Studies On Determination Method And Quality Criteria Of Residua Of Macroporous Resins

The macroporous resins is used widely in the field of pharmaceutics , such as the refinement and purification of the effective fraction or constituents from Chinese Ttaditional Medicine and quality control. It is a very effective extraction, separation and purfication method. There are some questions in its application, such as the criterion of residua and the pharmaceutic security. The following researches in this thesis are studied the determination method of the residual solvents and the evaluation methods of its pharmaceutic technology. 1 Study on the pretreatment methods of macroporous resins .Selection of pretreatment methodsThe fist method :Alkali-EtOH method:Dip the macroporous resin with 2BV 5% NaOH in 80℃ water bathe for 6 hours, elute the resin with 80℃ distilled water until the elute is neutral, filter the water and dip the resin with 2BV 95%EtOH for 24h, elute the resin with 95% EtOH until the absorptance of the eluent is less than 0. 03 at 254nm, elute with distilled water until the eluent without EtOH.The second method '.Ultrasonic method :Dip the macroporous resin with 2BV EtOH for 24h, filter the EtOH, Dip the resin with 1BV EtOH , ultrasonic extraction until the absorptance of the eluent is less than 0.03 at 254nm, elute with distilled water until the eluent without EtOH.The third method:Evaporation method: The macroporous resin evaporate on 80°C water bathe for 4 hours, Dip resin with 2BV EtOH for 24h, filter the EtOH , Dip the macroporous resin with 2BV 5% NaOH in 80℃ water bathe for 6 hours, such as the fist method.The fourth method: Vacuum evaporation method:The macroporous resin evaporate in 80°C vacuum container for 4h, Dip resin with 2BV EtOH for 24h, filter the EtOH , Dip the macroporous resin with 2BV 5% NaOH in 80℃ water bathe for 6 hours, such as the fist method.Result:Preteat the macroporous resins AB-8 ,D-101, HPD-450, HPD-600, HPD-700 with the above four pretreatment methods .The results show that EtOH eluent volume of Vacuum evaporation method is the least , but there is a little distinction. At last, the preteat method is selected as Alkali-EtOH which is more convenience and less cost. The eluent is turbidity until its absorbtance is less than 2.240 at 254nm.The residua determination of the eluent with the different absorptance .Determining residual solvents are benzene, methyl cyclohexane, methylbenzene , ethylbenzene,o-xylene, styrene, m-xylene, divinylbenzene, naphth alene, Decane, Dodecane.The content of residua in eluent which has different absorptance (the absorptance are 2. 240 , 0.5, 0.3, 0.1, 0.03) is compared by Gas-Chromatographic. The component and content of eluent with different absorptance is compared by Gas-mass spectrometer.Result:The eluent which has different absorptance has the same component and different content. The eluent have some residua until its absorptance is 0.3. 2. Determination of the residua of the macroporous resins.2.1 comparation of the determination method to different resins Comparing the residua determination methods of different resins by theheadspace injection of the solid macroporous resin, the result indicate that the headspace injection of solid resin is a availability residua determination method.Take out AB-8, D-101, HPD-450, HPD-600, HPD-700 resin 0. 05g and determinate its residual content by solide headspace method. Study the methods of the two different standard curve which are the standard curve method of adding the standard solution into the homology pretreat resin and the standard curve method of solvent.Result indication : with the standard curve method of solvent , the recovery experiment result is low. The standard curve method of adding the standard solution into the homology pretreat resin is the feasible method with very goodly recovery. The different resin has different standard curve , its slope RSD is benenze 46.18%, methyl cyclohexane 28.19%, methyl benzene 28. 