Mechanism underlying the effect of Liuwei Dihuang Pill on osteolysis and osteogenesis induced by titanium particles / 中国组织工程研究

BACKGROUND:At present,a large number of studies have found that Liuwei Dihuang Pill can be used to treat osteoporosis,but there are few related studies on the differentiation and mineralization of osteoblasts induced by wear particles using Liuwei Dihuang Pill. OBJECTIVE:To investigate the positive effect of different concentrations of Liuwei Dihuang Pill-containing serum on titanium particle-induced mouse MC3T3-E1 osteoblast in vitro osteolysis model. METHODS:Drug-containing serum was extracted after oral administration of Liuwei Dihuang Pill.The best concentration of Liuwei Dihuang Pill-containing serum and titanium particles on the viability of MC3T3-E1 cells was screened.MC3T3-E1 cells were divided into three groups.The blank group was given osteoblastic differentiation culture.The model group was given titanium particles(5 μg/mL)ossification culture.The drug-containing serum group was given titanium particles(5 μg/mL)+ Liuwei Dihuang Pill-containing serum(10%,15%and 20%doses).Osteoblast viability was detected by CCK-8 assay.Cell alkaline phosphatase activity was detected by alkaline phosphatase staining.Cell mineralization was detected by silver nitrate(Von Kossa)and alizarin red staining.Expression levels of bone differentiation-related genes Runx-2,Osterix,Ocn,Axin,Alp,and Opn were detected by qRT-PCR.Wnt/β-catenin signaling pathways β-catenin,p-GSK-3β,GSK-3β,Runx2 and Osterix protein expression levels were detected by western blot assay. RESULTS AND CONCLUSION:(1)Liuwei Dihuang Pill-containing serum culture reversed the decrease in alkaline phosphatase activity of MC3T3E-1 cells induced by titanium particles,increased the alizarin red staining and calcification of MC3T3E-1 cells,increased the expression of osteogenesis-related genes in MC3T3E-1 cells,and increased the expression of proteins related to the Wnt/β-catenin signaling pathway.(2)These findings indicate that Liuwei Dihuang Pill-containing serum can reverse the inhibitory effect of titanium particles on the differentiation and mineralization of osteoblasts,upregulate the expression of osteogenesis-related genes,and its mechanism is related to the regulation of Wnt/β-catenin signaling pathway,suggesting that Liuwei Dihuang Pill is expected to become an effective drug for preventing aseptic loosening of artificial joints.

The molecular mechanism of Liuwei Dihuang Pill in the treatment of premature ovarian insufficiency in a mouse model / 中国组织工程研究

