High Performance Liquid Chromatography Xiaotangling capsule content of glibenclamide
【Abstract】 Objective To investigate the high-performance
liquid chromatography (HPLC) Determination of Xiaotangling capsule content of glibenclamide. Methods HP ODS-Hypersil
column (4.6 mm × 200 mm, 5μm), column temperature: 20 ?,
mobile phase was methanol -0.005 mol / L amine dihydrogen
phosphate (pH3.5, 70:30), flow rate 0.8 ml / min, detection wavelength 300 nm. Results of glibenclamide in the context of
0.141 ~ 2.25μg was a good linear relationship (r = 0.999 9, n = 5), the average recovery was 99.18%, RSD of 1.72% (n = 3). Conclusion High Performance Liquid Chromatography glibenclamide simple, fast, sensitive and can be used as Xiaotangling capsules determination method.
Key words Xiaotangling Capsules Glibenclamide high-
performance liquid chromatography of berberine hydrochloride
Determination of Glibenclamide in XiaoTangLing Capsules by HPLC
CUI Zhe, GUO Jian peng
1.The Hospital of Yanbian University, Yanji, Jilin 133000, China;
2. Pharmacy College of Yanbian University, Yanji, Jilin 133000, China
AbstractObjectiveTo research the methods of determination of Glibenclamide content in XiaoTangLing capsules by HPLC.
MethodsHP ODS-Hypersil chromatographic column (4.6 mm × 200
mm, 5 μm) was used to determine the content of Glibenclamide under the temperature of 20 ?. Mobile phase: The ratio of
methanol and 0.005mol / L phosphoric acid dihydroamine was 70:30 under the pH3.5. Flow rate: 0.8 ml / min. The wavelength of detection: 300 nm. ResultsIn the range from 0.141μg to
2.25μg, Glibenclamide appeared fine linear correlation (r = 0.999 9, n = 5). The average recovery was 99.18%, RSD = 1.72% (n = 3). ConclusionHPLC method has some advantages, such as convenient, swift, sensitive, so HPLC can be used as a
suitable method for determination of XiaoTangLing capsule.
Key wordsXiaoTangLing capsule; Glibenclamide; HPLC
Xiaotangling capsule containing relevant in the "ministerial standard" (WS3-B-2026), by the ginseng,
berberine, astragalus and other 11 Chinese herbal medicines out of the urea composition of Gag, with Qi Yin, clearing away heat Xiehuo, kidney the effect of reduction of urine for the treatment of diabetes. Fang Glibenclamide is a widely used clinical hypoglycemic class of synthetic drugs, but also has a certain toxicity and adverse reactions, it is necessary for accurate determination of its content. In order to improve the quality of the preparation standards for high-performance
liquid chromatography to be used glibenclamide as a method for
the determination of this product.
An instrument and reagent
Hewlett-Packard HP1100 high performance liquid
chromatograph, HP1100 chromatography workstation; Sartorious CP225-D-type electronic balance (one ten-thousandth, Germany
SatoriUs companies).
Xiaotangling capsule (Pharmacy, Yanbian University, Laboratory of preparation, batch number:
040603,040605,040607), glyburide reference substance, (Lot 040,411 Bio-Technology Co., Ltd. Shanghai Friends of the Si);
methanol (HPLC pure); water re-distilled water; other reagents
were analytical pure.
2 Methods and Results.
2.1 Chromatographic conditions
Column was HP ODS-Hypersil (4.6 mm × 200 mm, 5μm),
column temperature 20 ?, mobile phase was methanol-0.005mol /
L amine dihydrogen phosphate (pH3.5, 70:30), flow rate 0.8 ml / min, detection wavelength was 300 nm.
2.2 Preparation of reference substance solution
Precision Weigh appropriate glibenclamide reference substance, add methanol produced per ml of glibenclamide 225μ
g of the solution, shake, as a reference substance reserve liquid.
2.3 Preparation of solution for the test items
Cancel Sugar Capsule contents, and research fine (over 80 mesh sieve), precision Weigh 0.5 g, set 50 ml Liang Ping, add methanol 40 ml, sonication 30 min (power 250 W, frequency 33 kHz), plus methanol to the scale, shake, filtration, take added filtrate that was.
