高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (9): 1893.doi: 10.7503/cjcu20180262
收稿日期:
2018-04-04
出版日期:
2018-09-07
发布日期:
2018-06-26
作者简介:
联系人简介: 练鸿振, 男, 博士, 教授, 博士生导师, 主要从事色谱分离和分析方面的研究. E-mail:
基金资助:
QIAO Junqin, LIANG Chao, CAO Zhaoming, LIAN Hongzhen*()
Received:
2018-04-04
Online:
2018-09-07
Published:
2018-06-26
Contact:
LIAN Hongzhen
E-mail:hzlian@nju.edu.cn
Supported by:
摘要:
利用离子对反相液相色谱(IP-RPLC)对寡核苷酸在混合离子对试剂三乙胺/丙胺-乙酸盐(TEA/PA-AA)体系下的保留行为进行了研究, 并与经典离子对试剂三乙胺乙酸盐(TEAA)体系下的保留行为进行了对比. 实验结果表明, 相同离子对试剂浓度下, 寡核苷酸在TEA/PA-AA体系下的保留均弱于TEAA体系下的保留, 且寡核苷酸的保留均随着离子对试剂浓度(20~120 mmol/L)的增加而增强. 同型寡核苷酸(dC)n作为特例, 当n>10时, 保留基本趋于稳定, 这是由于(dC)n随着离子对试剂浓度的增加保留增长较快, 在较低的离子对试剂浓度下即可达到最大保留. 同时发现, 短链同型寡核苷酸(dT)n和异型寡核苷酸在TEA/PA-AA体系下的分离均优于TEAA体系, 而同型寡核苷酸(dA)n和(dC)n的分离则在TEAA体系下更优. 通过研究流动相中离子对试剂总浓度cp与寡核苷酸保留因子k之间的关系, 推断出2种体系下寡核苷酸的保留机理均以离子对模型占主导地位. 总体而言, 分离相同长度的异型寡核苷酸时, TEA/PA-AA混合离子对体系尤其在中等离子对浓度下比TEAA体系具有明显优势, 低的离子对试剂浓度可增加与后续电喷雾质谱(ESI-MS)的兼容性, 有利于寡核苷酸的定性分析.
中图分类号:
TrendMD:
乔俊琴, 梁超, 曹兆明, 练鸿振. 混合离子对试剂体系下寡核苷酸的IP-RPLC保留行为. 高等学校化学学报, 2018, 39(9): 1893.
QIAO Junqin,LIANG Chao,CAO Zhaoming,LIAN Hongzhen. Retention Behavior of Oligonucleotides under System Containing Mixed Ion-pair Reagents by IP-RPLC†. Chem. J. Chinese Universities, 2018, 39(9): 1893.
Oligonucleotide | Sequence(5'-3') | Percentage(%) | ΔG*/(J·mol-1) | |||
---|---|---|---|---|---|---|
G | C | A | T | |||
Oligo20-1 | CTTAGTGAAGAGCTCAGTTA | 25 | 15 | 30 | 30 | -3.97×104 |
Oligo20-2 | CTTAGTGAAGAGTCTCTAAG | 25 | 15 | 30 | 30 | -2.55×104 |
Oligo20-3 | GACAGGAAAGACATTCTGGC | 30 | 20 | 35 | 15 | -1.48×104 |
Oligo20-4 | GACAGGAAAGACATTCCGGT | 30 | 20 | 35 | 15 | -4.08×104 |
Oligo32-1 | GTCGTTATCATCAGAGTAGCCCAGGAAGCTTC | 25 | 25 | 25 | 25 | -5.60×104 |
Oligo32-2 | GCGTACAGTATAGCCCAGTCTTGAGTGCCATA | 25 | 25 | 25 | 25 | -1.53×104 |
Oligo32-3 | GGTATGGTTTACGAGTATTGCCTGAAGCGAGG | 37.5 | 12.5 | 21.9 | 28.1 | -1.95×104 |
Oligo32-4 | CCTCGCTTCAGGCAATACTCGTAAACCATACC | 12.5 | 37.5 | 28.1 | 21.9 | -1.95×104 |
Table 1 Hetero-oligonucleotides used in the experiment
Oligonucleotide | Sequence(5'-3') | Percentage(%) | ΔG*/(J·mol-1) | |||
---|---|---|---|---|---|---|
G | C | A | T | |||
Oligo20-1 | CTTAGTGAAGAGCTCAGTTA | 25 | 15 | 30 | 30 | -3.97×104 |
Oligo20-2 | CTTAGTGAAGAGTCTCTAAG | 25 | 15 | 30 | 30 | -2.55×104 |
Oligo20-3 | GACAGGAAAGACATTCTGGC | 30 | 20 | 35 | 15 | -1.48×104 |