BACKGROUND:Most of the formulas for the clinical treatment of premature ovarian insufficiency have evolved from the basic formula of Liuwei Dihuang Pills,and have achieved good therapeutic efficacy.Currently,most of the experimental studies on Liuwei Dihuang Pills focus on morphological observations and physiological and biochemical detection of in vivo animal models,while fewer studies on molecular mechanisms have been reported. OBJECTIVE:To explore the molecular mechanism of Liuwei Dihuang Pills in the treatment of premature ovarian insufficiency based on the receptor gamma coactivator-1 alpha/mitochondrial transcription factor A/reactive oxygen species pathway. METHODS:Premature ovarian insufficiency model was established in mice by intraperitoneal injection of cyclophosphamide 120 mg/kg combined with busulfan 12 mg/kg,and then Liuwei Dihuang Pill suspension was used to intervene in premature ovarian insufficiency mice.After 12 weeks of intervention,the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,anti-Mullerian hormone,8-hydroxydeoxyguanosine,total antioxidant capacity and reactive oxygen species in serum of mice were detected by ELISA method.The morphological changes in mouse ovaries were observed by hematoxylin-eosin staining.The ultrastructure of mouse follicular granulosa cells and the apoptosis of granulosa cell mitochondria were observed by transmission electron microscopy.The expression levels of receptor gamma coactivator-1 alpha and mitochondrial transcription factor A in mouse ovarian granulosa cells were detected by immunohistochemistry. RESULTS AND CONCLUSION:Compared with the model group,serum levels of follicle-stimulating hormone,luteinizing hormone,reactive oxygen species,and 8-hydroxydeoxyguanosine were decreased in the experimental group(P＜0.05),and the levels of estradiol,anti-Mullerian hormone,and total antioxidant capacity were increased(P＜0.05).Hematoxylin-eosin staining showed that in the model group,there were more atretic follicles and corpus luteum forms,some secondary follicles,and interstitial fibrosis and hyperplasia;in the experimental group,a large number of atretic follicles,few corpus luteum forms,primordial follicles were observed at the edges but there were few secondary follicles and no mature follicles.Transmission electron microscopy showed that the organelles in ovarian granulosa cells of mice in the experimental groups were relatively intact.Immunohistochemical results showed that compared with the model group,the expression level of receptor gamma coactivator-1 alpha in the ovarian tissue of mice increased slightly in the experimental group at the 4th week,and there was no significant change at the 8th and 12th weeks.The expression level of mitochondrial transcription factor A in the ovarian tissues of mice in the experimental group was transiently increased at the 4th week,and then slightly decreased,which were all significantly different from those of the model group.To conclude,Liuwei Dihuang Pills inhibit ovarian granulosa cell apoptosis in mice with premature ovarian insufficiency to a certain extent through the receptor gamma coactivator-1 alpha/mitochondrial transcription factor A/reactive oxygen species signaling pathway,thereby improving the endocrine function of the ovary,enhancing the antioxidant capacity,and attenuating the degree of oxidative stress damage.

Liuwei Dihuang Pill () Treats Postmenopausal Osteoporosis with Shen (Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the mechanism of Liuwei Dihuang Pill (, LDP) in treating postmenopausal osteoporosis (PMOP) with Shen (Kidney) yin deficiency.</p><p><b>METHODS</b>In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group (Group A), PMOP Shen-yang deficiency group (Group B), PMOP without Shen deficiency group (Group C), and control group (Group N). Real-time polymerase chain reaction (RT-PCR) and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 (CLCF1), ankyrin repeat and SOCS box containing 1 (ASB1), and prokineticin 2 (PROK2) genes and the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway.</p><p><b>RESULTS</b>The mRNA (P<0.05) and protein (P<0.01) expression levels of the CLCF1 gene in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gene were obviously up-regulated (P<0.01). After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated (both P<0.01), and the average bone density of the top femur had significantly increased (P<0.05). In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3.</p><p><b>CONCLUSIONS</b>The CLCF1 gene is an important gene associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gene expression and activation of the JAK/STAT signaling pathway.</p>

Contribution of Borneolum syntheticum to the Intervention Effect of Liuwei Dihuang Pill () on Experimental Retinal Degeneration / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the contribution of Borneolum syntheticum to the intervention effect of Liuwei Dihuang Pill (, LDP) on experimental retinal degeneration, and initially investigate the mechanism of Borneolum syntheticum as meridian-lead-in drug.</p><p><b>METHODS</b>A total of 180 sodium iodateinduced retinital degeneration rats were randomly divided into three groups, including distilled water group, LDP group, and LDP+Borneolum syntheticum (LDP+BS) group. Twenty normal rats were fed regularly without any treatment as normal control. On day 7 and 14 after treatment, histopathological study and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) test were performed to evaluate the retinopathy. Claudin-5 expression at blood-retina barrier (BRB) was detected by Western blot at different time points from 0.5 to 8 h after gavage.</p><p><b>RESULTS</b>On day 7 and 14 after treatment, the retinal lesion grades were significantly different among the three groups (P<0.05). The grade in the LDP+BS group was significantly less than the LDP and distilled water groups (both P<0.05), no significant difference was observed between the LDP and distilled water groups (P>0.05). The apoptosis rates in the LDP+BS group was significantly less than the distilled water and LDP groups (both P<0.05), while there was no significant difference between LDP and distilled water groups (P>0.05). Expression of claudin-5 in LDP+BS group was significantly less than the other two groups at 0.5, 1 and 2 h after gavage (P<0.05). There was no apparent difference among the three groups at 4 and 8 h after gavage (P>0.05).</p><p><b>CONCLUSION</b>Borneolum syntheticum could strengthen the effect of LDP on experimental retinal degeneration, indicated that Borneolum syntheticum might play the role of meridian-lead-in drug in the formula. The mechanism may be due to Borneolum syntheticum could promote the physiologically openness of bloodretina barrier through transiently affecting the expression of claudin-5.</p>

