Dhuka wood particles in the papaverine content in RP-HPLC
Authors: Li Huandan, Ji Zhi, Liu Feng, Tian Jia Jia, Zhou Hongbing
[Abstract] Objective RP-paverine Dhuka
content of wood particles. Method uses DIKMA C8 column (250 mm * 4.6 mm, 5μ m), acetonitrile-
as mobile phase, flow rate 1 ml / min, detection a wavelength of 240 nm. The results of
papaverine in 0.019 8 ~ 0.396μ g showed good linear relationship within the framework of (r = 0.999 9), the average recovery was 99.6%, RSD 1.5%. CONCLUSION The method is sensitive, accurate, specific, reproducible and can be used as Dhuka wood particle method for the determination.
[Key words] Dhuka wood grain high-performance liquid chromatography Papaverine
Abstract: Objective To establish a method for determining papaverine in Zukamu Granules.MethodsThe separation was carried out on DIKMA C8 column (250 mm *
4.6 mm, 5 μ m). The mobile phase was carbinol-acetonitrile-1% triethylamine -1%
ammonium acetate (40:20:1:39) with a flow rate of 1.0ml/min.The detection wavelength was set at 240 nm.ResultsPapaverine showed a good linear relationship in the range of
0.0198 ~ 0.396μ g (r = 0.999 9) and the average recovery was 99.6%, RSD was1.5%. ConclusionThe method is sensitive, accurate, specific and repeatable. It can be used for quality control of Zukamu Granules.
Key words: Zukamu Granules; HPLC; Papaverine
By kaempferol, lilies and other opiate shells composed of Medicine Smell extract Dhuka wood particles, with the heat, sweating, Tongqiao efficacy for the common cold cough, fever without sweat, sore throat, stuffy nose, runny nose embolism. Fang opiate
shells containing papaverine, Jiufu easy to addiction, excessive intake can cause irregular heart beats, in severe cases cause death. Articles explore the use of reversed-
phase high performance liquid chromatography determination of papaverine content,
for the control Dhuka provide the basis for the quality of wood particles.
An instrument and reagent
Shimadzu LC-10AT high-performance liquid chromatography (Shimadzu, Japan),
HW-2000 chromatography workstation, C8 column (250mm * 4.6mm, 5μ m) (Dumas Company), ultrasonic cleaning extractor.
Methanol for chromatography pure (Tianjin 4 Friends of the bio-medical
technology development company), water for ultra-pure water, I reagents for the
analysis of alcohol, papaverine hydrochloride reference substance (China Pharmaceutical and Biological Products Institution, Lot :171214-200403).
2.1 Chromatographic conditions
C8 column with methanol - acetonitrile triethylamine -1% -1% ammonium acetate
(40:20:1:39) as mobile phase, flow rate 1 ml / min, detection wavelength was 240 nm.
2.2 Preparation of solution for the test items
To take this product is about 2.5 g, precision that set, and placing 100 ml bottle with cone-shaped plug, add 10 ml of ammonia test solution after wetting alkalization, add chloroform 50 ml, Zhen Yao 5 min, ultrasound 80 min after the shift to the sub-
funnel, the static layer, sub-taking chloroform layer, water layer with 30 ml chloroform for 3 times extraction, combined chloroform solution, water bath Hui dry chloroform, residue was dissolved in methanol and transferred to 10 ml Liang Ping, add methanol to the scale, shaken, into the pre-column with 0.45 μ m microporous membrane filter,
which was tested product solution.
2.3 Preparation of reference substance solution
Take appropriate papaverine hydrochloride, 105 ? drying 2 h, precision that set,
plus methanol made from 39.6 μ g per ml of solution, as a reference substance solution.
2.4 Preparation of negative control solution
Remove Dhuka prescription of opiate shells wood particles, wood particles produced by Dhuka negative fabricated by agents, according to test products for the
preparation method of solution made from the negative control solution.