Oligo20-4 | GACAGGAAAGACATTCCGGT | 30 | 20 | 35 | 15 | -4.08×104 |
Oligo32-1 | GTCGTTATCATCAGAGTAGCCCAGGAAGCTTC | 25 | 25 | 25 | 25 | -5.60×104 |
Oligo32-2 | GCGTACAGTATAGCCCAGTCTTGAGTGCCATA | 25 | 25 | 25 | 25 | -1.53×104 |
Oligo32-3 | GGTATGGTTTACGAGTATTGCCTGAAGCGAGG | 37.5 | 12.5 | 21.9 | 28.1 | -1.95×104 |
Oligo32-4 | CCTCGCTTCAGGCAATACTCGTAAACCATACC | 12.5 | 37.5 | 28.1 | 21.9 | -1.95×104 |
Fig.1 Chromatograms of (dA)n under mobile phases containing either 10 mmol/L TEA/10 mmol/L PA(A)or 20 mmol/L TEA(B)Purospher© STAR RP-18 endcapped column(50 mm×4.6 mm i.d., 5 μm); column temperature: 30 ℃; mobile phase A(10 mmol/L TEA/10 mmol/L PA-20 mmol/LAA-5%CH3CN, pH=7.0) or (20 mmol/L TEAA-5%CH3CN, pH=7.0), mobile phase B(10 mmol/L TEA/10 mmol/L PA-20 mmol/L AA-25%CH3CN, pH=7.0) or (20 mmol/L TEAA-25%CH3CN, pH=7.0); gradient elution: 0—40 min, 15%B—45%B; 40—43 min, 45%B—15%B; 43—58 min, 15%B—15%B; flow rate: 1.0 mL/min; detection wavelength: 260 nm.
Fig.2 Retention factor k of (dA)n under mobile phases containing different concentrations of ion-pair reagent(A) TEA/PA; (B) TEA. The concentrations of ion-pair reagent in mobile phase were 20, 40, 60, 80, 100 and 120 mmol/L, respectively. Other chromatographic conditions were the same as in Fig.1.
TEA/PA | TEA | |||||
---|---|---|---|---|---|---|
A10/A20 | A20/A25 | A25/A30 | A10/A20 | A20/A25 | A25/A30 | |
20 | 5.02 | 1.76 | 1.34 | 9.06 | 2.73 | 1.97 |
40 | 9.20 | 2.66 | 1.89 | 12.47 | 3.47 | 2.55 |
60 | 10.90 | 2.96 | 2.12 | 13.36 | 3.74 | 2.77 |
80 | 11.58 | 3.17 | 2.29 | 13.98 | 3.92 | 2.89 |
100 | 12.37 | 3.33 | 2.37 | 14.76 | 4.03 | 2.96 |
120 | 12.55 | 3.34 | 2.39 | 14.42 | 3.99 | 2.94 |
Table 2 Resolution(Rs) of adjacent peaks in (dA)n under different concentrations of ion-pair reagent
TEA/PA | TEA | |||||
---|---|---|---|---|---|---|
A10/A20 | A20/A25 | A25/A30 | A10/A20 | A20/A25 | A25/A30 | |
20 | 5.02 | 1.76 | 1.34 | 9.06 | 2.73 | 1.97 |
40 | 9.20 | 2.66 | 1.89 | 12.47 | 3.47 | 2.55 |
60 | 10.90 | 2.96 | 2.12 | 13.36 | 3.74 | 2.77 |
80 | 11.58 | 3.17 | 2.29 | 13.98 | 3.92 | 2.89 |
100 | 12.37 | 3.33 | 2.37 | 14.76 | 4.03 | 2.96 |
120 | 12.55 | 3.34 | 2.39 | 14.42 | 3.99 | 2.94 |
Fig.3 Retention factor k of (dT)n under mobile phases containing different concentrations of ion-pair reagent (A) TEA/PA; (B) TEA. The chromatographic conditions are the same as those in Fig.2.