The Clinical Experience of Professor HE Ruoping in the Treatment of Perimenopausal Syndrome / 浙江中医药大学学报

[Objective] To summarize the academic views and clinical experience of Professor HE Ruoping in the treatment of peri menopausal syndrome by using traditional Chinese medicine. [Method] From the clinical treatment, collecting the clinical information of the teacher, research and discuss the traditional Chinese medicine academic ideas and clinical experience of teacher HE in the treatment of perimenopausal syndrome, to summarize the traditional Chinese medicine of perimenopausal syndrome in dialectical characteristic.And further explain the characteristics of the treatment of teacher HE through case analysis. [Result] Teacher HE thinks that the main pathogenesis of perimenopausal syndrome is the deficiency of kidney Yin, and divides the patients into stagnation of liver-qi type, Qi stagnation and blood stasis, disharmony between heart and kidney, heart and spleen deficiency type, yin-yang deficiency type, a total of five kinds of syndrome types, according to their clinical manifestation.When confronted with such patients, teacher HE often chooses Liuwei Dihuang Pills to tonify kidney-yin and relieve liver for smooth Qi, promoting qi to activate blood, restoring normal coordination between heart and kidney, invigorating heart and spleen, tonify both Yin and Yang, gives patients more grooming solution at the same time to adjust their physical and mental state as a whole, so the patients could receive remarkable effect and avoid the potential risks of hormone replacement therapy.The case in this article has obtained good treatment results. [Conclusion] Teacher HE treats perimenopause syndrome by tonifying kidney-yin and taking accompanied symptoms and signs into account meanwhile, which has significant clinical effect and the value of popularization and application.

Investigation of herb-drug interaction between Liuwei Dihuang Pills and nifedipine based on rat liver microsomes and HPLC fingerprints / 中国药科大学学报

@#A new method based on rat liver microsomes and chromatographic fingerprint comparison was established to investigate the possible herb-drug interactions between Liuwei Dihuang Pills(LDP)and nifedipine(NIF). LDP and NIF were incubated simultaneously with rat liver microsomes at 37 °C for 60 min in a shaking water bath. The separation was achieved on a C18 column using 0. 1% formic acid solution and acetonitrile as mobile phase with a liner gradient program. The flow rate was set at 1. 0 mL/min. Detection was achieved by UV light at 240 nm. To evaluate the interactions between LDP and NIF, the similarity of the fingerprinting chromatograms before and after co-incubation was calculated by similarity evaluation system for chromatographic fingerprint of TCM. Furthermore, pharmacokinetic experiments in rat suggested that there was no significant difference in the pharmacokinetic parameters of the typical components in LDP.

Clinical Observation of Liuwei Dihuang Pill Combined with TCM Prescription Acupoint Application in the Treatment of Type 2 Diabetic Constipation / 中国药房

