Study On Pharmaceutical Research And Acute Toxicity Test Of Tangshenkang Pill

Tang shen kang Pill consists of eleven kinds of traditional Chinese medicine,including Astragalus,Cornus Officinalis,Concha Ostreae,SchisandraChinensis,CortexEucommiae,SemenEeuryales,Cortex Phellodendri,Fructus Rosae Laevigatae,Leonurus japonicus,O?theca Mantidis and Radix Glycyrrhizae.It has the effect of tonifying qi and yin,tonifying kidney,arresting seminal emission.It has been clinically used in the treatment of early and meta-phase diabetic nephropathy.Moreover,for the convenience of the patients'usage,Tang shen kang Pill,a hospital salted recipe,was made into concentrated honeyed pill,which is widely used to clinic and the curative effect is definite.However,its preparation process is simple while the methods of quality control are not objective.In order to effectively control the quality of this preparation,researches on preparation technologies,quality standards,stability investigations have been carried out.At the same time,in order to evaluate the clinical medication safety of the Tang shen kang pill,acute toxicity test on the mice was performed.Part 1 Study on the technology of preparation processAccording to the physical and chemical properties of various kinds of traditional Chinese medicine in Tang shen kang pill and clinical practice,3kinds of Chinese medicine like Leonurus japonicus,Fructus Rosae Laevigatae and O?theca Mantidiswere extracted by water and 8 kinds of Chinese medicine were crushed into fine powder,processed into condensed honeyed pills including Astragalus,Concha Ostreae,Cortex Eucommiae,Cornus Officinalis,Fructus Schisandrae Chinensis,Semen Eeuryales,Cortex Phellodendri and Radix Glycyrrhizae.As index of the extract rate,the water addition,the times for decoction and decocting duration were optimized by L9?3?⁴ orthogonal test to comprehensively evaluate the water extraction process of Leonurus japonicus,Fructus Rosae Laevigatae andO?theca Mantidis.The optimal extraction process was that herbs were decocted for two times in the water,the amount of water was equal to ten times the dose of medicine,and 1.5 hours each time.As indexes of the pulverization complexity and the yield of powder,the pulverization technology was studied by comparative test to comprehensively evaluate the pulverization process of Astragalus,Concha Ostreae,Cortex Eucommiae,Cornus Officinalis,Fructus Schisandrae Chinensis,Semen Eeuryales,Cortex Phellodendri and Radix Glycyrrhizae.The optimal pulverization technology was grinded by materials mixed and then passed through the 100 mesh.As indexes of product yield and colors,the calendaring techniques of honey were studied by comparative test also.The results showed that while the water addition was a quarter of the amount of honey is the optimal technology.As indexes of disintegration time and water content of honeyed pills,the extracts proportion,the dosage of honey and the mixing temperature were optimized by L9?3?⁴ orthogonal test to comprehensively evaluate the forming technology of concentrated honeyed pills.Results showed that the water content and the disintegration time of the condensed water honey pill could meet the requirement when the relative density is between 1.13%and 1.14%,refined honey amount is 0.4 times the weight of the powder and the mixing temperature is 70 degrees Celsius.Finally,the technologies of water decotion and concentrated honeyed pills'formation were simultaneously carried out for validation by lab test,and the results showed that the optimal technologies are stable and feasible.Part 2 Study on quality standardsTLC is adopted to identify Astragalus,Cornus Officinalis and Schisandra Chinensis in Tang shen kang pill.The test results showed that the chromatogram of sample correspond the same color spots with that of reference crude herb or reference substance at corresponding location,without the interference of negative control.HPLC is adopted to measure the content of berberine hydrochloride in Tang shen kang Pill with the chromatography conditions as followings:Phenomenex Gemini-NX C₁₈8 chromatography column?250 mm×4.6 mm,5?m?;mobile phase of acetonitrile-0.1%phosphate?25:75?,flow rate of 1.0 mL·min^-1;the detection wavelength of270 nm;the column temperature of 30?.The number of theoretical plates should not be less than 5000 when calculated by berberine hydrochloride peak.At the same time,research on methodology has been carried out,which the results showed that the method is reasonable and feasible with good reproducibility and can be used for the quality control of this preparation.When calculated by berberine hydrochloride,the preparation shall contain no less than 1.7%yellow cypress.Part 3 Stability studiesTang shen kang pills were packaged in polyethylene plastic bottles,natural storaged for 12 months at room temperature and storaged for 6 months under the condition of accelerated test.All the detection indicators are in line with the provisions of the product's quality standards,preliminarily suggesting that the product's quality is stable.Part 4 Study on acute toxicity testMice were fed with Tang shen kang pill of maximum concentration and the maximum volume which the mice can withstand in stomach,which means the maximum dose at one time in one day,so as to determine the maximum tolerated dose.By doing acute toxicity test,we observe the safety of Tang shen kang pill.Results:the maximal tolerance dose of Tang shen kang pill which mice can bear by gastric lavage is 211.20 g crude drug/kg,equal to 468times of the adult's clinical daily dosage of 22.50 g.Conclusion:Tang shen kang pill is safe.Therefore it is safe in clinical medication.