Separation and determination of betaine in an oriental medicine by liquid chromatography

Kikuchi, Nobu; Matsuno, Kyoko; Miki, Taihei

doi:10.1016/0003-2670(93)85241-b

Cited by 10 publications

(7 citation statements)

References 21 publications

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“…Most procedures have used organic solvents for the extraction of betaine from organic materials 13. 14 However, they have extracted betaine without the use of heat. Gorham15 used hot methanol to extract betaine from plant samples.…”

Section: Resultsmentioning

confidence: 99%

“…One of the main problems encountered with other assay methods14, 18 for betaine was the purification procedure. When used on our test substance, alfalfa, these procedures were unable to remove a number of substances, which then co‐eluted with betaine.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to this, 1 H NMR spectroscopy equipment is not part of many nutrition laboratories. Other methods14, 17 developed so far have not been used to analyse betaine in different feed ingredients.…”

Section: Resultsmentioning

confidence: 99%

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Determination of the betaine content of feed ingredients using high‐performance liquid chromatography

et al. 2002

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A series of studies was conducted to establish a methodology for the accurate and efficient determination of betaine in different feed ingredients. The final methodology involves an extraction step in which the feed sample is heated for 3 h in a methanolic KOH solution using a Goldfisch apparatus. Impurities are removed by the addition of activated charcoal and concentrated (36%) HCl. After centrifugation the extractant is passed through a strong cation exchange resin (Dowex 50W-X12, H þ ). The betaine retained in the column is eluted with 1.5 N HCl. A 2 ml aliquot of the elute is air dried and reconstituted with 1 ml of deionised water. HPLC separation with a cation exchange column (Partisil SCX-10) is used for the separation of betaine from other compounds. The mobile phase is kept constant at 50 mM KH 2 PO 4 in water, and eluted compounds are detected by UV absorbance (200 nm). The flow rate is maintained at 1.5 ml min À1 . This assay is very accurate over the range of betaine concentrations from 15 to 650 mg ml À1 , with a lower detection limit in feeds of approximately 500 mg g À1 when 4 g of sample is extracted. Recovery assays done with standard betaine hydrochloride and hard red wheat resulted in a consistent recovery of 80%. Betaine content was quantified in several feed ingredients, including alfalfa (1.77 mg kg À1 ), wheat (3.96 mg kg À1 ), wheat middlings (4.98 mg kg À1 ) and poultry meal (0.77 mg kg À1 ). Betaine in corn and soybean meal was not detectable by this method, even when 16 g of sample was used (<125 mg kg À1 ). Betaine present in several feed ingredients should influence choline supplementation to animal feeds and may have implications for human health.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Determination of the betaine content of feed ingredients using high‐performance liquid chromatography

et al. 2002

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“…Glycine betaine was determined by HPLC, using a cationic exchange column (Dionex Hypersil SCX, 5 µm, 250 × 4.6 mm) at 30°C in isocratic conditions at a flow rate of 1 ml min −1 . The eluent phase was: sodium phosphate buffer 0.05 m , pH 3.7 (95%), methanol (5%) (Kikuchi et al ., 1993). Aliquots of the water extract (40 µl) were injected.…”

Section: Methodsmentioning

confidence: 99%

Free amino acids and glycine betaine in leaf osmoregulation of spinach responding to increasing salt stress

et al. 2003

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Summary• The aim of the paper was to determine nitrogen compounds contributing to leaf cell osmoregulation of spinach ( Spinacia oleracea ) submitted to increasing salt stress.• Sodium, free amino acids and glycine betaine contents were determined in the last fully expanded leaf of plants stressed by daily irrigation with saline water (0.17 M NaCl).• After 20 d of treatment, when Na + content was c . 55 umol g − 1 f. wt above the control, and the reduction of stomatal conductance lowered photosynthesis to c . 60% of the control, the free amino acids of the leaves, especially glycine and serine, strongly increased. Proline and glycine betaine also increased significantly. After 27 d of treatment, when the Na + content was c . 100 umol g − 1 f. wt above the control and photosynthesis was 33% of the control, the free amino acid content, especially glycine and serine, declined. Gycine betaine, but not proline, increased further.• Glycine betaine comprised c . 15% of the overall nitrogen osmolytes at mild saltstress, but represented 55% of the total, when the stress became more severe. The increase of glycine betaine balanced the decline in free amino acids, mainly replacing glycine and serine (the precursors of glycine betaine) in the osmotic adjustment of the cells. Photorespiration, which increased during salt stress, was also suggested to have a role in supplying metabolites to produce compatible osmolytes.

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“…1 Several analytical techniques have been reported for the quantitative determination of Be.Cl in pure solutions and pharmaceutical preparations. Among these are HPLC, [2][3][4][5][6][7] TLC, 8 capillary electrophoresis, determination of ions in solution with high selectivity. Most ISE's are of low-cost, their use and maintenance being very simple, assay procedures involving such electrodes are generally simple and fast.…”

Section: Introductionmentioning

confidence: 99%

Potentiometric Batch and Flow Injection Analysis of Betaine Hydrochloride

2007

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Novel PVC membrane electrodes for the determination of betaine ion based on the formation of betaine-tetraphenylborate (Be-TPB) and betaine-phosphotungstate (Be-PT) ion-exchangers as electroactive materials are described. The sensors show a fast, stable, near Nernstian response for 6.92 x 10(-6) to 7.94 x 10(-3) M and 1.0 x 10(-4) to 1.0 x 10(-2) M betaine hydrochloride (Be.Cl) in case of Be-TPB electrode applying batch and flow injection analysis (FIA), respectively, and 2.95 x 10(-5) to 2.26 x 10(-3) M and 3.16 x 10(-5) to 1.0 x 10(-2) M in case of Be-PT electrode for batch and FIA electrodes, respectively, at 25 degrees C over the pH range of 3.5-10 with a cationic slope of 60.2 and 59.1 mV decade(-1) and a fast potential response of < or =15 s. The lower detection limits are 7.94 x 10(-6) and 3.18 x 10(-5) M Be.Cl for Be-TPB and Be-PT electrodes, respectively. Selectivity coefficient data for some common inorganic cations, sugars, amino acids and the components other than betaine, of the mixed drug investigated show negligible interference. The electrodes have been applied to the direct potentiometric determination of betaine hydrochloride in water and in a pharmaceutical preparation under batch and FIA conditions. Potentiometric titrations of Be.Cl with NaTPB and PTA as titrants were monitored with the developed betaine electrodes as an end point indicator electrode. The determination of Be.Cl shows an average recovery of 100.8% with mean relative standard deviation of 0.61%. The effect of temperature on the electrodes was also studied.

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Separation and determination of betaine in an oriental medicine by liquid chromatography

Cited by 10 publications

References 21 publications

Determination of the betaine content of feed ingredients using high‐performance liquid chromatography

Determination of the betaine content of feed ingredients using high‐performance liquid chromatography

Free amino acids and glycine betaine in leaf osmoregulation of spinach responding to increasing salt stress

Potentiometric Batch and Flow Injection Analysis of Betaine Hydrochloride

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