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
T5/T7 | T7/T10 | T10/T12 | T5/T7 | T7/T10 | T10/T12 | |
20 | 10.28 | 9.24 | 3.78 | 9.79 | 9.26 | 4.08 |
40 | 11.79 | 10.42 | 4.44 | 10.96 | 10.20 | 4.53 |
60 | 12.50 | 10.96 | 4.76 | 11.58 | 10.48 | 4.85 |
80 | 12.92 | 11.32 | 4.85 | 11.83 | 10.98 | 5.06 |
100 | 13.32 | 11.63 | 5.06 | 12.24 | 11.40 | 5.07 |
120 | 13.49 | 11.78 | 5.31 | 12.54 | 11.60 | 5.26 |
Table 3 Resolution(Rs) of adjacent peaks in (dT)n under different concentrations of ion-pair reagent
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
T5/T7 | T7/T10 | T10/T12 | T5/T7 | T7/T10 | T10/T12 | |
20 | 10.28 | 9.24 | 3.78 | 9.79 | 9.26 | 4.08 |
40 | 11.79 | 10.42 | 4.44 | 10.96 | 10.20 | 4.53 |
60 | 12.50 | 10.96 | 4.76 | 11.58 | 10.48 | 4.85 |
80 | 12.92 | 11.32 | 4.85 | 11.83 | 10.98 | 5.06 |
100 | 13.32 | 11.63 | 5.06 | 12.24 | 11.40 | 5.07 |
120 | 13.49 | 11.78 | 5.31 | 12.54 | 11.60 | 5.26 |
Fig.4 Retention factor(k) of (dC)n under mobile phases containing different concentrations of ion-pair reagent (A), (C) TEA/PA; (B), (D) TEA. The chromatographic conditions are the same as those in Fig.2.
System | cp/(mmol·L-1) | C5/C6 | C6/C7 | C7/C8 | C8/C9 | C10/C15 | C15/C20 | C20/C25 |
---|---|---|---|---|---|---|---|---|
TEA/PA | 20 | 0.43 | 0.76 | 1.15 | 1.53 | 12.90 | 5.24 | 1.88 |
40 | 1.21 | 1.33 | 1.38 | 1.68 | 12.03 | 5.44 | 2.09 | |
60 | 1.54 | 1.58 | 1.80 | 2.02 | 11.46 | 5.37 | 2.01 | |
80 | 1.67 | 1.81 | 1.91 | 2.12 | 11.37 | 5.47 | 1.93 | |
100 | 1.72 | 1.86 | 1.94 | 2.10 | 11.43 | 5.09 | 2.06 | |
120 | 1.79 | 1.95 | 2.02 | 2.16 | 10.94 | 5.45 | 2.15 | |
TEA | 20 | 1.16 | 1.25 | 1.67 | 2.00 | 13.01 | 5.57 | 2.00 |
40 | 1.69 | 1.86 | 1.98 | 2.35 | 13.46 | 5.53 | 1.94 | |
60 | 1.82 | 1.98 | 2.12 | 2.46 | 13.05 | 5.51 | 1.97 | |
80 | 1.91 | 1.98 | 2.16 | 2.41 | 12.48 | 5.62 | 1.99 | |
100 | 1.86 | 1.93 | 2.07 | 2.29 | 12.36 | 5.31 | 2.01 | |
120 | 1.73 | 1.90 | 1.96 | 2.15 | 12.04 | 5.71 | 2.04 |
Table 4 Resolution(Rs) of adjacent peaks in (dC)n under different concentrations of ion-pair reagent
System | cp/(mmol·L-1) | C5/C6 | C6/C7 | C7/C8 | C8/C9 | C10/C15 | C15/C20 | C20/C25 |
