The Research Of Quality Standard Of TFAs,MCPDEs And Sterol Composition In Fats And Oils Of Pharmaceutical Excipients

Oil is an important source of human nutrition,not only for meeting energy needs,but also meeting requirements for essential fatty acids and vitamins.Its healthcare and medicinal value has been developed and wildly used.Whether it is as edible fats or medicinal raw materials grease,the quality control of fats and oils is extremely important.For example,trans fatty acids(TFAs)which may endanger the risk of cardiovascular disease and monochloropropanol esters(MCPDEs)with certain renal toxicity and reproductive toxicity which have caused extensive concern in the field of food and been researched a lot both at home and abroad,were not recorded in the domestic and foreign pharmacopoeias or standards.And the composition and content of sterols plays an important role in the quality of vegetable oil and process evaluation,which has been contained in USP 40 and EP 9.0/BP 2017 while Chinese Pharmacopoeia has not yet received.Trans Fatty Acids In this study,the conditions of sample pretreatment(Ag+-SPE pre-separation method)and chromatographic separations of GC and GC-MS were optimized.The detection method for TFAs in fats and oils of pharmaceutical excipients was established,so as to improve the quality standard of pharmaceutical excipients.According to the internal standard calibration,an excellent linear correlation(r2>0.997)in a certain concentration range was obtained.The method was used to detected FAPAS QC test material and the z-score was less than two,which verified the feasibility and accuracy of the method.The method was validated to be sensitive,precise and accurate.The standard method of quantitative detection of TFAs in the fats and oils of pharmaceutical excipients was developed.Seven different fats and oils including soybean oils(for pharmaceutic adjuvant and injection),corn oils,peanut oils,olive oils,coconut oils,hydrogenated soybean oils and hydrogenated castor oil were chosen as samples.The oil samples were methylated to fatty acids methyl esters using BF3 in methanol.TFAs were quantitatively measured by GC-MS against C11:0 internal standard.Fifty-two fatty acid methyl esters could be well separated under the experiment conditions.The total content of TFAs of the pharmaceutical excipients soybean oil was the highest(>2%),followed by the refined corn oil(?1.5%),and the total amount of olive oil and peanut oil was the least(<1%).According to the limit value,the total content of TFAs in the medicinal fats should not be over 1.22%(strictly),so it was necessary to control the TFAs in the fats and oils of pharmaceutical excipients,especially in soybean oil for pharmaceutic excipients.In this study,a GC method and a GC-MS method were investigated.The two methods had their advantages.The GC method was more conventional,common and low cost.While the GC-MS method had high sensitivity,and the MS detector can provide the spectrum,combined with the advantages of GC,which makes the results more accurate and reliable.Monochloropropanol Esters A method for the quantitative determination of 3-monochloropropane-1,2-diol esters(3-MCPDEs)and 2-monochloropropane-1,3-diol esters(2-MCPDEs)in oils was presented using GC-MS coupled with the isotopic internal standard(deuterium)technique.The optimum conditions for the sample pretreatment were investigated.MCPD was released from MCPDEs by transesterification with NaOCH3/methanol.After derivatization with phenylboronic acid,derivative of MCPD was determined by GC-MS.According to the internal standard calibration,there was an excellent linear correlation(r2>0.999)in the range of 20-2000 ng of MCPDEs(shown as MCPD)was obtained.The method was used to detected FAPAS QC test material and the z-score was less than two,which verified the feasibility and accuracy of the method.The result showed that this method was suitable for the detection of MCPDEs in oil.The MCPDEs content of 11 kinds of oil samples including soybean oil,corn oil,peanut oil,olive oil and so on were tested.The results showed that the content of MCPDEs in oils could be divided into three grades from low to high:(1)< 1 mg / kg,such as medicinal peanut oil,soybean oil,sunflower oil,coconut oil,olive oil,refined palm kernel oil and corn oil;(2)1 ~ 3 mg / kg,such as finished fraction of refined palm stearin,rice oil and fish oil;(3)> 3 mg / kg,such as palm oil,RBD palm oil and edible peanut oil.Sterol Composition In this study,the composition and content of sterols in the 15 oil samples were analyzed.Unsaponifiable matters were extracted from olive oil,and unsaponified sterol fractions were separated and collected by HPLC,then the sterol fractions were derived before analysed by GC to obtain the composition and content of sterol.As basic data,the contents of fifteen kinds of sterol components were used by cluster analysis with SPSS 18.0 to study the difference of sterol spectrum in different oils and evaluate the consistency of the process.The results showed that the oils analyzed contained about 15 kinds of sterols.In these oils,expert lecithin and palm kernel oil,the content of ?-sitosterol are higher,followed by campesterol and ?5,24-stigmastadienol.The main composition of the sterols in lecithin is cholesterol,accounting for more than 97%,and it is ?5-avenasterol in palm kernel oil,accounting for about 70%.The cluster analysis can be used to initially distinguish oil species,and the consistency of oil process can be judged based on the clustering results.Based on the percentage of 15 kinds of sterols,the cluster analysis can be used to distinguish the oil species.According to the clustering results,the consistency of the oil process could be judged initially.For example,soybean oil of the same level and the same manufacturer can be divided into one class,indicating that the same process.Different processes lead to differences in the composition of sterols in soybean oil of the same manufacturers and different use.There were sterol composition differences between several batches of almond oil of the same manufacturers,which could be used to infer the process differences between these batches of almond oil.As a result,the determination of TFAs and MCPDEs in fats and oils of pharmaceutical excipients was established in my work.The composition of sterols in oil was analyzed,and the differences of sterol composition as well as the consistency of process were analyzed by SPSS respectively.This study provides a basis for the establishment of the relevant testing standards and the collection of pharmacopoeia.