Retention behaviour of cardiac steroids using cyclodextrin in the mobile phase in high-performance liquid chromatography

Shimada, Kazutake; Oe, Tomoyuki; Hirose, Y.; Komine, Yoshihiro

doi:10.1016/0021-9673(89)90036-8

Cited by 8 publications

(6 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have reported that the inclusion HPLC using CD as a mobile phase additive was of great advantage in the separation of isomeric steroids. 13,14 The effect of Me-β-CD and γ -CD in the separation of the two epimers were examined, and the obtained chromatograms are shown in Fig. 2.…”

Section: Identification Of 3-epi-2425(oh)2d3 In Rat Plasmamentioning

confidence: 99%

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

2000

Self Cite

View full text Add to dashboard Cite

(24R)-24,25-Dihydroxyvitamin D3 [24,25(OH)2D3] is one of the major metabolites of vitamin D3, which causes a marked increase in bone volume 1 and mechanical strength 2 in animals without hypercalcemia at pharmacological doses ( Fig. 1). Based on these criteria, the metabolite is expected to be a new antiosteoporosis medicine and much interest is focused on the metabolism of 24,25(OH)2D3. 24,25(OH)2D3 is reported to be oxidized on its side chain at the C-23 or 24 position by vitamin D 24-hydroxylase to give metabolites such as (23S)-23, 25-dihydroxy-24-oxovitamin D3. 3 In previous studies, we found that in rats dosed with 24,25(OH)D3 per os, a large amount of 3-epi-24,25(OH)2D3 24-glucuronide (24G) was excreted into the bile together with 24,25(OH) 2D3-3G, -24G and -3-sulfate. 4,5 The epimerization of the 3-hydroxy group was also observed in the in vitro 6,7 and in vivo 8 metabolism of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3]. These findings indicate that the C-3 epimerization pathway plays an important role as does the side-chain oxidation in the vitamin D metabolism. However, the existence of the free form of the 3-epimer of 24,25(OH)2D3 in biological fluids and the epimerization mechanism have not been clarified. Studies on the 3-epi-metabolites are important to develop new vitamin D medicines. For example, if 3-epi-24,25(OH)2D3 is not only excreted into the bile as a glucuronide but also circulates in the blood, some contribution of the 3-epi-form to the bone formation caused by the administration of 24,25(OH)2D3 must be considered.In the present paper, we initially identified 3-epi-24,25(OH)2D3 in the blood of rats dosed with 24,25(OH)2D3 using inclusion high-performance liquid chromatography (HPLC), liquid chromatography (LC)/mass spectrometry (MS) and gas chromatography (GC)/MS. Second, in vitro experiments were performed to investigate the C-3 epimerization mechanism of 24,25(OH)2D3 using the deuterium-labeled material. . 4-Phenyl-1,2,4-triazoline-3,5-dione (PTAD) was synthesized from 4-phenylurazole (Nacalai Tesque, Kyoto, Japan) and purified by sublimation. 9 Isolute C18 (EC) cartridges (500 mg; International Sorbent Tech., Ltd., Hengoed, UK) were obtained from Uniflex (Tokyo). Heptakis-(2,6-di-O-methyl)-β-cyclodextrin (Me-β-CD) and γ -CD were donated by Kao Co. (Tokyo) and Nihon Shokuhin Kako Co. (Tokyo), respectively. All other reagents and materials were of analytical grade. Experimental Materials and reagents HPLC, LC/MS and GC/MSHPLC was performed using a Hitachi L-7110 chromatograph (Tokyo) equipped with a Shimadzu SPD-10A UV (265 nm) detector (Kyoto). LC/MS was performed using a Finnigan MAT LCQ (San Jose, CA, USA) connected to a JASCO PU-980 (Tokyo) chromatograph, and atmospheric pressure chemical ionization (APCI) was used in the positive-ion mode. The heated capillary temperature was set at 150˚C and 225˚C for analyses of the intact vitamin D compounds and the PTADadducts, respectively. The sheath gas flow rate was set at 80 units with a vaporizer temperature of 350˚C. The source current, th...