OBJECTIVE:To obverse the efficacy and safety of Liuwei dihuang pill combined with TCM prescription acupoint application in the treatment of type 2 diabetic constipation. METHODS:120 patients with type 2 diabetic constipation were random-ly divided into control group(60 cases)and observation group(60 cases). All patients received controlling blood pressure and regu-lating blood lipids drugs,diet control combined with exercise,and fixed dose of Metformin hydrochloride tablet(0.5 g/times,2 times a day)and Acarbose tablet(100 mg/times,3 times a day);based on it,control group received 5 mg Mosapride citrate tablet, 3 times a day;observation group additionally received 8 Liuwei dihuang pills(3 times a day)combined with TCM prescription acu-point application (in double Tianshu Point,double Yongquan point,double Shenque Point and double Zusanli). 7 d was regarded as a treatment course,and it lasted 3 courses. Clinical efficacy,blood glucose [2 h postprandial glucose(2 h PG),blood glucose (FPG)],the clinical symptom indexes(defecation interval time,each defecation time,weekly defecation frequency)before and af-ter treatment,and the incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than in control group,and the difference was statistically significant (P0.05). After treatment,2 h PG and FPG in 2 groups were significantly lower than before,and obser-vation group was lower than control group;defecation interval time and each defecation time were significantly shorter than before, and observation group was shorter than control group;weekly defecation frequency was significantly higher than before,and obser-vation group was higher than control group,The differences were statistically significant (P0.05). CONCLUSIONS:Liuwei dihuang pill combined with TCM priscription acupoint ap-plication shows good efficacy in the treatment of type 2 diabetic constipation. It can effectively improve constipation,reduce defeca-tion interval time and each defecation time,increase weekly defecation frequency and have no influence on blood glucose control with small adverse reactions and high safety.

Correlation of powder particle size, cell wall-breaking, rate and in vitro dissolution rate of Liuwei Dihuang Pill / 中草药

Objective: To investigate the relationship among powder particle size, cell wall-breaking rate, and dissolution of Liuwei Dihuang Pill (LDP) in vitro. Methods: Particle size and cell wall-breaking rate of four kinds of LDP were measured by laser particle size instrument and optical microscope. The loganin and paeonol were taken as indexes, the feature of dissolution for the above four kinds of LDP was examined by grouting method. Results: There were significant differences in the distribution of particle size, wall-breaking rate, and dissolution among different crushing degrees of LDP. With the decrease of the powder particle size, the wall-breaking rate and relative cumulative dissolution of loganin and paeonol increased. The rate of characteristic cell wall-breaking in the micro powder of Corni Fructus, Alismatis Rhizoma, and Moutan Cortex could reach 100%. Conclusion: Moderately changing degree of smash could cause cell wall-breaking rate changes, so as to control effective ingredient the dissolution rate and extent of Chinese medicine.

Clinical study on addition treatment of liver -kidney yin deficiency and postmenopausal osteoporosis with Liuwei Dihuang pill / 中国基层医药

