High Performance Liquid Chromatography Kangfu Capsules matrine, matrine oxide
【Abstract】 Objective: To establish Kang Capsule Women method for the determination. Methods: The high-performance
liquid chromatography. Column: Agilent Technologies ZORBAX
Extend-C18 4.6 × 250mm, 5μm, mobile phase: acetonitrile -
methanol - phosphate buffer (PH6.8) (16: 16:68), flow rate: 1.0ml * min-1 The detection wavelength: 220nm, column temperature: 30 ?. Results: matrine at 0.02 ~ 0.40mg * ml-1
range of a good linear relationship (r = 0.9995); The average recovery was 98.2%, RSD = 1.4% (n = 6); oxidation of matrine at 0.01 ~ 0.20mg * ml-1 range of a good linear relationship (r = 0.9997); The average recovery was 97.6%, RSD = 2.2% (n = 6). Conclusion: This method is simple, accurate and reliable, suitable for women Kang Capsule quality control.
Key words high-performance liquid chromatography Kangfu Capsules Determination of matrine oxymatrine
Determination of Matrine Oxgmatrine content in Kangfuling
capsules by HPLC
【Abstract objective】 To set up a method for quality
control of Fuyankang tablets. METHOD: HPLC method was developed to quantitative determination. The separation was performed on Agilent Technologies ZORBAX Extend-C18 4.6 ×
250mm, 5μm.The mobile phase was acetonitrile-methanol -
phosphate buffer (pH = 6.8) (16:16:68). The flow rate was 1.0ml * min-1. The UV detection wavelength was 220nm. The column temperature was 30 ?. RESULTS: The linear range of
Matrine and Oxgmatrine were at 0.02 ~ 0.40 mg * ml-1 (r =
0.9995) and 0.01 ~ 0.20 mg * ml-1 (r = 0.9997) respectively,
the average recoveries (n = 6) were 98.2% and 97.6% with RSD 1.4% and 2.2%. CONCLUSION The method is simple and accurate, it can be used for quality control of Kangfuling capsules.
【Key words】 HPLC; Kangfuling capsules; Matrine;
Kang Ling Capsule women as "the State Drug Administration Standard (Trial)" collection contains varieties, from Sophora flavescens, bar perfoliatum, Treats, Motherwort, Millettia,
safflower gentian, soil Fuling, Chinese angelica, etc. 8 kinds of traditional Chinese medicine composition. With a heat dampness, promoting blood circulation to regulate menstruation only with the efficacy of commonly used traditional Chinese medicine for gynecological . Kushen containing matrine, matrine oxide and other ingredients, we use high-performance
liquid chromatography preparation of matrine, matrine oxide content, the method is simple, fast, accurate and reliable, and can be used for health Women's Capsules quality control.
An instrument and reagent
1.1 Instrument LC-2010A high-performance liquid
chromatography, CLASS-VP chromatography workstation.
1.2 Test drug matrine reference substance (Lot No. :100078-200414), oxymatrine reference substance (Lot
No. :110780-200004), by China's pharmaceutical and biological products to provide for the determination of use; Kang Ling Capsule Women for the commercially available samples (Guizhou and Ren Tang Pharmaceutical Co., Ltd., Spec: 0.4g * grain -1,
batch number: 20061108,20061202,20061216). Acetonitrile, methanol for HPLC pure water to ultrapure water, other reagents were analytical pure.
2 Methods and Results
2.1 Chromatographic conditions and system suitability
test Column: Agilent Technologies ZORBAX Extend-C18 4.6 ×
250mm, 5μm, mobile phase: acetonitrile - methanol - phosphate
buffer (PH6.8) (16:16:68) The detection wavelength: 220nm,
flow rate: 1.0ml/min, column temperature: 30 ?.
2.2 Solution preparation
2.2.1 Preparation of reference substance solution precision vacuum drying, said phosphorus pentoxide learn more than 12 hours of matrine reference substance and oxymatrine the amount of reference substance, and each made of methanol, each containing 0.4mg per 1ml of the control Commodities reserve liquid; then take precise amount of liquid reserves of matrine reference substance and oxymatrine reference substance reserve the amount of liquid, add methanol produced per 1ml of matrine containing 0.2mg, Oxymatrine 0.1mg mixed solution, As a reference substance solution.
2.2.2 Preparation of sample solution taken Kang Capsule 10 Women's contents, and research fine, take about 1.0g, said precision set, and placing a plug conical flask, add concentrated ammonia solution 2ml so moist, coupled with trichloro A Alkyl (CHCl3) 30ml, ultrasonic treatment (power 250W, frequency 33KH2) 20 minutes, filtration, take filtrate, and then chloroform washing residues, containers and filters four times, each 5ml, filtration, combined filtrate , home on the water bath evaporated, residue was dissolved in methanol and transferred to a 25ml Liang Ping, add methanol to the scale, shake, filtration, take added filtrate that was.
2.2.3 Preparation of negative control solution except to
take the rest of Sophora prescription medicines other than the one-tenth of the amount, according to the rule of law into the film, and then sample preparation methods, made of negative control solution.