---|---|---|---|---|---|---|---|---|
TEA/PA | 20 | 0.43 | 0.76 | 1.15 | 1.53 | 12.90 | 5.24 | 1.88 |
40 | 1.21 | 1.33 | 1.38 | 1.68 | 12.03 | 5.44 | 2.09 | |
60 | 1.54 | 1.58 | 1.80 | 2.02 | 11.46 | 5.37 | 2.01 | |
80 | 1.67 | 1.81 | 1.91 | 2.12 | 11.37 | 5.47 | 1.93 | |
100 | 1.72 | 1.86 | 1.94 | 2.10 | 11.43 | 5.09 | 2.06 | |
120 | 1.79 | 1.95 | 2.02 | 2.16 | 10.94 | 5.45 | 2.15 | |
TEA | 20 | 1.16 | 1.25 | 1.67 | 2.00 | 13.01 | 5.57 | 2.00 |
40 | 1.69 | 1.86 | 1.98 | 2.35 | 13.46 | 5.53 | 1.94 | |
60 | 1.82 | 1.98 | 2.12 | 2.46 | 13.05 | 5.51 | 1.97 | |
80 | 1.91 | 1.98 | 2.16 | 2.41 | 12.48 | 5.62 | 1.99 | |
100 | 1.86 | 1.93 | 2.07 | 2.29 | 12.36 | 5.31 | 2.01 | |
120 | 1.73 | 1.90 | 1.96 | 2.15 | 12.04 | 5.71 | 2.04 |
Fig.5 Retention factor(k) of 20mer hetero-oligonucleotides under mobile phases containing different concentrations of ion-pair reagent (A) TEA/PA; (B) TEA. The chromatographic conditions are the same as those in Fig.2.
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
20-4/20-3 | 20-3/20-2 | 20-2/20-1 | 20-4/20-3 | 20-3/20-2 | 20-2/20-1 | |
20 | 2.18 | 4.41 | 0.87 | 1.50 | 3.90 | 1.29 |
40 | 3.34 | 6.29 | 0.61 | 1.61 | 5.00 | 1.32 |
60 | 3.73 | 6.88 | 0.61 | 1.88 | 5.88 | 1.15 |
80 | 4.01 | 7.59 | 0.28 | 2.12 | 6.65 | 0.84 |
100 | 4.39 | 8.52a | 0.02b | 2.42 | 7.29 | 0.60 |
120 | 4.53 | 8.74a | 0.34b | 2.65 | 7.84 | 0.28 |
Table 5 Resolution(Rs) of adjacent peaks in 20mer hetero-oligonucleotides under different concentrations of ion-pair reagent
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
20-4/20-3 | 20-3/20-2 | 20-2/20-1 | 20-4/20-3 | 20-3/20-2 | 20-2/20-1 | |
20 | 2.18 | 4.41 | 0.87 | 1.50 | 3.90 | 1.29 |
40 | 3.34 | 6.29 | 0.61 | 1.61 | 5.00 | 1.32 |
60 | 3.73 | 6.88 | 0.61 | 1.88 | 5.88 | 1.15 |
80 | 4.01 | 7.59 | 0.28 | 2.12 | 6.65 | 0.84 |
100 | 4.39 | 8.52a | 0.02b | 2.42 | 7.29 | 0.60 |
120 | 4.53 | 8.74a | 0.34b | 2.65 | 7.84 | 0.28 |
Fig.6 Retention factor(k) of 32mer hetero-oligonucleotides under mobile phases containing different concentrations of ion-pair reagent (A) TEA/PA; (B) TEA. The chromatographic conditions a the same as those in Fig.2.