Objective To study the clinical effects of addition treatment of liver-kidney yin deficiency and postmenopausal osteoporosis with Liuwei Dihuang pill.Methods 120 patients suffering from postmenopausal osteopo-rosis who were determined to be subject to liver -kidney Yin deficiency by doctors of traditional Chinese medicine when they received treatment at the outpatient clinic of traditional Chinese medicine department of our hospital in the period from February 2010 to june 2013 were divided into three groups:group A,B and C,with the random number table.For group A:provided the honeyed pills made up by supplemented six -ingredient rehmannia pill,and taken two pills per day continuously for six months;For group B:provided 5mg of zoledronic acid injection for intravenous drip infusion;For group C:provided traditional Chinese medicine honeyed pills and zoledronic acid for treatment at the same time;all of the three groups take Calcium carbonate D3 at the same time;The three groups of cases until the 12th month were observed and followed -up.The clinical efficacy of three groups were compared.Results A,C group 3 months after treatment with estradiol levels compared with before treatment,the difference was statistically significant (t =2.54,P =0.05,t =2.07,P =0.06).A group 3 months after treatment,the VAS scores at 6 months respectively (3.75 ±1.32),(2.06 ±1.53)points before treatment,and the differences were statistically significant(t =10.18, P =0.00,t =9.26,P =0.00).A group 3 months after treatment,the SF -36 scores at 6 months were (90.82 ± 12.38),(105.82 ±9.06)% respectively,compared with that before treatment,the differences were statistically significant(t =8.40,P =0.00,t =5.21,P =0.00).A group 6 months after treatment of lumbar 1 -4 bone density-were (0.85 ±0.07)g/m2 ,and before treatment[(0.76 ±0.07)g/m2 ]comparison,the difference was statistically significant(t =7.56,P =0.00);A group 6 months after treatment of Wards bone density compared with before treatment,the difference was statistically significant(t =8.07,P =0.00).C group 6 months after treatment of lumbar 1 -4 bone density(0.91 ±0.07)g/m2 ,and B Group[(0.88 ±0.05)g/m2 ]comparison,the difference was statistical-ly significant(t =27.19,P =0.00);C group after treatment,the bone mineral density of Wards area for 6 months were (0.62 ±0.04)g/m2 .The difference was statistically significant (t =26.22,P =0.00),compared with the B group. The TCMsyndromes of TCMin the 6 months after treatment in A group were (5.42 ±2.13),and the difference was statistically significant (t =13.29,P =0.00),compared with the B Group (10.75 ±4.08).(t =13.29,P =0.00). Compared with B group,the difference was statistically significant(t =26.22,P =0.00).A group after the treatment of TCMsyndrome score was 6 months(5.42 ±2.13),and B Group[(10.75 ±4.08)],the difference was statistically significant(t =13.29,P =0.00).Conclusion The addition of Liuwei Dihuang pill can improve BMD,alleviate the ostealgia and improve life quality of patients suffering from postmenopausal osteoporosis,and has no obvious adverse effects.It can enhance curative effects through using with diphosphonate.

Improvement of quality control method of liuwei dihuang pills / 中国药学杂志

OBJECTIVE: To improve the quality control method of Liuwei Dihuang Pills. METHODS: Morroniside and loganin from processed Fruetus Corni were identified by TLC. Morrniside, loganin and paeonol in Liuwei Dihuang Pills were determined by HPLC. The determination was performed on a Phenomenex Gemini 110A C18 column (4.6 mm×250 mm, 5 μm) maintained at 40℃ with gradient elution of acetonitrile (A) and 0.3% phosphoric acid aqueous solution (B) at a flow rate of 1 mL·min-1. The detective wavelength was set at 240 nm for morroniside and loganin and 274 nm for paeonol. RESULTS: The TLC spots were clear. Morrniside, loganin and paeonol showed good linear relationship in the range of 0.02078-1.039, 0.02034-1.017, and 0.04576-2.288 μg, respectively. The average recoveries (n=9) were 99.97%-100.99% with RSDs less than 2.3%. CONCLUSION: The method is simple, rapid, accurate and can be used to control the quality of Liuwei Dihuang Pill more effectively.

Establishment of quantitative determination methods of total glycoside groups composition in Chinese materia medica by differential anthrone-sugar hydrazone / 中草药

Objective: To establish a method for the determination of total glycosides in Chinese materia medica (CMM)-differential anthrone-sugar hydrazone colorimetry. Methods: The content of total glycosides (including sugar) in CMM was determined by anthrone colorimetry, and then sugar hydrazone-sulfuric acid colorimetric method was used to determine the content of reducing sugars. The content of total glycosides came from the two subtraction. Results: Anthrone colorimetric reaction was completed within 6 h, detection wavelength was 620 nm, linear range was 80-180 μg/mL, correlation coefficient R2 = 0.994, and recovery was 99.2%, RSD was 1.35%. Sugar hydrazone-sulfuric acid color reaction method was completed within 4 h, detection wavelength was 412 nm, linear range was 2-12 mg/mL, correlation coefficient R2 = 0.991, and recovery was 99.5%, RSD was 1.98%. The total glycosides contents in five Chinese patent medicines such as Buyang Huanwu Decoction, Liuwei Dihuang Pill, Andrographis Tablet, Astragalus Injection, and Pachymaran Oral Liquid were 15.1, 26.0, 6.68 mg/g, and 21.3, 16.9 mg/mL. Conclusion: This method is firstly established for the quantitative determination of total glycosides in CMM, and the ingredient group analysis method is established by anthrone colorimetry and sugar hydrazone-sulfuric acid colorimetric method. The methods have better accuracy, and can be used for the determination of total glycosides ingredients group of CMM.

