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8Study on the quality standards of guiling schisandra chinensis glycyrrhizae
radix decoction agent
Zhang Na
Heilongjiang University Of Chinese Medicine(Heilongjiang province, Harbin city,China)
Abstract:Using TLC method to identify glycyrrhizae radix and schisandra chinensis .HPLC was adopted to evaluate the quality of schizandrin A.The results showed that TLC method could obviously identified glycyrrhizae radix and schisandra chinensis.Schizandrol A had a linear relationship at a range of 0.021~0.189 mg/mL.r=0.9998.The average recovery rate was 98.14%,The relative standard deviation=2.01%.It explains the method developed is stable,reliable and repeatable for guiling schisandra chinensis glycyrrhizae radix decoction agent and the method is reasonable and feasible,and quality control.
Keywords:guilingschisandra chinensis glycyrrhizae radix decoction agent; TLC; HPLC; quality standards
The soup of guiling schisandra chinensis glycyrrhizae radix from the << Synopsis of Golden Chamber >>[1],the prescription was composed of cassia twig,tuckahoe,schisandra chinensis and glycyrrhizae radix.It had effect on moistening lung for resolving fluid retention and washing down QI.We established TLC of schisandra chinensis and glycyrrhizae radix.As well as using HPLC to measure the content of schizandrin A.To control the decoction's quality. 1.Materials and methods 1.1Instruments and reagent HPLC(Waters 1515,American waters co.) Ultrasonic cleaners(MP5200H,ShanXi PengZhan techonoly co.) Electronic Analytical Balance(AB204-N,Mettler Toledo instrumentco.LTD,Shanghai) schizandrin AStandard control substance(National Institute for the Control of Pharmaceutical and Biological Products,NO:110736-200929) 2Results and discussion 2.1schisandra chinensis TLC identification The Reference drug solution:[2]
Weighingthe medicine of Schisandra approximately 1g,with the preparation of reference drug solution by the method sample solution prepared.
Preparation of standard solution:
Taking schizandrol reference substance,adding methanol to make concentration was 1mg/ml solution as the reference solution.
Preparation of negative test sample solution
Preparation process made without Schiandra negative sample,the test sample solution was prepared by the method for producing the negative test sample solution.
TLCReferring to TLC[3] (Chinese Pharmacopoeia 2010 edition an Appendix VI B)test,to draw the above test and negative test sample solution of8~10^l, The Reference drug solution and Preparation of standard solution of2~4pl.Respectively pointed in the same GF254 TLC plate,with toluene - ethyl acetate (9:4) as the agent,started out,drying,UV light(254nm),under review. 2.2 glycyrrhizae radix TLC identification Preparation of test sample solution:
Taking this product 5ml,set separating funnel,add N-butanol saturated with water 20ml, SeperatingN-butanol alcohol extracts,and using the water of N-butanol saturated extraction two times,10ml everytime, the part ofN-butanol evaporated to dryness,the residue was dissolved in methanol 1ml,as the test solution.
The Reference drug solution:Weighingthe medicine of glycyrrhizae radix approximately 1g,with the preparation of the test sample solution prepared by the method reference drug solution.
Preparation of negative test sample solution
Preparation's process made without glycyrrhizae radix negative sample,the test sample solution was prepared by the method for producing the negative test sample solution.
TLC According to TLC(Chinese Pharmacopoeia 2010 edition an Appendix VI B)test,to draw the above test and negative test sample solution of15pl, The Reference drug solution of5~8^l.Respectively pointed in the same G TLC plate,with ethyl acetate - methanoic acid - glacial acetic acid -water (15:1:2:2) as the agent,started out,drying,UV light(365nm),under review.
Conclusions: the above photos indicated the test solutions had the same fluorescent spots,in comprison with the corresponding position of the reference drug chromatographys.Nevertheless the negative control solution hadn't the spots.
3Content determination
3.1 Schraandrol A' content determination and Results
Preparation of the schisandra chinensis sample solution
Weighing the powder about 0.1261g in the 10ml volumetric flask and adding methanol 8ml.Ultrasonic processing(power 250W, frequency 20kHz)20minutes.Taking out and adding methanol to the lable.shaking and filtration.Finally the filtrate obtained.
Preparation of the standard solution
Weighing precision the Schizandrol A standard sample 5.13mg in the 25ml volumetric flask and adding methanol to the lable.Shaking untill to reslove completely.Finally obtained.(C=0.21mg)
HPLC method. Drawing precision two solutions above 10^l respectively and injecting into HPLC.Calculating the content of Schizandrol A in schisandra chinensis.The results were shown in Table 3-1.Analysis: herbs' measurement results comply with the Chinese Pharmacopoeia 2010 version of schisandra chinensis content requirements.