2.4 Preparation of negative control solution
Take-negative samples (without glibenclamide) 5 g, according to "Preparation for the test product solution" entry method of preparation, that is too negative control solution.
2.5 System suitability test
According to the above-mentioned chromatographic
conditions, to learn all the above-mentioned three kinds of
solution, 10 μl, were injected into liquid chromatograph. Received glibenclamide reference substance for the test
products and negative reference substance chromatogram (see Figure 1 ~ 3) for the test product with the reference substances have a corresponding position in the same chromatographic peak, the negative reference substance in this reservation time in the vicinity without interference.
Calculation of glibenclamide in theoretical plate number is greater than 5000, and separation is greater than 1.5.
2.6 linear relationship between the study
Reference substances were obtained precise amount of
liquid reserves amount dubbed 225,112.5,56.2,28.1,14.1 μg /
ml solution, shake, the precision drawing 10 ml sample. The chromatographic conditions were determined according to the above peak area values to peak area value of the vertical axis (y), in order to glyburide sample volume (μg) for the
abscissa (x), draw the standard curve regression equation was:
y = 446.46x 8.48, r = 0.999 9, glyburide in the range of 0.141 ~ 2.25 μg showed a good linear relationship within. Reposted elsewhere in the paper for free download http://
Detection of the amount of 2.7 to S / N ? 3 when the
sample size for the minimum detectable quantity, the measured minimum detectable glyburide approximately 10 μg.
2.8 Precision Experiment
To take the same reference substance solution, repeated sampling five times, recording peak area and calculating the RSD = 1.07%, showed a good precision of this Act.
2.9 Stability test
To take the same test materials for the solution of 1,2,4,6,8, respectively, into the sample, record peak area and calculating the RSD = 1.77%, that for the test product solution at 8 h in good stability.
2.10 to reproduce the experiment
Number of samples taken in accordance with the "Preparation for the test product solution," under the method, made five copies for the solution of test items to measure and record peak area and calculating the average concentration of glibenclamide to 1.62 mg / g, RSD 1.47 %, indicating good reproducibility of this Law.
2.11 The average recovery experiment
Precision said the nine were taking the same batch of samples has been detected in content, each of about 0.5 g, respectively, by adding an appropriate reference substance glyburide and click "solution for the preparation of test
items" under way to prepare for the test product solution, measured by the formula: The average recovery (%) = [(measured by volume - the original sample volume) / adding amount] ×
100%, calculated recovery. The results in Table 1. Table 1 The average recovery determination results (omitted)
2.12 Sample measurement
Were collected from different batch Xiaotangling capsule, click "solution for the preparation of test items" entry method, preparation for the test product solution, according to the above chromatographic conditions were determined according to the law. The results in Table 2. Table 2 samples
the content of glibenclamide (omitted)
3 Discussion
3.1 Sample pre-treatment Methods
Xiaotangling capsule is a multi-flavored Chinese medicine
and the composition of glibenclamide compound preparation, determination of interfering with big, it is on the extraction method and extraction time were studied. As a result, the study of reflux extraction, ultrasonic extraction, continuous reflux extraction three, using this law to the active ingredient glibenclamide effect the optimal extraction.
3.2 Determination of Wavelength Selection
By UV spectrophotometer scanning, glyburide in the 300 nm wavelength of maximum absorption wavelength was chosen to determine 300 nm.
3.3 The choice of mobile phase [1,2]
Have experimented with more methanol -0.005 mol / L amine
dihydrogen phosphate (pH3.5, 65:35) and methanol -0.005 mol /
L amine dihydrogen phosphate (pH3.5, 70:30) two kinds of mobile phase, the results , methanol -0.005 mol / L amine
dihydrogen phosphate (pH3.5, 70:30) separation is better, the retention time is moderate, so selection of methanol: 0.005 mol / L amine dihydrogen phosphate (70:30) as mobile phase.
【References】
[1] Wu Jing, Yang Qing. HPLC Determination of compound
metformin glyburide content [J]. Anhui Medicine, 2003,7 (3): 223.
[2] Zhang X, Wang sample. Glibenclamide tablets and its HPLC determination of [J]. China's pharmaceutical industry magazine, 1997,28 (1): 33. Reposted elsewhere in the paper for free download http://www. hi138.com