Exploring Clinical Indications of Liuwei-Dihuang Pill through Text Mining / 世界科学技术-中医药现代化

Liuwei-Dihuang Pill is a famous medicine in the clinical practice of traditional Chinese medicine (TCM), and it is important and interesting to explore its new clinical indications. This study is based on the data downloaded from the SinoMed by querying on term of Liuwei-Dihuang Pill. By applying the data slicing algo-rithm, we achieved outstanding symptoms associated with Liuwei-Dihuang Pill. Examining these symptoms, besides those which are widely known as common knowledge, some other potential symptoms are found. These potential symptoms are aphtha, (sexual) dysfunction, constipation, and heel pain which might be taken as new clinical indi-cations of Liuwei-Dihuang Pill.

Quality Standard of Liuwei Dihuang Pill(Concentrated Pill) / 中国药房

OBJECTIVE:To establish the quality standard of Liuwei dihuang pill (concentrated pill). METHODS:Radix Rehmanniae Preparata,Dioscorea opposita,Poria cocos and Alisma orientalis in Liuwei dihuang pill (concentrated pill) were identified by TLC and the contents of paeonolum and loganin were determined by HPLC.RESULTS:The TLC spots were clear and well-separated yet free of interference of negative sample.The linear ranges of paeonolum and laganin were 0.100 6～1.006 ?g (r=0.999 9) and 0.309 6～2.322 ?g(r=0.999 6),respectively,and their recovery rates were 98.2%(RSD=2.3%,n=6) and 95.3%(RSD=0.42%,n=6),respectively.CONCLUSION:The established standard is applicable for the quality control of Liuwei dihuang pill.

Clinical Application and Study on Liuwei Dihuang Pill Composition / 浙江中医药大学学报

It explores the disease range of Liuwei Dihuang Pill from formulae source,ancient clinical application,modern study and application angles,combining with modern diseases,to define its multi-system targets,main pharmaceutical function and effective components by seeking for common mechanism.

Discussion on Quantitative Standard of the Pill and Granule of Liuwei Dihuang in 2005 Edition of China Pharmacopoeia / 中国药房

OBJECTIVE:To discuss the problems of quantitative standard and method for the pill and granule of Liuwei Dihuang in 2005 edition of China Pharmacopoeia.METHODS:The quantitative indicators,determination methods,and content standards established in 2005 edition of China Pharmacopoeia were compared between the pill and granule of Liuwei Dihuang,these indices were also compared with those in 2000 edition of China Pharmacopoeia.RESULTS &CONCLUSIONS:The quantitative indicators,determination methods,and content standards set for pill and granule of Liuwei Dihuang are not consistent between the horizontal and vertical comparisons,it is suggested that these indices should be revised when China Pharmacopoeia is reprinted.

Attenuation Action of Liuwei Dihuang Pill on Aristolochia Fangchi-induced Toxicity / 中国药房

OBJECTIVE: To study the attenuation action of Liuwei dihuang pill on Aristolochia Fangchi-induced toxicity through determination of the content of Aristolochia acid A in Aristolochia Fangchi. METHODS: After mixed decoction of Aristolochia Fangchi with Liuwei dihuang pill,the content of Aristolochhia acid A was determined by HPLC and the content change was observed. RESULTS: Aristolochia acid A content in Aristolochia Fangchi was markedly decreased after intervention by Liuwei dihuang pill. CONCLUSION: The attenuation action of Liuwei dihuang pill is related to the downregulation of Aristolochhia aicd A content in Aristolochia Fangchi.

