average recovery was 98.52%，RSD was 1.54%. The method was proved to be simple, accurate, sensitive and reliable. It can be used for the quality control of compound sophora alopecuroides suppositories.

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Separation and Determination of Some Aromatic Sulfones by Reversed-Phase High-Performance Liquid Chromatography

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19940569 ◽

1994 ◽

Vol 59 (3) ◽

pp. 569-574 ◽

Cited By ~ 1

Author(s):

Josef Královský ◽

Marta Kalhousová ◽

Petr Šlosar

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Mobile Phase ◽

High Performance ◽

Reversed Phase ◽

Uv Spectra ◽

Optimum Conditions ◽

Linear Calibration ◽

Technical Grade

The reversed-phase high-performance liquid chromatography of some selected, industrially important aromatic sulfones has been investigated. The chromatographic behaviour of three groups of aromatic sulfones has been studied. The optimum conditions of separation and UV spectra of the sulfones and some of their hydroxy and benzyloxy derivatives are presented. The dependences of capacity factors vs methanol content in mobile phase are mentioned. The results obtained have been applied to the quantitative analysis of different technical-grade samples and isomer mixtures. For all the separation methods mentioned the concentration ranges of linear calibration curves have been determined.

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4-Nitrophenol in 4-nitrophenyl phosphate, a substrate for alkaline phosphatase, as measured by paired-ion high-performance liquid chromatography.

Clinical Chemistry ◽

10.1093/clinchem/23.12.2288 ◽

1977 ◽

Vol 23 (12) ◽

pp. 2288-2291 ◽

Cited By ~ 4

Author(s):

P H Culbreth ◽

I W Duncan ◽

C A Burtis

Keyword(s):

Alkaline Phosphatase ◽

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Flow Rate ◽

Mobile Phase ◽

High Performance ◽

Reversed Phase ◽

Nitrophenyl Phosphate ◽

Phase Column ◽

Reversed Phase Column

Abstract We used paired-ion high-performance liquid chromatography to determine the 4-nitrophenol content of 4-nitrophenyl phosphate, a substrate for alkaline phosphatase analysis. This was done on a reversed-phase column with a mobile phase of methanol/water, 45/55 by vol, containing 3 ml of tetrabutylammonium phosphate reagent per 200 ml of solvent. At a flow rate of 1 ml/min, 4-nitrophenol was eluted at 9 min and monitored at 404 nm; 4-nitrophenyl phosphate was eluted at 5 min and could be monitored at 311 nm. Samples of 4-nitrophenyl phosphate obtained from several sources contained 0.3 to 7.8 mole of 4-nitrophenol per mole of 4-nitrophenyl phosphate.

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Simultaneous Determination of Preservatives (Methyl Paraben and Propyl Paraben) in Sucralfate Suspension Using High Performance Liquid Chromatography

E-Journal of Chemistry ◽

10.1155/2011/360431 ◽

2011 ◽

Vol 8 (1) ◽

pp. 340-346 ◽

Cited By ~ 5

Author(s):

Rajesh M. Kashid ◽

Santosh G. Singh ◽

Shrawan Singh

Keyword(s):

High Performance Liquid Chromatography ◽

Simultaneous Determination ◽

Mobile Phase ◽

High Performance ◽

Reversed Phase ◽

Hplc Method ◽

Reversed Phase Hplc ◽

Methyl Paraben ◽

Propyl Paraben

A reversed phase HPLC method that allows the separation and simultaneous determination of the preservatives methyl paraben (M.P.) and propyl paraben (P.P.) is described. The separations were effected by using an initial mobile phase of water: acetonitrile (50:50) on Inertsil C18 to elute P.P. and M.P. The detector wavelength was set at 205 nm. Under these conditions, separation of the two components was achieved in less than 10 min. Analytical characteristics of the separation such as precision, specificity, linear range and reproducibility were evaluated. The developed method was applied for the determination of preservative M.P. and P.P. at concentration of 0.01 mg/mL and 0.1 mg/mL respectively. The method was successfully used for determining both compounds in sucralfate suspension.

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