37 superfine quality standard of
【Abstract】 Objective To establish a 37 superfine quality standards. Thin Layer Chromatography 37; the use of laser particle detector for detection of the particle size; HPLC determination of 37 saponins in 37 superfine R1, Ginsenoside Rg1, Ginsenoside Rb1 content. The results of TLC spots were clear patterns can be identified with the corresponding 37 spots; 37 superfine powder, the median diameter at 15 μm
below; Notoginsenoside R1 in the range of 0.535 ~ 3.210 μg
good linear relationship (r = 1.000), The average recovery of 103.08%, RSD = 2.80%; ginsenoside Rg1 in the range of 2.225 ~ 13.350 μg good linear relationship (r = 1.000), average recovery was 100. 33%, RSD = 2.29%; ginsenoside Rb1 in the
2.205 ~ 13.230 μg range a good linear relationship (r =
1.000), average recovery was 101.00%, RSD = 1.43%. CONCLUSION The method is simple, accurate, reproducible and can be used for quality control of 37 superfine
Key words 37 ultrafine particle thin-layer chromatography
quality standards for high-performance liquid
Abstract: ObjectiveTo develop a quality standard for super micropowder of notoginseng.Methods Qualitative analysis of micropowder of notoginseng was carried out by TLC. The
particle diameter was detected with laser scattering particle size distribution anaslyzer. An RP - HPLC method was used for
the content determination of the Notoginsensode R1, ginsenoside Rg1, ginsenoside Rb1. ResultsThe chromatographic
spots of micropowder of notoginseng were identified without the interference of negative control. Micropowder of particle average diameter was below 15μm. Notoginsensode R1 had a good
linearity in the range of 0.535 ~ 3.210 μg (r = 1.000), and
the average recovery was 103.08% with RSD of 2.80%.
Ginsenoside Rg1 had a good linearity in the range of 2.225 ~ 13.350 μg (r = 1.000), and the average recovery was 100.33% with RSD of 2.29 %. Ginsenoside Rb1 had a good linearity in the range of 2.205 ~ 13.230 μg (r = 1.000), and the average
recovery was 101.00% with RSD of 1.43%. ConclusionThis method is simple, accurate and repeatable. It can be used to determine ginsenoside Rg1, ginsenoside Rb1and notoginsensode R1 in micropowder of Notoginseng.
Key words: Micropowder of Notoginseng; Particle diameter;
HPLC; TLC; Quality standard
37 (Radix Notoginseng) is a common precious medicinal materials, currently on the market 37 ultrafine powder has become quite common, but 37 superfine quality standard of the study reported in the literature less, the topic
identification using TLC, using laser particle detector pairs 37 superfine particle size detection, and in reference , based on the simultaneous determination of 37 saponins ultrafine powder products, 37 R1, ginsenoside Rg1 and
ginsenoside Rb1 content. The method is simple, accurate, reproducible and can be used for 37 superfine quality control.
An instrument and reagent
HP1100 high performance liquid chromatography, diode array detector, quaternary pump, Sartorius BP211D electronic analytical balance, Acetonitrile, methanol (HPLC grade), water re-distilled water, Ginsenoside Rg1, Ginsenoside Rb1,
Ginsenoside Re, Notoginsenoside R1 reference substance, and 37 control medicines (both Chinese medicine and biological
products available), 37 laboratory-based preparation of
ultrafine powder samples.
2 Methods and Results
2.1 TLC to take this product 0.8 g, add 5 drops of water, stir well, to be water-saturated n-butanol 5 ml, Mesa, Zhen
Yao 10 min, placed 2 h, centrifuged and the supernatant, plus 3 times the amount in order to n-butanol saturated water,
shake, placed so that is divided into layers (if necessary, centrifugation), taking n-butanol layer, evaporated and the
residue add 1 ml of methanol allows the dissolved, as the tested product. Taking Ginsenoside Rg1, Ginsenoside Rb1, Ginsenoside Re, and 37 made of saponin R1 plus 1 ml of methanol containing 0.5 mg of the mixed solution, as a reference substance solution. 37 controls an alternative
medicine 0.8 g, according to test materials for the solution preparation method, in contrast with the legal system, medicine solution. According to thin layer chromatography [ "Chinese Pharmacopoeia" (2005 edition) ? Department of
Appendix ? B] experiment, learn from the above-mentioned
three kinds of solution, the 10 μl, respectively, at the same
point of sodium carboxymethyl cellulose as a binder of silica gel G plate , to chloroform - methanol - water (13:7:2) 10 ?
overnight placed under the lower solution as the mobile phase, to start out, dried, sprayed with 10% sulfuric acid ethanol
solution, heated to a spot at 105 ? color clear view. Test
products for chromatography, in contrast medicinal chromatography corresponding position, substantially the same color spots. Figure 1.
1. 37 mixed reference substance 2. 37 control medicines
3,4,5. 37 ultrafine powder samples
Figure 1 37 superfine TLC diagram (omitted)
Detection of ultrafine particle size 2.2 37 37 ultrafine
powder samples obtained three groups, each consisting of taking 0.07 g, plus 1 ml of ethanol, after wetting, add 50 ml of water, 1 min immediately after the use of ultrasonic laser particle detector for detection . The results show that the samples taken were median diameter less than 15 μm. The
results shown in Table 1 and Figure 2.
Table 1 37 ultrafine particle size test results (omitted)
Figure 2 37 ultrafine particle size detection
2.3 Determination of 37 superfine
2.3.1 Chromatographic conditions for analysis according to Bartlett ODS column (150 mm × 4. 6 mm, 5 μm); detection
wavelength 203 nm; column temperature 20 ?; a flow rate of 1
ml / min, mobile phase A (acetonitrile) and B (water) for gradient elution. The results in Table 2.
