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BioTechniques Volume 67, 2019 - Issue 5
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Presoaking Dried Blood Spot with Water Improves Efficiency for Small-Molecule Analysis

You-Rim Lee1 Department of Senior Healthcare,BK21 Plus Program, Graduate School,Eulji University,Daejeon34824,Republic of KoreaView further author information
,
Jiyeong LeeView further author information
&
Hee-Gyoo Kang1 Department of Senior Healthcare,BK21 Plus Program, Graduate School,Eulji University,Daejeon34824,Republic of Korea;2 Department of Biomedical Laboratory Science,College of Health Sciences,Eulji University, Seongnam-si,Gyeonggi-do13135,Republic of KoreaCorrespondence[email protected]
View further author information
Pages 219-228 | Received 06 Apr 2019, Accepted 31 May 2019, Published online: 21 Oct 2019

Figures & data

Figure 1. Image at 30 min post-incubation.

The tubes in the upper line belong to the P group (conventional method). Bloodstains were incubated for 30 min in the extraction solvent. The tubes in the lower line belong to the W group. Bloodstains were incubated for 30 min in water before adding the extraction solution. The P and W group only differ in this aspect. The sampling process (blood collection, internal standard addition and spotting) and extraction process (vortex, sonication, centrifugation, obtaining supernatant) were the same in both groups.

A: Acetonitrile; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Figure 1. Image at 30 min post-incubation.The tubes in the upper line belong to the P group (conventional method). Bloodstains were incubated for 30 min in the extraction solvent. The tubes in the lower line belong to the W group. Bloodstains were incubated for 30 min in water before adding the extraction solution. The P and W group only differ in this aspect. The sampling process (blood collection, internal standard addition and spotting) and extraction process (vortex, sonication, centrifugation, obtaining supernatant) were the same in both groups.A: Acetonitrile; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.
Figure 2. Venn diagrams of molecular features for each condition in positive ion mode.

A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Figure 2. Venn diagrams of molecular features for each condition in positive ion mode.A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.
Figure 3. Venn diagrams of molecular features for each condition in negative ion mode.

A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Figure 3. Venn diagrams of molecular features for each condition in negative ion mode.A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.
Figure 4. Dendrograms of samples for each condition.

(A–E) Results of positive ion mode. (F–J) Results of negative ion mode.

A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Figure 4. Dendrograms of samples for each condition. (A–E) Results of positive ion mode. (F–J) Results of negative ion mode.A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.
Figure 5. K-means clustering results for each condition.

The K value was set to 2. (A–E) Results of positive ion mode. (F –J) Results of negative ion mode.

A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Figure 5. K-means clustering results for each condition.The K value was set to 2. (A–E) Results of positive ion mode. (F –J) Results of negative ion mode.A: Acetonitrile; B: Blood group; M: Methanol; MA: M and A (1:1, v/v); MWF: M, water, and formic acid (80:20:0.1, v/v); MW: M and water (8:2, v/v); P: Conventional method group; W: Modified method group.

Table 1. Relative recovery for common MFs of conditions in positive ion mode.

Table 2. Relative recovery for common molecular features of conditions in negative ion mode.

Table 3. Absolute recovery and extraction efficiency of internal standards.

Supplemental material

Authors’ original file for figure 1

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