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
32-1/32-2 | 32-2/32-3 | 32-3/32-4 | 32-1/32-2 | 32-2/32-3 | 32-3/32-4 | |
20 | 0.49 | 0.02 | 4.92 | 0.10a | 0.02b | 5.07c |
40 | 0.48 | 0.13 | 5.24 | 0.01 | 0.09 | 4.22 |
60 | 0.46 | 0.07 | 5.16 | 0.06 | 0.04 | 3.84 |
80 | 0.48 | 0.10 | 4.81 | 0.11 | 0.01 | 3.65 |
100 | 0.50 | 0.10 | 4.65 | 0.10 | 0.05 | 3.61 |
120 | 0.50 | 0.16 | 4.48 | 0.09 | 0.03 | 3.42 |
Table 6 Resolution of adjacent peaks in 32mer hetero-oligonucleotides under different concentrations of ion-pair reagent
cp/(mmol·L-1) | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
32-1/32-2 | 32-2/32-3 | 32-3/32-4 | 32-1/32-2 | 32-2/32-3 | 32-3/32-4 | |
20 | 0.49 | 0.02 | 4.92 | 0.10a | 0.02b | 5.07c |
40 | 0.48 | 0.13 | 5.24 | 0.01 | 0.09 | 4.22 |
60 | 0.46 | 0.07 | 5.16 | 0.06 | 0.04 | 3.84 |
80 | 0.48 | 0.10 | 4.81 | 0.11 | 0.01 | 3.65 |
100 | 0.50 | 0.10 | 4.65 | 0.10 | 0.05 | 3.61 |
120 | 0.50 | 0.16 | 4.48 | 0.09 | 0.03 | 3.42 |
Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
Intercept | Slope | R2 | Intercept | Slope | R2 | |
A10 | 1.6110 | 0.0310 | 0.841 | 2.9782 | 0.0266 | 0.816 |
A20 | 2.7694 | 0.0452 | 0.830 | 5.0068 | 0.0376 | 0.806 |
A25 | 3.2356 | 0.0480 | 0.830 | 5.7163 | 0.0399 | 0.804 |
A30 | 3.6109 | 0.0498 | 0.831 | 6.2791 | 0.0417 | 0.802 |
T5 | 6.9595 | 0.0204 | 0.834 | 7.6807 | 0.0165 | 0.894 |
T7 | 8.8489 | 0.0268 | 0.849 | 9.4400 | 0.0225 | 0.911 |
T10 | 10.6040 | 0.0314 | 0.866 | 11.1440 | 0.0274 | 0.923 |
T12 | 11.3850 | 0.0333 | 0.868 | 11.9050 | 0.0297 | 0.929 |
C5 | 0.2155 | 0.0027 | 0.772 | 0.4297 | 0.0017 | 0.496 |
C6 | 0.2714 | 0.0039 | 0.783 | 0.5674 | 0.0025 | 0.488 |
C7 | 0.3539 | 0.0054 | 0.808 | 0.7494 | 0.0034 | 0.459 |
C8 | 0.4859 | 0.0071 | 0.815 | 1.0169 | 0.0041 | 0.397 |
C9 | 0.6992 | 0.0088 | 0.816 | 1.4032 | 0.0046 | 0.316 |
Oligo20-1 | 5.0833 | 0.0396 | 0.860 | 6.3444 | 0.0330 | 0.867 |
Oligo20-2 | 4.8244 | 0.0422 | 0.880 | 5.9811 | 0.0353 | 0.893 |
Oligo20-3 | 3.9152 | 0.0334 | 0.836 | 5.2573 | 0.0266 | 0.836 |
Oligo20-4 | 3.4328 | 0.0289 | 0.814 | 4.9940 | 0.0239 | 0.788 |
Oligo32-1 | 6.6123 | 0.0386 | 0.860 | 7.9326 | 0.0319 | 0.876 |
Oligo32-2 | 6.7599 | 0.0383 | 0.864 | 7.9301 | 0.0322 | 0.879 |
Oligo32-3 | 6.7601 | 0.0386 | 0.863 | 7.9182 | 0.0324 | 0.870 |
