Yang Rui-fen, Shi Zhi-guo, Feng Yu-qi, Da Shi-lu
Hubei College of Pharmacy and Medical Laboratory, Wuhan 430064, China.
Yao Xue Xue Bao. 2003 Feb;38(2):129-32.
To establish separation methods of five sulfonamides by using capillary high performance liquid chromatography(mu-HPLC) and electrochromatography. The effect of mobile phase varies such as methanol content, pH, buffer solution concentration and voltage on their chromatographic behavior and electroosmesis flow was investigated. Capillary electrochromatography (CEC) was compared with mu-HPLC at the same condition.
Stationary phase was ODS, mobile phase was methanol and 2 mmol.L-1 H3PO4 buffer solution (pH 3.0-7.0), voltage was 0- -15 kV, flow rate was 10 microL.min-1, pressure was approximately 70 MPa and UV detection wavelength was 254 nm.
Separations on base line have been respectively accomplished for five sulfonamides by mu-HPLC with mobile phase of methanol-2 mmol.L-1 H3PO4 buffer solution (30:70) at pH 5.0 in 67 min, and CEC with the same mobile phase at -5 kV voltage in 25 min.
Electroosmesis flow of CEC decreased with the increase in methanol content, buffer solution concentration, increased with the increase in voltage and increase slightly with the increase in pH of mobile phase. Retention values (k) of solutes to be examined decreased with increasing methanol content of mobile phase in mu-HPLC and CEC. Retention values (k) of solutes increased slightly with increasing buffer solution concentration, decreased with increasing voltage in CEC. Trimethoprim(TMP) decreased obviously with increasing voltage in CEC. The effect of pH of mobile phase on retention values (k) was more complex. Five sulfonamides were separated at the same mobile phase condition by mu-HPLC and CEC. And separation speed of CEC was much faster than that of mu-HPLC. CEC was very fit for rapid separation of sulfonamides.
建立5种磺胺类药物的毛细管高效液相色谱（μ-HPLC）和毛细管电色谱分离方法。考察了流动相组成如甲醇含量、pH值、缓冲溶液浓度及电压等对其色谱行为和电渗流的影响，并在相同条件下比较了毛细管电色谱（CEC）与μ-HPLC。
固定相为ODS，流动相为甲醇-2 mmol·L-1 H3PO4缓冲溶液（pH 3.0～7.0），电压为0～-15 kV，流速为10 μL·min-1，压力约为70 MPa，紫外检测波长为254 nm。
μ-HPLC采用甲醇-2 mmol·L-1 H3PO4缓冲溶液（30:70）为流动相，pH 5.0，67 min内可基线分离5种磺胺类药物；CEC采用相同流动相，-5 kV电压，25 min内可完成分离。
CEC的电渗流随甲醇含量、缓冲溶液浓度的增加而降低，随电压的增加而增加，随流动相pH值的增加略有增加。μ-HPLC和CEC中，待检测溶质的保留值（k）均随流动相甲醇含量的增加而降低。CEC中，溶质的保留值（k）随缓冲溶液浓度的增加略有增加，随电压的增加而降低。甲氧苄啶（TMP）在CEC中随电压增加下降明显。流动相pH值对保留值（k）的影响较复杂。μ-HPLC和CEC在相同流动相条件下均可分离5种磺胺类药物，且CEC的分离速度远快于μ-HPLC。CEC非常适合磺胺类药物的快速分离。
