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Determination method of contents of saccharate and saccharic acid 1,4-lactone in Liuwei Dihuang preparations

A technology of glucaric acid and Liuwei Dihuang, which is applied in the field of glucaric acid salt and glucaric acid 1, to achieve a wide range of pharmacological effects and to clarify the effect of the target

Active Publication Date: 2014-12-31
瑞金市工业投资发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although gluconate is found in fruits such as apples and grapes and a small amount of vegetables, so far, there is no report on their quantitative determination by HPLC-HILIC-UV / ELSD and HPLC-MS / MS

Method used

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  • Determination method of contents of saccharate and saccharic acid 1,4-lactone in Liuwei Dihuang preparations
  • Determination method of contents of saccharate and saccharic acid 1,4-lactone in Liuwei Dihuang preparations
  • Determination method of contents of saccharate and saccharic acid 1,4-lactone in Liuwei Dihuang preparations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Determination of Total Gluconate in Liuwei Dihuang Pills by HPLC-HILIC-UV / ELSD

[0030] 1) Glucaric acid 1,4 lactone is converted into glucaric acid salt

[0031] with different concentrations of Na 2 CO 3The solution was dissolved to prepare glucaric acid 1,4-lactone standard substance, and analyzed by HPLC-HILIC-UV to determine the conditions for complete conversion of glucaric acid 1,4-lactone into glucaric acid salt. The results showed 2.0% Na 2 CO 3 The solution can completely convert glucaric acid 1,4 lactone into glucaric acid salt.

[0032] 2) Content determination

[0033] Take Liuwei Dihuang Wan (Beijing Tongrentang, concentrated pill), crush it, weigh 500mg accurately, add 4.0Ml 2% Na 2 CO 3 The solution was sonicated for 20 minutes, centrifuged, precipitated and extracted once with an equal volume of the above solution, centrifuged, and the supernatant was combined; extracted with a C18 SPE small column, collected the eluted part of 5% methanol, dr...

Embodiment 2

[0037] Determination of Glucose 1.4-Lactone in Liuwei Dihuang Pills by UPLC-MS / MS

[0038] Take Liuwei Dihuang Pills (Beijing Tongrentang, concentrated pills), crush them, weigh 500mg accurately, add 4.0 mL of 60% methanol solution, ultrasonicate for 20 minutes, centrifuge at 5,000 rpm for 10 minutes to take the supernatant, add 4.0 mL of 60% methanol solution to the residue, and ultrasonically After 20 minutes, centrifuge at 5,000 rpm for 10 minutes to take the supernatant, combine the supernatants, centrifuge at 14,000 rpm, and then dilute appropriately for UPLC-MS / MS analysis.

[0039] Chromatographic conditions: Waters BEH C18 column, length 50mm, inner diameter 1.0 mm, particle size 1.7um, column temperature: 40°C. Mobile phase: Phase A is 10mM formic acid, phase B is acetonitrile; flow rate is 0.4mL / min. Mass spectrometry conditions: Electrospray negative ion source (ESI-), voltage 2000V, source temp 100°C, cone voltage 15V, desolvation temperature 300°C, cone gas and d...

Embodiment 3

[0041] Simultaneous Determination of Gluconate and Glucose 1.4-Lactone in Liuwei Dihuang Pills by HPLC-LILIC-UV

[0042] Take Liuwei Dihuang Pills (Beijing Tongrentang, concentrated pills), crush them, weigh 500mg accurately, add 4.0 mL of 60% methanol solution, ultrasonicate for 20 minutes, centrifuge at 5,000 rpm for 10 minutes to take the supernatant, add 4.0 mL of 60% methanol solution to the residue, and ultrasonically After 20 minutes, centrifuge at 5,000 rpm for 10 minutes to take the supernatant, combine the supernatants, centrifuge at 14,000 rpm, and then dilute them appropriately for HPLC-LILIC-UV analysis.

[0043] Chromatographic conditions for HPLC-HILIC-UV method: the chromatographic column is ZIC-HILIC with a length of 250mm, an inner diameter of 4.6mm, and a particle size of 5um. Using acetonitrile (A)-5mM ammonium dihydrogen phosphate + 1.0% phosphoric acid (B) as the mobile phase, the mobile phase gradient elution program: 0-3min A phase 83%, 3-15min A phas...

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Abstract

The invention belongs to the field of pharmaceutical analysis, and relates to a determination method of contents of saccharate and saccharic acid 1,4-lactone, two newly-discovered pharmacologically-active components, in Chinese patent medicine Liuwei Dihuang series preparations. According to the invention, saccharic acid (saccharate) and (or) saccharic acid 1,4-lactone in Liuwei Dihuang preparations are extracted by a Na2CO3 solution or a 60% methanol solution. The two compounds are analyzed quantitatively by separating detection technologies such as hydrophilic chromatography HPLC-HILIC-UV / ELSD / MS, and the like. The invention establishes the determination method of contents of saccharate and its lactone in Liuwei Dihuang preparations for the first time, and provides a new thinking and method for quality control of Liuwei Dihuang preparations.

Description

Technical field [0001] The invention involves the field of drug analysis, which specializes in the measurement method of glucose diite and glucose dilate 1,4 glycodic acid in the six -flavors of the six flavors. Background technique [0002] Liuwei Dihuang Wan is compatible with 6 flavors of Chinese medicine such as cooked Dihuang, dogwood, yam, Zexie, peony skin, and Poria. It has a clinical application history of hundreds of years and is a classic famous prescription for traditional Chinese medicine.With the continuous deepening of clinical research, its attendance has also expanded from the original pediatric drugs to inside, outside, women, children, and facial features. It has more than 40 types of diseases.EssenceDue to the exact effect of the medicine, there are almost no adverse reactions, and the low price and other factors are favored by the public.At present, there are about hundreds of manufacturers in China to produce Liuwei Dihuang products, mainly soup agents, pill...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 谢宝刚张宁宁姚艳琴
Owner 瑞金市工业投资发展有限公司
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