Oligo32-4 | 8.4744 | 0.0331 | 0.852 | 9.3990 | 0.02617 | 0.915 |
Table 7 Linear correlation between k and cp
Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
Intercept | Slope | R2 | Intercept | Slope | R2 | |
A10 | 1.6110 | 0.0310 | 0.841 | 2.9782 | 0.0266 | 0.816 |
A20 | 2.7694 | 0.0452 | 0.830 | 5.0068 | 0.0376 | 0.806 |
A25 | 3.2356 | 0.0480 | 0.830 | 5.7163 | 0.0399 | 0.804 |
A30 | 3.6109 | 0.0498 | 0.831 | 6.2791 | 0.0417 | 0.802 |
T5 | 6.9595 | 0.0204 | 0.834 | 7.6807 | 0.0165 | 0.894 |
T7 | 8.8489 | 0.0268 | 0.849 | 9.4400 | 0.0225 | 0.911 |
T10 | 10.6040 | 0.0314 | 0.866 | 11.1440 | 0.0274 | 0.923 |
T12 | 11.3850 | 0.0333 | 0.868 | 11.9050 | 0.0297 | 0.929 |
C5 | 0.2155 | 0.0027 | 0.772 | 0.4297 | 0.0017 | 0.496 |
C6 | 0.2714 | 0.0039 | 0.783 | 0.5674 | 0.0025 | 0.488 |
C7 | 0.3539 | 0.0054 | 0.808 | 0.7494 | 0.0034 | 0.459 |
C8 | 0.4859 | 0.0071 | 0.815 | 1.0169 | 0.0041 | 0.397 |
C9 | 0.6992 | 0.0088 | 0.816 | 1.4032 | 0.0046 | 0.316 |
Oligo20-1 | 5.0833 | 0.0396 | 0.860 | 6.3444 | 0.0330 | 0.867 |
Oligo20-2 | 4.8244 | 0.0422 | 0.880 | 5.9811 | 0.0353 | 0.893 |
Oligo20-3 | 3.9152 | 0.0334 | 0.836 | 5.2573 | 0.0266 | 0.836 |
Oligo20-4 | 3.4328 | 0.0289 | 0.814 | 4.9940 | 0.0239 | 0.788 |
Oligo32-1 | 6.6123 | 0.0386 | 0.860 | 7.9326 | 0.0319 | 0.876 |
Oligo32-2 | 6.7599 | 0.0383 | 0.864 | 7.9301 | 0.0322 | 0.879 |
Oligo32-3 | 6.7601 | 0.0386 | 0.863 | 7.9182 | 0.0324 | 0.870 |
Oligo32-4 | 8.4744 | 0.0331 | 0.852 | 9.3990 | 0.02617 | 0.915 |
Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
A1 | B1 | R2 | A1 | B1 | R2 | |
A10 | 0.1012 | 10.026 | 0.984 | 0.1339 | 4.1449 | 0.991 |
A20 | 0.0750 | 5.6674 | 0.985 | 0.0907 | 2.2607 | 0.993 |
A25 | 0.0737 | 4.6767 | 0.988 | 0.0834 | 1.9013 | 0.994 |
A30 | 0.0726 | 4.0594 | 0.990 | 0.0784 | 1.6810 | 0.994 |
T5 | 0.1035 | 0.8199 | 0.987 | 0.1024 | 0.5507 | 0.957 |
T7 | 0.0791 | 0.7760 | 0.999 | 0.0815 | 0.4808 | 0.955 |
T10 | 0.0677 | 0.5527 | 0.987 | 0.0686 | 0.4111 | 0.948 |
T12 | 0.0636 | 0.4860 | 0.980 | 0.0640 | 0.3872 | 0.946 |
C5 | 1.3103 | 69.715 | 0.984 | 1.4721 | 20.595 | 0.942 |
C6 | 0.8572 | 58.660 | 0.981 | 1.0707 | 16.734 | 0.937 |
C7 | 0.6338 | 44.098 | 0.986 | 0.7867 | 13.573 | 0.930 |