show abstract

Section: Identification Of 3-epi-2425(oh)2d3 In Rat Plasmamentioning

confidence: 99%

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

2000

Self Cite

View full text Add to dashboard Cite

show abstract

“…Mobile phase additives are known means by which the separation on reversed phase columns might be improved. [12][13][14] Additives such as urea (0.2 g=100 mL), ammonium acetate (0.2 g=100 mL), triethylamine (0.5 mL=100 mL), and hydroxy propyl B-cyclodextrin were examined. Overall, there were no significant changes in relative retention of the test compounds, i.e., the best result obtained was quite similar to the optimum obtained using the C 18 column with 75% methanol without additives (see chromatogram in Figure 3).…”

Section: Resultsmentioning

confidence: 99%

Development and Validation of a HPLC Method for Determination of Potential Residual Cortisone Compounds in Timolol Maleate Eye Drops

Hamdan

Qurani

2008

Journal of Liquid Chromatography & Related Technologies

View full text Add to dashboard Cite

One way of ensuring effective cleaning in the pharmaceutical industry is to analyze the batch of a product for a potential carry over contamination from a previously manufactured one. The potential cross contamination of timolol maleate (TM) eye drops with cortisone compounds, deserves special consideration, due to the well known deleterious effect of cortisones on glaucoma. Therefore, it was the objective of this study to develop and validate a HPLC method for the detection and quantitative determination of cortisones in TM eye drop preparations. The chromatographic behaviors of the five cortisones that were most likely to be present as contaminants in TM eye drops were characterized on different HPLC stationary phases (normal silica, C 18 , C 8 , and CN). Mobile phase and buffer constituents were further optimized on the best stationary phase material (C 18 ). The final recommended mobile phase consisted of a mixture of THF:methanol:0.01M phosphate buffet at ratios of 15:25:60 and containing 0.01 M camphore sulfonic acid (pH adjusted to 4.2). The method was validated in light of ICH guidelines and applied to commercial samples with satisfactory results.

show abstract

“…These results prompted us to use CD as the mobile phase additive, which has been proved to be a great advantage in the separation of isomeric steroids. 7,8 Me-β-CD and γ-CD were used based on their solubility in organic solvents 9 and inclusion effect for steroids. The combination of organic modifiers and CDs were systematically examined, and two mobile phase systems shown in Figures 2a and b were found to give a good separation of 2, 3, and 5.…”

Section: Derivatization With Ptadmentioning

confidence: 99%

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION OF DIHYDROXYLATED VITAMIN D₃METABOLITES USING MOBILE PHASE CONTAINING CYCLODEXTRIN

Higashi

Ogasawara

Shimada

2000

Journal of Liquid Chromatography & Related Technologies

View full text Add to dashboard Cite

The separation of side chain dihydroxylated vitamin D 3 metabolites by inclusion high performance liquid chromatography using cyclodextrin (CD) as the mobile phase additive was examined. 24,25-Dihydroxyvitamin D 3 and its 3-epimer were clearly separated by the addition of γ-CD to the mobile phase using acetonitrile as the organic modifier. The separation of the 25-epimers of 25,26-dihydroxyvitamin D 3 has been done, to a certain extent, with the mobile phase using methanol and dimethylated β-CD as the organic modifier and the additive, respectively. These separation methods were applied to the detection of the metabolites in the plasma of the rat administered 25-hydroxyvitamin D 3 . These metabolites were also identified by liquid chromatography/tandem mass spectrometry after derivatization.

show abstract

Retention behaviour of cardiac steroids using cyclodextrin in the mobile phase in high-performance liquid chromatography

Cited by 8 publications

References 18 publications

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

Development and Validation of a HPLC Method for Determination of Potential Residual Cortisone Compounds in Timolol Maleate Eye Drops

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION OF DIHYDROXYLATED VITAMIN D₃METABOLITES USING MOBILE PHASE CONTAINING CYCLODEXTRIN

Contact Info

Product

Resources

About

Retention behaviour of cardiac steroids using cyclodextrin in the mobile phase in high-performance liquid chromatography

Cited by 8 publications

References 18 publications

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

Investigation of C-3 Epimerization Mechanism of 24,25-Dihydroxyvitamin D3 in Rat Using Liquid Chromatography/Mass Spectrometry

Development and Validation of a HPLC Method for Determination of Potential Residual Cortisone Compounds in Timolol Maleate Eye Drops

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION OF DIHYDROXYLATED VITAMIN D3METABOLITES USING MOBILE PHASE CONTAINING CYCLODEXTRIN

Contact Info

Product

Resources

About

HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION OF DIHYDROXYLATED VITAMIN D₃METABOLITES USING MOBILE PHASE CONTAINING CYCLODEXTRIN