Therapeutic oligonucleotides have been one of the hottest areas in R&D of drugs in recent years.The research of analysis method has been widely carried out.Ion-pairing reverse liquid chromatography is the most commonly used method for good resolution and easy docking with MS.A review is presented here on the progress of oligonucleotides analysis with ion-pairing reverse liquid chromatography.The column, mobile phase, MS detection, mechanism of analysis will be discussed.

Objective To improve the related substances analysis method of cobamamide.And to further research light degradation characteristics of cobamamide.Methods Determined the related substances of solid and aqueous solution of cobamamide after light degradation by high performance liquid chromatography(HPLC) conditions,which is Merck Hibar C18 column (4.6 mm ×250 mm, 5μm), 0.05 mol/L KH2PO4 solution:acetonitrile (90:10) -80% acetonitrile as mobile by gradient elution and detection wavelength 260 nm.And compared their light stability.The main three kinds of light degradation impurities were determinate from LC-MS.Results Gradient elution made the light degradation impurities separate better.The results of precision and reproducibility tests increased to RSD =0.2% (n=6) and RSD =8% (n=5) from RSD =3.8% (n =6) and RSD=38%(n=5). Cobamamide solution was very sensitive to light, the preparation should be strict dark operation.Two of the light degradation impurities were adenosine and hydroxycobalamin, with the relative response factor 2.5 and 0.7.Conclusion New method is specific, durable and reproducible, which can be used for quality control of cobamamide.

Objective To establish a method for determining cinnamic aldehyde content in Xianggui Huazhuo capsules by gas chromatography-mass spectrometry (GC-MS). Methods The content of cinnamic aldehyde was determined by GC-MS. Separation was performed on a capillary column (30 m×0. 25 mm, 0. 25 μm) with HP-5 as the stationary phase. A programmed temperature was employed. The flow rate was 1 mL·min-1 with He as carrier gas, and split ratio was 50∶1. The injection volume was 1. 0 μL. Results The cinnamic aldehyde was well isolated from the other ingredients. A good linear relationships of cin-namic aldehyde in range of 0. 02-4. 00 mg·mL-1 was observed. The correlation coefficient was 0. 999 4. The average recovery of cinnamic aldehyde was 96. 2% , and RSD was less than 2. 11% . Conclusion The method is simple, accurate and suitable for determination of cinnamic aldehyde content.

Objective To establish the dissolution curves test method of Ribavirin capsules, and investigate dissolution behavior of domestic Ribavirin capsules.To provide experimental basis for generic drugs quality consistency evaluation.Methods According to the first dissolution method (basket method)stated in appendix Ⅹof Chinese Pharmacopeia(2010 edition),the rotation speed was 50 r/min with dissolution medium volume of 900 mL.The dissolution profiles of Ribavirin capsules in four different mediums( pH 1.2 hydrochloric acid solution,pH 4.5 acetic buffer,pH 6.8 phosphate buffer and water) were determined by HPLC.The determination was performed on C18 column with mobile phase consisted of 4 g/L sodium dihydrogen phosphate solution(pH adjusted to 5.0 ±0.05 using 5% sodium hydroxide solution)-acetonitrile(98:2)at the flow rate of 1.0 mL/min.The detection wavelength was 225 nm,and sample size was 10μL.Results The linear range of ribavirin was 2.5-200μg/mL(r=1).RSD of precision and stability tests were lower than 0.5%.The average recoveries were 101.3%, 100.7%, 100.2%, 100.4% in four mediums.Dissolution behavior of capsules can be more consistent and rapid dissolution in pH4.5 and pH6.8 mediums.But they were quite different in pH1.2 and water mediums, and some of their average dissolution at 15 min could not reach 85%.Conclusion This method is accurate and reliable.There is a difference between domestic Ribavirin capsules dissolution behavior, and the formulation processes have room for improvement.

Objective To analyze peroxide value and anisidine value of ethyl polyenoate soft capsules and imported drugs and evaluate the oxidative stability.Methods The analysis was carried out on a TSK gel ODS-100 V(250 mm ×4.6 mm,5μm)with methanol-water(98:2,V/V)as the mobile phase to determine the structure of the vitamin E as antioxidant.The influence on the antioxidation effect of tocopherol acetate andα-tocopherol as excipient in omega-3 polyunsaturated fatty acid ethyl ester drugs was evaluated.Results The structure of vitamin E as antioxidant in domestic drugs was acetate, while vitamin E as excipient in foreign drugs had the structure of α-tocopherol monomer.As antioxidant, the antioxidation effect of tocopherol acetate was better thanα-tocopherol.The structure of vitamin E had a direct impact on the antioxidation effect of omega-3 polyunsaturated fatty acid ethyl ester drugs.Conclusion The studies provide the basis for evaluate rationality of antioxidant in ethyl polyenoate soft capsules scientifically, which has positive significance for controlling the quality of the drug effectively.

