课程名称︰分析化学丁
课程性质︰药学系 医学检验暨生物技术学系 必修
课程教师︰张焕宗
开课学院：理学院
开课系所︰化学系
考试日期（年月日）︰2014年 6月18日
考试时限（分钟）： 120分钟
是否需发放奖励金： 是
（如未明确表示，则不予发放）
试题 :
I. Multiple choices (Choose the right answer(s); each right pick is worthy 2
points; it costs you nothing for less than 5 wrong picks; 1, 2, and 3 points
for each wrong pick if total wrong picks are more than 2 but less than 6, 10,
and 15, respectively )
1. (A) Separation efficiency: CE > HPLC, (B) Mass transfer is a main source
for band broadening in CZE, (C) Injection volumes in HPLC and CE are in
the orders of μL and nL, respectively, (D) Microchip capillary
electrophoresis can not be used for the separation of PCR products, (E) CE
can not be used for the separation of single cells.
2. Common detectors used in GC are: (A) Flame ionization detector, (B) Mass
spectrometry, (C) Fluorescence, (D) Absorption, (E) Thermal conductivity
detector.
3. (A) 10 mM Phosphate solution containing β-CD is not useful for the
separation of chiral compounds, (B) The hydrophilic end of a drug towards
the cavity of CD, while the hydrophobic end towards aqueous solution during
recognition, (C) Aliphatic alkanes can be separated by CE using 10 mM
phosphate (pH 8.0) as a buffer, (D) 10-mM Phosphate solution (pH 8.0)
containing 20 mM SDS is a useful buffer for separating natural drugs by CE,
(E) A capillary wall coated with polymer molecules is useful for
separating proteins.
4. (A) EOF at pH 9.0 is smaller than that at pH 3.0 when using a noncoated
fused silica column, (B) At pH 4.5, EOF is higher in 25 mM acetate buffer
than that in 25 mM acetate buffer containing 0.1% PEO (a neutral polymer),
(C) The direction of EOF can be reversed by using a cationic surfactant,
(D) EOF mobility is larger in 10 mM phosphate buffer at pH 8.5 than that at
pH 5.0 when the silica wall is covered with Si-O(CH2)17CH3, (E) EOF
mobility increased upon decreasing temperature.
5. (A) For DNA separation, CZE is more suitable than that of CGE, (B) Proteins
can be separated by CZE, (C) Proteins migrated toward the cathode end when
the separation was conducted at pH greater than its pI value in the absence
of EOF, (D) CGE can be used to determine the molecular weight of unknown
proteins, (E) Peptides can be separated by CZE.
6. Gradient techniques: (A) Temperature programming is commonly used in HPLC,
(B) pH Gradient is useful for the separation of the metal ion by HPLC using
a cation exchanger, (C) Change in salt concentration is commonly used to
improve the separation of proteins by HPLC, (D) Change in solution polarity
is useful for the separation of neutral compounds by reversed phase HPLC,
(E) Change in temperature is useful to improve the separation of DNA in CE.
7. (A) Standard addition method is commonly used to improve quantitation of
analytes, (B) Adding internal standard is useful to correct the
instrumental variation for better quantitation, (C) A standard solution
containing a same compound of the analyte is used in an internal standard
method, (D) Standard addition method is better than internal standard
method for the analysis of complicated samples, (E) When applying a
standard addition method, the concentration of a known compound is much
higher than that of the unknown.
8. Which of the following techniques are commonly used for improved
sensitivity in CE? (A) Field amplification, (B) Change in temperature, (C)
pH Junction, (D) Change in solution polarity, (E) Change in viscosity.
II. Fill in the blanks (3 pts each):
1. A sample containing an acid, an amine, and a neutral compound was separated\
by CE at pH 6.0. Three peaks were detected at 2.0, 3.0 and 4.0 min at 20
kV, using a 30-cm capillary (the separation length is 20 cm), μEOF is
__(1)__, μep values for the cationic and anionic analytes are __(2)__ and
__(3)__, respectively.
2. The main source for peak broadening in CE is: __(4)__.
3. __(5)__ is useful for the separation of IgG in complicated samples.
4. The analyte and unknown at the same concentrations have signals of 100 and
110, respectively. When the standard at 100 nM has a signal of 80, the
concentration of the unknown at a signal of 160 is __(6)__.
5. Two analytes X and Y were separated by HPLC using a 50-cm long column with
retention times of 10 (width at the base 0.5 min) and 10.5 min (0.5 min),
respectively. If an unretained solute was detected at 1 min. Calculate the
(A) Retention factor of X: __(7)__; (B) Selectivity factor: __(8)__; (C)
resolution: __(9)__; (D) Efficiency for X: __(10)__; (E) Plate height for
X: __(11)__; The column length required for resolution 1.6: __(12)__.
6. Name the three major factors for peak broadening in chromatography:
__(13)__ __(14)__ and __(15)__. Write down the van Deemter equation:
__(16)__
III. use schemes/figures to answer the following question: (3 pts each; No
explanation)
1. CE system
2. HPLC system
3. GC system
4. Support-coated open tubular column
5. An electropherogram of the separation of isotopic chloride ions by CE with
indirect detection
6. A scheme for stacking of cations
7. A schematic setup of a separation column with a suppressor
8. A figure of log Mw vs. retention time in HPLC
9. Irregular peak shape due to sample overloading in HPLC
10. Peak height vs. flow rate
11. Effect of stationary film thickness on the plate height
12. A plot for analyte quantitation when a standard addition method is applied
13. On-line concentration of DNA in CE using PEO solution