2.3 Specificity Test
Precision drawing sample solution, respectively, negative control solution and the reference substance solution of 10μ
l, into the liquid chromatograph, record chromatograms (Figure 1). Can be seen from Figure 1 for the test materials chromatography, chromatography with the reference substance for the corresponding position, have the same retention time (1 Oxymatrine: 6.866min; 2 matrine: 15.422min) chromatographic peak negative test without interference, to prove feasibility of this Act.
2.4 The linear range of the study
Precision drawing reference substance reserve the amount of liquid, respectively, increases the concentration of methanol were dubbed matrine: 0.02,0.05,0.10,0.20,0.30,0.40 mg * ml-1, Oxymatrine: 0.01,0.03,0.05,0.10 , 0.15,0.20 mg * ml-1
solution, shaken, filtration, precision drawing continued
filtrate of 10μl, into the liquid chromatograph, record the peak area. To peak area (A) for the vertical axis, and its concentration (C) as the abscissa, linear regression, matrine regression equation: A = 4.34 × 105C +3.87 × 102, r =
0.9995; oxymatrine regression equation To: A = 1.21 × 104C
+8.07 × 102, r = 0.9997. The results showed that matrine at 0.02 ~ 0.04mg * ml-1 within the peak area is linear with
concentration; oxymatrine in 0.01 ~ 0.20mg * ml-1 within the
peak area is linear with the concentration of relationship.
To take samples (Guizhou and Ren Tang Pharmaceutical Co., Ltd., batch number: 20,061,108) by "2.2 Preparation of sample solution" approach to prepare sample solution, repeat sampling five times sample volume 10μl, obtained under the above-
mentioned chromatographic conditions were Sophora alkali peak area RSD was 0.9%, peak area RSD Oxymatrine was 1.2%, indicating good precision. Reposted elsewhere in the paper for free download http://
2.6 Stability Test
To take samples (Guizhou and Ren Tang Pharmaceutical Co., Ltd., batch number: 20,061,108) solution, at 0,2,4,8,24 h were measured. The results showed that the sample solution was stable within 24h, matrine peak area
The RSD is 1.1%, peak area RSD Oxymatrine 0.8%.
2.7 Repeatability test
To take samples (batch number: 20,061,108) 6 copies, respectively, according to "2.2 Preparation of sample solution" sample solution was prepared to carry out the determination of matrine content obtained RSD was 0.7% RSD
oxymatrine content of 1.6 %, indicating good reproducibility.
2.8 The average recovery test
Precision said the levels are known to take samples (Guizhou and Ren Tang Pharmaceutical Co., Ltd., batch number: 20,061,108, matrine concentration 3.01mg * g-1, oxymatrine
content of 2.42mg * g-1, average installed capacity 0.3916g * grain -1) amount, a total of six copies, respectively, cone-
shaped bottle home with Sierra Leone, respectively, by adding matrine precision reference substance solution (0.2001mg * ml-
1), reserve liquid Oxymatrine reference substance (0.1000mg * ml-1) the amount, to shake off methanol, according to "2.2 Preparation of sample solution" approach to operations, may recovery test solution, according to the measurement results
in Table 1.
2.9 Determination of Sample
Precision for the test items were drawn with the reference substance solution, the solution of 10μl, into the
liquid chromatograph, three batches of samples determination of matrine, Oxymatrine peak area by external standard method to calculate its concentration (n = 5), The results in Table 2. According to the results of three batches of samples measured is tentatively scheduled for each piece of matrine Kang Capsule content of women's quality control limit of
quantification of 1.8mg, oxymatrine quality control limit of quantification for the content of 0.5mg. Table 1 matrine, Oxymatrine recoveries test in Table 2 Determination results of three batches of samples
3.1 The mobile phase selection I have chosen three kinds of mobile phase: ? acetonitrile -0.1% phosphoric acid (20:80)
(PH value triethylamine adjusted to 8.0) ; ? acetonitrile
- methanol - phosphate buffer (PH6.8) - triethylamine (18:
18:70:0.1) ; ? acetonitrile - methanol - phosphate buffer
(PH6.8) (16: 16:68). After repeated tests showed that the first two peak shape derived from a wide mobile phase containing impurities, while the use of ? acetonitrile -
methanol - phosphate buffer (PH6.8) as mobile phase and the
sample obtained a good peak shape, interference fewer
components, was chosen as the determination of the mobile phase.
3.2 The author compares the set of solvent dissolved in methanol for the test chemicals and dissolved in mobile phase
for the test materials, the results of using methanol as the solvent separation is good, and the retention time is short.
3.3 by three batches of samples of matrine, the determination of Oxymatrine results show that the highest level of 1.38mg * matrine grains -1, a minimum of 1.18mg *
tablets -1; oxymatrine a maximum content of 1.08 mg * grain -
1, a minimum of 0.94mg * grain -1. Comprehensive consideration
to determine limits for each piece of matrine containing not less than 1.8mg, with Oxymatrine not less than 0.5mg.
This method is accurate, method is simple, reproducible, and recoveries were satisfactory, you can effectively control the product quality.
 "the State Drug Administration Standard (Trial)" [S].
 Chinese Pharmacopoeia [S]. 2005,497 ~ 498. Reposted elsewhere in the paper for free download http://