65%, ethyl benzene 20.82%, o-xylene 20.49%, styrene 21.19%, m-xylene 20.07%, the standard curve of diffenent resin is not superposition, thus, dertermination residual content should use the homology pretreat resin adding the standard solvent as comparation.2.2 Determination of the divinylbenzene of the macroporous resin.The determination method of the divinylbenzene is comparated with the direct injection and the headspace injection .The result indicate that the direct injection method of solvent is better than the headspace method because of its the lower limit detection .So that, the direct injection method is regarded as the determination method of diviylbenzene.There have some studies about the determination method which is the extracting solvents, extracting methods, extactin time, the sample volumn. Result indicate that the best method is 0. lg resin circumluence extracting 30 min with 25ml EtOH.Take out AB-8, D-101, HPD-450, HPD-600, HPD-700 resin and study the detection method of the standard curve, the precision, the respectively and the recoveryof diviylbenzen.Reslt:The linear regression equation and the linear rangs of m-diviylbenzene is Y=7. 8631X-0. 06805 (r=0. 9999) between 0.1628—50. 8821ng, p-diviylbenzene Y=8311. 932X+1. 969851 (r=0. 9999) between 0.1628—50. 8821ng, the precision show that RSD of m-diviylbenzene is 3. 9%, RSD of p-diviylbenzene is 0. 7%. stablization research indicate that AB-8 content of nrdiviylbenezene is 490.19X10~*g. g , RSD is 4.0%, the content of p-diviylbenezene is 735.10X10^g. g"1, RSD is 7.7 %.the reproducibility result indicate that AB-8 content of m-diviylbenezene is 544.15X lO^g. g , RSD is 1.8%, the content of p-diviylbenezene is 718. 44X10"^?. g , RSD is 3.7%, the content of diviylbenezene are 0 in the D-101,HPD-450,HPD-600,HPD-700. the recovery of m-diviylbenzen of AB-8, D-101,HPD-450,HPD-600,HPD-700 are 85. 6% ,89. 7%,93. 7%,93. 25%,94. 38%respectly, RSD are 4.8%, 3.1%, 2.7%, 2.4%, 2. 6%0 the recovery of p-diviylbenzen of AB-8> D-101, HPD-450, HPD-600, HPD-700 are 94.16%, 91.68%, 93.38%, 92.60%, 88.65%, RSD are 6. 8%, 1. 7%, 1. 7%, 1. 6%, 2. 5%. the low limit detection:m-diviylbenezene is 0. 0815mg. ml'1, the low detection content is 0. 08151ng, p-diviylbenzene :the low limit detetion is 0. 0834Hg. ml , the low detection content is 0. 0834ng. The pretreat resin has not the divylbenezene.Conclusion:This method has the lower detection which is ng level and is effective determination technology of the diviylbenezene.2.3 Determination of the naphthalene of the macroporous resin.Determinate to the content of naphthalene, decane, dodecane in macroporous resin with headspace Gas-chromatographic by solid resin headspace injection.Take AB-8 as example to study the determination method including standard curve , precision, reproducibility, recovery, limit detection, then determinate the content of naphthalene in sixteen type macroporous, such as D-101, HPD100, DM-1, D-101A, etc.Result: The content of naphthalene, decane, dodecane in AB-8 resin respectively are 2482. 31ppm, 7939. 93ppm, 5943. 57ppm, The linear rangs are 3.19-6315. 46ppm (r=0.9995), 9.00-12787. 49ppm (r=0. 9994), 8.03-13465. 59ppm (r=0.9991) .the precision are 10.1%, 8.8% and 9.9% respectively, the reproducibility result show that the RSD are 6. 4%, 1. 4%, 2. 8%. the stablization research show that RSD are 5. 0%, 5. 4%, 5. 7% respectively, the recovery experiment show that recovery are 94.1%, 99. 4% , 96. 02% and RSD are 4. 2%, 5.1%, 5. 5%. The low limit detection are 0. 78ppm, 0. 82ppm, 0. 65ppm resppectively.Conclusion :This method is rapid and effective and can be used to control the quality of resin .3 The evaluation of the pretreat method.Take the resin and pretreat until the eluent adding the same volumn water is not turbidity and the absorptance of its eluent are 0.5,0.3, 0.1,0.03 respectively. Then take out the pretrat resin respectly and determinate the residua with solid headspace GC.Result show that the pretreat resin until the absorptance of the eluent is less 0.5 has hardly no residual solvents. The pretreat criterion is certained that the absorptance of the eluent is less than 0. 3.4 The study about the pharmaceutical technology of macroporous resins.Determine the parameters of the static adsorption and the dynamic adsorption of HP-20, D-101, D-101A, HPD100, HPD-450 macroporous resin separating to glycyrrhizic acid, paeoniflorin and gardenoside in Radix glycyrrhizae, paeonia veitchii Lynvh, gardenia jasm ionides Eills. Estabish the method of the combination of static adsorption and dynamic adsorption for selecting to the suited macroporous resin and separating the effective components in Chinese tradition medicine.Determination of the static adsorption content: Dip 6.OOg resin with 50ml 0. 