3.1 Interference Experiment
According to the above-mentioned chromatographic conditions were obtained for
the test product solution, reference substance solution and the negative control solution of 20 μ l, into the chromatograph, record chromatograms, the figure shows a good peak shape of papaverine hydrochloride, and the peak of the sample separation of impurities , negative control solution without interference. The results shown in Figure 1 ~ 3.
3.2 The linear range of the experimental
Respectively, a certain amount of reference substance precision drawing solution diluted to a concentration of 0.99,1.98,2.97,4.95,9.90,14.85,19.80 μ g / ml of standard solution. According to the above chromatographic conditions were injection 20μ l, taking three times the average peak area of papaverine hydrochloride injection volume (μ g) for linear regression, was the regression equation Y = 4563.5X 18009 (r = 0.999 9, n = 7) The results show that the Papaverine hydrochloride in the range of 0.019 8 ~ 0.396μ g a good linear relationship. Reposted elsewhere in the paper for free download http://
3.3 Precision Experiment
Papaverine hydrochloride obtained in control solution, according to the above chromatographic conditions were a continuous injection six times, 20μ l / times, determine the peak area, its RSD 1.44%, The results show that good precision.
3.4 Repeatability test
To take samples of the same batch, according to '2 .2 'method for the parallel preparation of six copies of test materials solution, respectively determination, RSD of 1.7%.
3.5 Stability test
Obtained for the test product solution, according to the above chromatographic conditions, at intervals of 2 h into the sample a time, recording peak area, results showed that the test items for the solution of the basic stability in the 8 h period, RSD
1.0%. The results in Table 1. Table 1 Stability Experiment (abbreviated)
3.6 The average recovery experiment
Precision Weigh a known concentration of sample (batch 060,728) 6 copies, respectively, the control precision by adding papaverine hydrochloride solution (39.6μ
g/ml) 1ml, press '2 .2 'method of preparation for the test items parallel solution, respectively assay results The average recovery was 99.6%, RSD 1.5%. The results in Table 2.
3.7 Determination of Sample
Take five batches of samples, respectively, '2 .2 'method of preparation for the test items parallel solution, respectively assay. The results in Table 3. Table 2 The average recovery results (omitted)
4.1 The choice of mobile phase
Have been inspected methanol - acetonitrile-triethylamine -1% -1% ammonium
acetate (55:5:1:39); methanol - acetonitrile-triethylamine -1% -1% ammonium acetate
(50:10:1:39); of methanol - acetonitrile triethylamine -1% -1% ammonium acetate
(45:15:1:39); methanol - acetonitrile triethylamine -1% -1% ammonium acetate
(40:20:1:39); methanol -- acetonitrile-triethylamine -1% -1% ammonium acetate
(30:30:1:39), the result of methanol - acetonitrile-triethylamine -1% -1% ammonium
acetate (40:20:1:39) as mobile phase separation of the best results. Table 3 Dhuka wood particle content of the sample determination results (omitted)
4.2 The choice of detection wavelength
To take papaverine reference substance solution, at scan measured on a UV
spectrophotometer at 243.5 nm Department has strong absorption at 280.0 and 311.5 nm Department has a weak absorption, also refer to the literature [1 ~ 4], the choice of 240 nm for the The detection wavelength.
4.3 Preparation for the test solution, the choice of goods
Papaverine content of this product in a relatively small, so using liquid-liquid
extraction methods extract enrichment. Basic approach is the use of papaverine was alkaline, after ammonia or ammonia test solution after alkali free, and then the organic
solvent chloroform extraction. Has compared the two alkaline reagents, alkalinized with ammonia was found prone to emulsification phenomenon, not easy to layer, so use of ammonia test solution as the alkaline reagent. Extraction methods have been
compared on the cold soak method (room temperature for 24 h) and ultrasonic extraction method, results showed that the ultrasonic extraction extraction, high efficiency, the results are stable. In the ultrasonic time, compared 20,40,60,80,100 min
extraction rate, the results demonstrate that the time be extended to 60 min, extraction rates will not increase the basic in order to ensure complete extraction, a conservative approach to ultrasonic time determined to be 80 min.
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