Table 2 gradient elution program table (omitted)
2.3.2 Preparation of standard solutions that take precision Ginsenoside Rg1, Ginsenoside Rb1 and Notoginsenoside R1 was prepared by dissolving appropriate amount of methanol containing 37 saponins R1 reference substance 0.107 mg / ml, ginsenoside Rg1 reference substance 0.455 mg / ml and
ginsenoside Rb1 reference substance 0.441 mg / ml mixture of reference substance solution, using 0.45 μm millipore
2.3.3 Preparation for the test product solutions that take 37 superfine precision of about 1 g, precision adding
methanol 50 ml, said that given the weight placed overnight, returning home on a 80 ? water bath, boil 2 h, let cool, make up the weight of methanol , shaking, filtration, take added filtrate, using 0.45 μm microporous membrane filter that is
too. Reposted elsewhere in the paper for free download http://
2.3.4 Determination of precision drawing reference substance, respectively, and test items for the solution of the 10 μl, injection of high-performance liquid
chromatography to measure the value of peak area. By external standard method with peak area, that is to get.
2.3.5 Separation results in the above-mentioned
chromatographic conditions, Ginsenoside Rg1, Ginsenoside Rb1 and Notoginsenoside R1 and coexistence of other chemical
components to achieve a good separation. The results shown in Figure 3.
1. Notoginsenoside R1 2. Ginsenoside Rg1 3. Ginsenoside Rb1
a 37 b 37 ultrafine powder samples of reference substances
Figure 3 37 ultrafine high-performance liquid
chromatography mapping (abbreviated)
2.3.6 Preparation of standard curve obtained standard mixed solution, respectively, sample 5,7,10,15,20,25,30 μl,
with peak area of the vertical axis Y, sample volume (μg) for
the abscissa X, linear regression. The results in Table 3.
2.3.7 Experimental precision for the test products to take the same solution, according to the above chromatographic conditions, a continuous injection six times, measured peak area values of 37 saponins R1, ginsenoside Rb1 and ginsenoside Rg1, RSD were 1.45% , 1.81%, 2.02%.
2.3.8 stability test to take the same sample solution, placed 0, 1, 2, 4, 6, 8 h, according to the above chromatographic conditions and peak area were measured by the
value of 37 saponins R1, ginsenoside Rb1 and ginsenoside Rg1, RSD were 1.68%, 1.81%, 2.43%.
2.3.9 Repeatability precision experiments that take the same batch of 37 samples of six copies of ultrafine powder, according to the above method test samples were determined notoginsenoside R1, ginsenoside Rb1 and ginsenoside Rg1
content of the results of the 37 saponins R1 the average content of 0.82%, RSD was 1.84%; ginsenoside Rb1, the average content of 3.36%, RSD was 2.38%; ginsenoside-Rg1, the average
content of 3.01%, RSD 2.54%.
2.3.10 The average recovery experiment to take samples of
known content in the same batch, and research fine, precision that take six copies, each of approximately 1 g (equivalent to approximately R1 saponins including 37 0.008 1 g, about 0.030 ginsenoside Rg1 8 g and ginsenoside Rb10.030 7 g), according
to the above method of experiment, the samples were determined notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 content of the results of the average recoveries were 103.08%, 100.33%, 101.00% , RSD were 2.80%, 2.29%, 1.43% (n = 6). The
results in Table 4.
Table 3 37 superfine standard curve (omitted)
Table 4 37 superfine recoveries (omitted)
2.3.11 Determination of content of the sample according to the above-mentioned chromatographic conditions and testing methods for the 37 pairs of self-determination of ultrafine
powder samples to measure the results. The results in table 5.
Table 5 Sample assay results (omitted)
Ultrafine grinding technology is to meet the needs of
modernization of Chinese medicine developed a new technique, 37 by the ultra-fine powder, its powder nature of some changes that might be the quality of superfine powder 37 an impact on this end, we have 37 superfine quality standards were studied.
Using TLC, size detection and determination of the method of combining pairs of superfine quality standard of 37 studies, the results of TLC of 37 superfine clear, with the reference substance substantially in the same position relative to the same particle size detection The median
diameter can be seen in the 10.74 μm, in line with USP
standards for determination.
Superfine quality standard of 37 studies, but also for the control of a variety of 37 superfine powder formulations on the market has established quality control standards, which regulate the market.
In TLC, the study of the mobile phase ethyl acetate, chloroform methanol-water (15:40:22:10) and found
that after the commencement of spots in the more casual, non-
round, the use of chloroform methanol-water (13:7:2),
the spots round the whole, a good separation.
In this study, particle size control, not only using the laser particle detector was determined, but also examines the
use of electron microscopic observation of ultrafine particle size, observed that the ultrafine particle size were smaller, in line with the median diameter at 15 μm below the
requirements, due to the size of electron microscopy in particle control methods have a certain degree of difficulty, it is only as a reference, not included in quality standards. The results shown in Figure 4.
Figure 4 37 ultrafine electron microscopy results (omitted)
In this study, Notoginsenoside R1, ginsenoside Rb1 and
ginsenoside Rg1 for quantitative indicators, in the "Chinese Pharmacopoeia" basis has been improved, a peak time in 35 min can better separation of the sample under test component, this method is more convenient and feasible.
Inspection DiamonsilTMC18 column (150 mm × 4.6 mm, 5μ
m), and Hypersil ODS C18 (150 mm × 4. 6 mm, 5 μm) column on
the samples of different effects of saponins isolated results proved that two different types of color has better spectral separation column.
 State Pharmacopoeia Commission. Chinese
Pharmacopoeia, ? Department of [S]. Beijing: Chemical
Industry Press, 2005:10. Reposted elsewhere in the paper for free download http://