3.2Establishing the method of Schraandrol A' content determination
Chromatographic conditions Column:
Agilent Extend ODS Reversed phase column(4.6mm*250mm, 5p,m);the mobile phase:methol-water(11:9);Determine wavelength:254nm;Flowing rate: 1.0ml/min;Column temperature:room temperature;The injection volume was 10pl;n>=2000.
3.2.1Preparation of standard curve
Respectively measuing 3.1 Standard solution of Schizandrol A(C=0.21mg/ml) 0.5ml, 1.5ml, 2.5ml, 3.5ml, 4.5ml in5ml volumetric flask.Adding methanol to the lable.Respectively drawing 10^1 and injecting into HPLC.Finally measured the peaks area.As well as set the sample volume(mg)as the abscissa and measured peaks area as the vertical axis.Obtain the regression equation Y=6.77x104X-1.68x104,r=0.9998 NO3:Standard curve
S mg/ml
Conclusion: Schizandrol A'peak area reveal a goog linear relationship between0.21x10
1~1.89x10-1 mgml-1
NO4. Linear relationship research
standard (mg/ml) peak area r regression equation
0.21x10-1 49544 0.63x10-1 185331
1.05x10-1 370451 0.9998 Y=6.77X104X-1.68X104 1.47x10-1 555788 1.89X10-1 741213
3.2.2Precision test
Drawing precision sample (NO:20100601) [4-6]3ml.Producing the sample solution in according with the method of2.2 sample solution.Drawing the solution 10^l and injecting into HPLC continuous six times.HPLC spectra was photo6.calculated the content and RSD.The sequence was the form5. NO5.Precision investigation
NO sample volume peak area Mean RSD
(ml) (A) (A) (%)
1 3 321045
2 3 311916
3 3 319475 316779 1.26
4 3 319006
5 3 311745
6 3 317487
3.2.1Preparation of standard curve
Respectively measuing 3.1 Standard solution of Schizandrol A(C=0.21mg/ml) 0.5ml, 1.5ml, 2.5ml, 3.5ml, 4.5ml in5ml volumetric flask.Adding methanol to the lable.Respectively drawing 10^1 and injecting into HPLC.Finally measured the peaks area.As well as set the sample volume(mg)as the abscissa and measured peaks area as the vertical axis.Obtain the regression equation Y=6.77x104X-1.68x104,r=0.9998 NO3:Standard curve
S mg/ml
Conclusion: Schizandrol A'peak area reveal a goog linear relationship between0.21x10
1~1.89x10-1 mgml-1
NO4. Linear relationship research
standard (mg/ml) peak area r regression equation
0.21x10-1 49544 0.63x10-1 185331
1.05x10-1 370451 0.9998 Y=6.77X104X-1.68X104 1.47x10-1 555788 1.89X10-1 741213
3.2.2Precision test
Drawing precision sample (NO:20100601) [4-6]3ml.Producing the sample solution in according with the method of2.2 sample solution.Drawing the solution 10^l and injecting into HPLC continuous six times.HPLC spectra was photo6.calculated the content and RSD.The sequence was the form5. NO5.Precision investigation
NO sample volume peak area Mean RSD
(ml) (A) (A) (%)
1 3 321045
2 3 311916
3 3 319475 316779 1.26
4 3 319006
5 3 311745
6 3 317487
3.2.3Accuracy test(recovery test)
Mixturing the samples (NO:20100601)about 1.5ml with C=0.21mg/ml standard solutions 2ml.Repeating six times. Producing the sample solution in according with the method of sample solution. Drawing the solution 10^1 and measuring the peaks area.Finally calculating recoveries.HPLC spectra was photo6.The results were shown in Form6.
Recovery (%) = (measure content—content of sample) /adding standard content* 100% NO.6 accuracy investigation
NO sample volume content add measure recobery average RSD
(ml) (mg) (mg) (mg) (%) (%) (%)
1 1.5 0.48 0.41 0.84 100.54
2 1.5 0.48 0.41 0.83 98.51
3 1.5 0.48 0.41 0.82 96.64
4 1.5 0.48 0.41 0.82 95.56 98.14 2.01
5 1.5 0.48 0.41 0.83 100.17
6 1.5 0.48 0.41 0.83 97.39
Conclusion:The recovery of Schizandrol A are between 95% and 105%.RSD<5%.The method was revealed reasonable and reliable by the figures.