Content Determination of Paeonol in Liuwei Dihuang Pill by SPE - HPLC / 中药新药与临床药理

Objective To develop SPE - HPLC method for the assay of paeonol in Liuwei Dihuang Pill (LDP). Methods Separation was performed on a Hypersil ODS(2) column (250 mm?4. 6 mm, 5?m) . The mobile phase consist of methanol - 2 % acetic acid (55 :45, v/v) . The flow rate was 1 mL/min. The UV detection was set at 275 nm. Results Paeonol had a good linear relation in the range of 0. 03998 ~ 0. 7996 ug, the average recovery of honeyed bolus of LDP was 102. 9 % and the RSD was 1.2%; the average recovery of concentrated bolus of LDP was 101. 5 % and the RSD was 1.8%. Conclusion The method is simple, sensitive and rapid and it can be used to determine the paeonol content of LDP.

Quality evaluation of Liuwei Dihuang Pill / 中成药

AIM: To compare the quality of Liuwei Dihuang Pill among the pharmaceutical companies by determining the content and the dissolution of paeonol of Liuwei Dihuang Pill. METHODS: A method of determining paeonol and the dissolution from Liuwei Dihuang Pill was establishod. HPLC was used with methanol-water (70 ∶30) as a mobile phase and detection wavelength was at 274 nm. The flow rate was 1.0 mLmin, injection volume was 20 ?L. The percentage of dissolution was determined in the 30% methanol- 0.1 mol1 HCL solution. RESULTS: A good linearity was over the weight range of 1.4 ?g-140 ?g, the average recovery was 97.38% , the RSD for reproducibility was 1.44% (n=6). The parameter of Tianjin’s production was T_ 50 = 156.77 , T_d= 256.55 ,M= 0.734 and the parameter of Beijing’s production was T_ 50 = 110.66 , T_d= 197.19 , M= 0.583 . CONCLUSION: The method is simple, and can be used to determine the content and the percentage of dissolution of paeonol in Liuwei Dihuang Pill. The experiment indicates that the percentage of dissolution has no obvious distinction in products from different manufactures; but has obvious distinction in the batches of same manufacture.

Determination of ursolic acid and oleanolic acid in Liuwei Dihuang Pill from different manufacturers by HPLC-PAD / 中成药

AIM:To observe the contents of ursolic acid and oleanolic acid in Liuwei Dihuang Pill(Radix Rehmanniae praeparata,Rhizoma Dioscoreae,Fructus Corni,Rhizoma Alismatis,etc.) from different manufacturers and explore their effects on quality of Liuwei Dihuang Pill.METHODS:Ursolic acid and oleanolic acid in Liuwei Dihuang Pill were determined by HPLC-PAD method using Kromasil C_ 18 column(4.6 mm?250 mm,5 ?m) as chromatographic column,methanol-water-phosphate(88∶12∶0.02) as the mobile phase at the rate of 1.0 mL/min.RESULTS:6.462-0.718 ?g of ursolic acid and 3.132-0.348 ?g of oleanolic acid presented a good linear relationship.The average recoveres were 99.73% and 97.59%,RSD were 1.5% and 1.7%,respectively.There were significant differences in the contents of ursolic acid and oleanolic acid in Liuwei Dihuang Pill from four different manufacturers.CONCLUSION:The method is simple,rapid and feasible to set up determination of ursolic acid and oleanolic acid in Liuwei Dihuang Pill by HPLC-PAD.Although the contents of ursolic acid and oleanolic acid have significant differences in different manufactures,the results show good consistency in the content variation of the two acids.

Quality differential analysis of Liuwei Dihuang Pill / 中成药

AIM:To compare the quality of Liuwei Dihuang Pill produced by different pharmaceutical companies by determining the content and the in vitro dissolution of paeonol and loganin of Liuwei Dihuang Pill.METHODS:The HPLC method was used for the assay of paeonol and loganin in Liuwei Dihuang Pill.The standard dissolution test was used to investigate the in vitro dissolution of paeonol and loganin.RESULTS:The content of paeonol and loganin from different manufacturers differed significantly.Six products differed significantly from one another in their in vitro dissolution(P