C8 | 0.5083 | 30.667 | 0.990 | 0.6100 | 9.5254 | 0.915 |
C9 | 0.4201 | 19.968 | 0.993 | 0.4803 | 6.1408 | 0.877 |
C10 | 0.3614 | 11.452 | 0.994 | 0.3818 | 3.6390 | 0.762 |
Oligo20-1 | 0.0901 | 2.0868 | 0.998 | 0.0825 | 1.3597 | 0.995 |
Oligo20-2 | 0.0881 | 2.2844 | 0.998 | 0.0821 | 1.2991 | 0.993 |
Oligo20-3 | 0.1079 | 2.9454 | 0.999 | 0.0881 | 2.2844 | 0.998 |
Oligo20-4 | 0.1228 | 3.4268 | 0.997 | 0.1079 | 2.9454 | 0.999 |
Oligo32-1 | 0.0825 | 1.3597 | 0.995 | 0.0818 | 0.8813 | 0.989 |
Oligo32-2 | 0.0821 | 1.2991 | 0.993 | 0.0816 | 0.8847 | 0.989 |
Oligo32-3 | 0.0817 | 1.3088 | 0.994 | 0.0813 | 0.8989 | 0.991 |
Oligo32-4 | 0.0773 | 0.8182 | 0.988 | 0.0786 | 0.5428 | 0.960 |
Table 8 Correlations of k-1-cp fitted according to Eq.(9)
Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|
A1 | B1 | R2 | A1 | B1 | R2 | |
A10 | 0.1012 | 10.026 | 0.984 | 0.1339 | 4.1449 | 0.991 |
A20 | 0.0750 | 5.6674 | 0.985 | 0.0907 | 2.2607 | 0.993 |
A25 | 0.0737 | 4.6767 | 0.988 | 0.0834 | 1.9013 | 0.994 |
A30 | 0.0726 | 4.0594 | 0.990 | 0.0784 | 1.6810 | 0.994 |
T5 | 0.1035 | 0.8199 | 0.987 | 0.1024 | 0.5507 | 0.957 |
T7 | 0.0791 | 0.7760 | 0.999 | 0.0815 | 0.4808 | 0.955 |
T10 | 0.0677 | 0.5527 | 0.987 | 0.0686 | 0.4111 | 0.948 |
T12 | 0.0636 | 0.4860 | 0.980 | 0.0640 | 0.3872 | 0.946 |
C5 | 1.3103 | 69.715 | 0.984 | 1.4721 | 20.595 | 0.942 |
C6 | 0.8572 | 58.660 | 0.981 | 1.0707 | 16.734 | 0.937 |
C7 | 0.6338 | 44.098 | 0.986 | 0.7867 | 13.573 | 0.930 |
C8 | 0.5083 | 30.667 | 0.990 | 0.6100 | 9.5254 | 0.915 |
C9 | 0.4201 | 19.968 | 0.993 | 0.4803 | 6.1408 | 0.877 |
C10 | 0.3614 | 11.452 | 0.994 | 0.3818 | 3.6390 | 0.762 |
Oligo20-1 | 0.0901 | 2.0868 | 0.998 | 0.0825 | 1.3597 | 0.995 |
Oligo20-2 | 0.0881 | 2.2844 | 0.998 | 0.0821 | 1.2991 | 0.993 |
Oligo20-3 | 0.1079 | 2.9454 | 0.999 | 0.0881 | 2.2844 | 0.998 |
Oligo20-4 | 0.1228 | 3.4268 | 0.997 | 0.1079 | 2.9454 | 0.999 |
Oligo32-1 | 0.0825 | 1.3597 | 0.995 | 0.0818 | 0.8813 | 0.989 |
Oligo32-2 | 0.0821 | 1.2991 | 0.993 | 0.0816 | 0.8847 | 0.989 |
Oligo32-3 | 0.0817 | 1.3088 | 0.994 | 0.0813 | 0.8989 | 0.991 |
Oligo32-4 | 0.0773 | 0.8182 | 0.988 | 0.0786 | 0.5428 | 0.960 |
Oligonucleotide | TEA/PA | TEA | Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|---|---|---|