3.2.4Establishing the content of Schizandrol A in the samples Preparation of the standard solution
Weighing precision the Schizandrol A standard sample 5.13mg in the 25ml volumetric flask and adding methanol to the lable.Shaking untill to reslove completely.Finally obtained.(C=0.21mg/ml)
Preparation of test sample solution: Taking this product 3ml and evaporating to dryness. The residue was dissolved with methanol in the 25ml volumetric flask and adding methanol to the lable.Finally the sample solution was obtained. The method of measurement
Drawing precision above two solutions 10^l respectively and injecting into the HPLC . determinating of the peak area, Calculating the content of Schizandrol A according to the following formula.
3.2.5Content of sample determination
According to the methods above.three batches of samples for test and three parallel determinations in every batch of samples.The results were shown in Form 7.
NO.7Content of Schizandrol A in sample
NO. content (mg ml-1) average (mg ml-1) RSD (%)
20100601 0.29
20100602 0.28 0.29 2.01
20100603 0.29
Conclusion:Through the determination of several batches of samples,and basement on the rules of schisandra chinensis (Chinese Pharmacopoeia 2010 edition an Appendix VI B). The content of Schizandrol A>=0.4%.water<=16%.Finally,the decotion was ruled the content of Schizandrol A C24H32O7 no less than 0.20mg/ml. References
[1]Zhang ZJ,zhongjing encyclopedia on golden chamber of themethodology[M],Beijing:TCM Ancient Books Publishing House,2010:139-140.
[2]Fan ZW,Tian M,Liang YX,et al.Schisandra medicinal HPLC fingerprint analysis of different extraction methods[J].Journal of Northeast Normal University:Nature science,2004,36(2):50-54.
[3]Pharmacopoeia editorial board.The people's republic of China pharmacopoeia 2010 edition[M].Beijing:China press of Traditional Chinese Medicine,2010:61.
[4]Jiang ZJ,Lu Y,Chen DF.Kadsura TLC identification and quality standards research[J].Lishizhen Med Mater Med Res,2010,21(11):2899-2901.
[5]Cao JH.Traditional Research of Chinese medicine decoction quality standard[J]. Lishizhen Med Mater Med Res,2006,17(6):1002-1003.
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Different origin astragalus microscopic identification
Zhang yanhe Ma wei* Gong he Zhang leiming College of Pharmaceutical Sciences, HeilongjiangUniversity of Chinese Medicine, Harbin China
Abstract: The purpose of perfecting Astragalus membranaceus(Fisch)Bge and Astragalus membranaceus (he.) Bge. Var. mongholicus(Bge. )Hsiao identification standard. Methods character identification and microscopic identification of the different morphology of Astragalus and organizational structure to carry out a comparative study. The results of the of different Astragalus Medicinal parts, Appearance and cross-section of organizational structure there are obvious differences. Conclusion ecological environment have a larger impact on the growth of Astragalus Radix morphology of the different growth environment and there are significant differences in the organizational structure.
Keyword Astragalus; traits to identify ;microscopic identification
Introduction
Astragalus as bean secco plant Mongolia Astragalus Astragalusmembranaceus (he) Bge. Var. Monghoicus (Bge) Hisao or membrane pod Astragalus membranaceus Bge (he), the dry root. A. memebranaceus is used in traditional Chinese medicine, especially to give a medical treatment for diabetes.In Western herbal medicine,Astragalus is primarily considered to be a tonic forenhancing metabolism and digestion. It is consumed as a tea or soup made from the (usually dried) roots of plants,also often in combination with other medicinal herbs. In addition, Astragalus is traditionally used to strengthen theimmune system and the healing effect of wounds and injuries. [1]. Modern research analysis, Astragalus membranaceus contain a variety of saponins, flavonoids, polysaccharides, as well as amino acid, linoleic acid, alkaloids, choline and complex chemical composition [2-3]. Astragalus in protecting blood vessels, and cardiac muscle, liver, lower serum transaminase, immune regulation, blood rheology, regulate blood sugar diabetes, kidney disease, enhance the role of resist free radical attacks. And there is the potential analgesic, sedative and hemostatic function [4-7].Astragalus memebranaceus agriculture in Shanxi, Inner Mongolia, Hebei, Shaanxi, Gansu and other provinces and regions. As the most abundant wildlife resources the
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Corresponding auther Phone: 0451-82193430
E-mail: [email protected]