Objective To study the distribution and properly of the transparent globules within Hensen cells (HC) of guinea -pig Corti organ .Methods The cochlear epithelial cells were isolated from 10 guinea pigs .The cells of cochlea were marked by Bodipy493/503 ,sudan III ,oil red O ,and osmium tetroxide .Results The transpar‐ent globules within the HCs of the guinea -pigs were green staining by Bodipy493/503 ,jacinth staining by Sudan III ,ruby red by oil red O .And they were black globules stripe as post -fixed in 1% osmium tetroxide .Conclusion The results indicate that the transparent globules within guinea -pigs HCs'lipid droplets by four methods .

Objective To explore the polymorphism and distribution of apolipoprotein E(ApoE)gene in patients with hyperlipidemia.Methods A total of 370 hyperlipidemia patients diagnosed at Hangzhou Ninth People's Hospital from October 2023 to June 2024 were selected.Gene polymorphism was detected in the blood samples of the patients using fluorescence polymerase chain reaction,and the distribution of gene polymorphism among different genders and age groups within this population was compared.Results ①The polymorphism distribution of the two genetic loci(ApoE 388T＞C and ApoE 526C＞T)in the studied sample population conforms to Hardy-Weinberg equilibrium,indicating that it is representative of the population;②Among the 370 patients,the distribution of the ApoE gene was as follows:E3/E3(66.49％),E3/E4(20.27％),E2/E3(8.65％),E2/E4(1.89％),E2/E2(1.35％),and E4/E4(1.35％).The proportion of the patient population carrying the E4 allele was about 1.88 times higher than that of those carrying the E2 allele;③The distribution differences of ApoE gene polymorphism were not related to the patients'gender and age(P＞0.05).Conclusion The distribution of ApoE gene polymorphism among hyperlipidemia patients at a hospital in Hangzhou is uneven and is not related to gender or age groups.Therefore,it is recommended to prioritize the detection of ApoE gene in patients before using statin medications to assess treatment effectiveness.

Objective To investigate the risk factors for hemorrhagic transformation(HT)of different subtypes of acute cerebral infarction in elderly patients.Methods A total of 415 elderly hospitalized patients aged ≥60 years with acute cerebral infarction were selected from a hospital in Changsha from January 2014 to December 2016 for the study.Among them,56 had hemorrhagic transformation(observation group)and 359 had no hemorrhage transformation(control group).The general demographic information,past medical history,physical examination and laboratory indicators,cranial CT/MRI examination,admission treatment and the National Institutes of Health Stroke Scale (NIHSS) score on admission were analyzed and compared between two different subtypes.Multivariate Logistic regression was used to analyze the influencing factors for HT of different subtypes of acute cerebral infarction in elderly patients.Results The incidence of HT was 26.61％(29/109)for largeartery atherothrombosis,47.50％ (19/40)for cardiac embolism and 3.12％ (8/257)for small-artery occlusion with a statistically significant differences among the three subtypes (x2 =77.923,P =0.000).The incidence of HT was 0％ (0/7)for unknown type,and 0％ (0/2)for other types.Multivariate logistic regression showed that female was the protective factor(OR =0.171,95％ CI:0.037-0.790),but high level of glycated haemoglobin A1c(HbA1c)and large infarct size were the risk factors(OR =5.949 and 20.619,95 ％CI:1.199-29.509 and 2.869-148.207)for HT due to largeartery atherothrombosis.A high score of NIHSS and high level of matrix metalloproteinase-9 (MMP-9)were the risk factors for HT due to cardioembolism(OR =20.254 and 91.200,95％CI:1.605-255.625 and 4.044-2056.710,respectively).A high level of homocysteine(Hcy) and large infarct size were the risk factors for HT due to small-artery occlusion(OR =10.231 and 15.251,95 ％ CI:1.202-87.108 and 3.371-68.995,respectively).Conclusions The incidence of HT due to cardioembolism is the highest.Influencing factors for HT due to different subtypes of acute cerebral infarction are different in the elderly.It is important for clinicians to know that gender,HbA1c levels and infarct size are associated with HT due to large-artery atherothrombosis,NIHSS score on admission and MMP-9 concentration associated with HT due to cardioembolism,and Hcy level and infarct size associated with HT due to small-artery occlusion.