K4 | K5 | K4 | K5 | K4 | K5 | K4 | K5 | ||
A10 | 0.010 | 9.881β | 0.0323 | 7.468β | C8 | 0.017 | 1.967β | 0.0640 | 1.639β |
A20 | 0.013 | 13.333β | 0.0401 | 11.025β | C9 | 0.021 | 2.380β | 0.0782 | 2.082β |
A25 | 0.016 | 13.568β | 0.0439 | 11.990β | C10 | 0.032 | 2.767β | 0.1049 | 2.619β |
A30 | 0.018 | 13.774β | 0.0466 | 12.755β | Oligo20-1 | 0.043 | 11.099β | 0.0607 | 12.121β |
T5 | 0.126 | 9.662β | 0.1859 | 9.766β | Oligo20-2 | 0.039 | 11.351β | 0.0632 | 12.180β |
T7 | 0.102 | 12.642β | 0.1695 | 12.270β | Oligo20-3 | 0.037 | 9.268β | 0.0386 | 11.351β |
T10 | 0.122 | 14.771β | 0.1669 | 14.577β | Oligo20-4 | 0.036 | 8.143β | 0.0366 | 9.268β |
T12 | 0.131 | 15.723β | 0.1653 | 15.625β | Oligo32-1 | 0.061 | 12.121β | 0.0928 | 12.225β |
C5 | 0.019 | 0.763β | 0.0715 | 0.679β | Oligo32-2 | 0.063 | 12.180β | 0.0922 | 12.255β |
C6 | 0.015 | 1.167β | 0.0640 | 0.934β | Oligo32-3 | 0.062 | 12.240β | 0.0904 | 12.300β |
C7 | 0.014 | 1.578β | 0.0580 | 1.271β | Oligo32-4 | 0.094 | 12.937β | 0.1448 | 12.723β |
Table 9 Quilibrium constants of K4 and K5 for oligonucleotides
Oligonucleotide | TEA/PA | TEA | Oligonucleotide | TEA/PA | TEA | ||||
---|---|---|---|---|---|---|---|---|---|
K4 | K5 | K4 | K5 | K4 | K5 | K4 | K5 | ||
A10 | 0.010 | 9.881β | 0.0323 | 7.468β | C8 | 0.017 | 1.967β | 0.0640 | 1.639β |
A20 | 0.013 | 13.333β | 0.0401 | 11.025β | C9 | 0.021 | 2.380β | 0.0782 | 2.082β |
A25 | 0.016 | 13.568β | 0.0439 | 11.990β | C10 | 0.032 | 2.767β | 0.1049 | 2.619β |
A30 | 0.018 | 13.774β | 0.0466 | 12.755β | Oligo20-1 | 0.043 | 11.099β | 0.0607 | 12.121β |
T5 | 0.126 | 9.662β | 0.1859 | 9.766β | Oligo20-2 | 0.039 | 11.351β | 0.0632 | 12.180β |
T7 | 0.102 | 12.642β | 0.1695 | 12.270β | Oligo20-3 | 0.037 | 9.268β | 0.0386 | 11.351β |
T10 | 0.122 | 14.771β | 0.1669 | 14.577β | Oligo20-4 | 0.036 | 8.143β | 0.0366 | 9.268β |
T12 | 0.131 | 15.723β | 0.1653 | 15.625β | Oligo32-1 | 0.061 | 12.121β | 0.0928 | 12.225β |
C5 | 0.019 | 0.763β | 0.0715 | 0.679β | Oligo32-2 | 0.063 | 12.180β | 0.0922 | 12.255β |
C6 | 0.015 | 1.167β | 0.0640 | 0.934β | Oligo32-3 | 0.062 | 12.240β | 0.0904 | 12.300β |
C7 | 0.014 | 1.578β | 0.0580 | 1.271β | Oligo32-4 | 0.094 | 12.937β | 0.1448 | 12.723β |
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