25g drug ml"1 solution for 1, 2, 4, 8,12h, determine the concentration of the solution before and after adsorption by HPLC.Determination of the dynamic content and elution ratio: put the 50 ml 0. 25g drug, ml"1 solution into the resin column with 6. OOg preteated resin. The adsorptive velovity is 2BV. h"1, put the adsorption solution to resin column again. Determine the concentration of the solution before and after adsorption by HPLC. Calculate the dynamic content. Elute the resin column with 4BV water until the eluent is colorless, then elute the column with 50% EtOH and collect the eluent 1BV per volumn, determine the content of the effective components and depict the elution curve and determination of the content of effective components in purified solid production.Result : The saturation content of paeoniflor on HPD-450^ HPD-100, D-101, D-101A, HP-20 are 83.77, 78.50, 86.03, 89.41, 68.83 mg. g"1 respectively by static adsorption method . The saturation content of paeoniflor are 59. 64, 70. 00, 79.59, 68.44, 54.69mg. g"1 respectively by the dynamic adsorption method .The elution ratio are 69. 53%, 78. 69%, 67. 61 %, 78. 55%, 86. 23%. The content of purified solid production are 21.18 % , 22. 61 % , 17. 30 % , 31. 73 % , 19. 73 % respectively.The saturation content of glycyrrhizic acid on HPD-450, HPD-100, D-101, D-101A, HP-20 are 54. 53, 47. 53, 81. 49, 54. 78, 53. 77mg. g"1 respectively by static adsorption method . The saturation content are 51. 57 , 39. 47, 69.10, 76. 87, 34. 96 mg. g"1 respectively by the dynamic adsorption method .The elution ratio are55. 81 % ^ 67. 25%, 51.18%, 49. 72%, 64. 78%. The content of purified solid production are 8. 63%, 13. 50%, 15. 21%, 13. 48%, 9. 82% respectively.The saturation content of gardenoside on HPD-450, HPD-100, D-101, D-101A, HP-20 are 142. 21,147. 61,136. 74,135. 38, 118. llmg. g"1 respectively by static adsorption method .The saturation content are 43.43, 76.51, 32.04, 56.29, 70. 09mg/g respectively by the dynamic adsorption method . The elution ratio are 39. 79 % ,63. 89 % ,27. 63 % ,55. 91 % ,68. 28 % .The content of purified solid production are 7. 21%, 19. 57%, 13. 92%, 13. 62%, 21. 83% respectively.Conclusion: D—101A is the better adsorbents for the purification of paeoniflorin. D — 101 is the effective adsorbents for the purification of glycyrrhizic. HPD—100, P —20 the better adsorbents for the purification of gardenoside.5 The study on transferal ratio of residua in the pharmaceutical technology of macroporous resins.Determination the transferal ratio of residua of AB-8, D-101 macroporous resins separating to glycyrrhizic acid, paeoniflorin and gardenoside in Radix glycyrrhizae,Radix paeoniae alba .gardenia jasm ionides Eills.Take the pretreat resins, the unpretreat resions and the resins adding the standard solution respectively , put them into the column , then put the the 50 ml 0. 25g drug, ml"1 solution into the resin column by dynamic adsorption. Elute the resin column with 4BV water, then elute the column with different content EtOH-Water and collect the eluent 1BV per volumn , determine the content of the residual solvents in elution.Result indicate that the transferal ratio of residua are 0 in the eluent of pretreat resin and the eluent of Radix paeoniae alba with 30%> 40% EtOH elute. The transferal ratio of methyl benzene is 11.14% in the eluent of Radix glycyrrhizae through AB-8 unpretreat resions with 50% EtOH elute and 23. 76% in the eluent of Radix glycyrrhizae through D-101 unpretreat resions with 50% EtOH elute. The transferal ratio of methyl benzene is 19.90% in the eluent of gardenia jasm ionides Eills through AB-8 unpretreat resions with 70% EtOH elute and 42.53% in the eluent of gardenia jasm ionides through D-101 unpretreat resions with 70% EtOH elute.Conclusion: The more the content of the eluting EtOH is , the more the transferal ratio of residual solvents is.It is feasible to decrease content of the residua in resin and pharmaceutical through the pretreat .The application of macroporous is safty0...