대사 흐름 동위원소 추적

기술 사양 & 시료

TECHNICAL SPECIFICATIONS

Item	Details
Platform	Q Exactive high-resolution MS with polarity switching and isotopomer-resolved acquisition; complementary GC-MS / LC-MS dual platforms
Samples & Systems	In vitro cell models (tumor, primary, stem cells) · in vivo animal models · microbial fermentation systems · plant tissues · plasma and other biofluids; after labeling, snap-freeze samples in liquid nitrogen and ship on dry ice
Deliverables & Timeline	Statistics tables · MID tables · analysis report; standard turnaround 4–6 weeks
Information to Confirm	Research background and questions · sample types and comparison design · target pathways · expected sample size and labeled substrates

분석 프로세스

SERVICE WORKFLOW

표지 배양

정상상태에서 동위원소 배지로 교환

MID 질량분석 검출

GC-MS / LC-MS 이중 플랫폼

천연 존재도 보정

천연 동위원소 배경 제거

네트워크 모델링

플럭스 추정 (해당 시)

통계 & 시각화

플럭스 맵, 히트맵 & 해석

실험 설계 지원:¹³C / ¹⁵N 추적자 선택과 조달, 표지 용량·기간, 시료 채취 시점 설계를 전면 지원하여 추적 비용을 통제하면서 플럭스를 측정 가능하게 유지합니다.

추적 가능 경로 & 대표 추적자

TRACER PATHWAYS

Pathway	Common Tracers / Representative Substrates · Resolvable Readouts
1 · Glucose Flux GLUCOSE FLUX	[U-¹³C]glucose / [1,2-¹³C]glucose / [1,6-¹³C]glucose / [2,3-¹³C]glucose — flux partitioning among glycolysis, the TCA cycle and the pentose phosphate pathway; [1,6-¹³C] and [2,3-¹³C] distinguish PPP branches and PDH activity; Warburg-effect quantification in tumors and proliferating cells
2 · Fatty Acid Flux FATTY ACID FLUX	[U-¹³C]acetate / [U-¹³C]palmitate — relative contributions of de novo lipogenesis (DNL) versus exogenous uptake; acetyl-CoA pools and fatty-acid β-oxidation; chain elongation and desaturation fates
3 · Nucleotide Flux NUCLEOTIDE FLUX	[U-¹³C]nucleosides/nucleotides / [¹⁵N4]hypoxanthine — balance between de novo and salvage synthesis of purines/pyrimidines; resolving the sources of proliferation-driven nucleotide demand
4 · Amino Acid Flux AMINO ACID FLUX	[U-¹³C]glutamine / [amide-¹⁵N]glutamine / [U-¹³C]serine — nitrogen flow through glutaminolysis, TCA anaplerosis and transamination; carbon-skeleton exchange among amino acids
5 · Folate / One-Carbon Flux ONE-CARBON FLUX	[U-¹³C]serine / [U-¹³C]glycine — carbon-source assignment for one-carbon units, purine synthesis and methylation; folate-cycle efficiency assessment
6 · Urea Cycle Flux UREA CYCLE FLUX	¹⁵N-NH₄Cl / [amide-¹⁵N]glutamine — nitrogen metabolism and urea-cycle flux; ammonia-clearance efficiency and ¹⁵N fate in purine synthesis

생물정보 분석

BIOINFORMATIC MODULES

Module	Methods & Metrics · Key Outputs
Data Preprocessing	Metabolite peak extraction and integration, mass-isotopomer (M+0 / M+1 / M+2 …) signal extraction, sample naming and group mapping, low-quality peak filtering — raw and corrected MID data tables
Natural-Abundance Correction	Matrix-based subtraction of natural ¹³C / ¹⁵N / ¹⁸O / ²H backgrounds; correction quality assessed by residual RMS — RMS bar charts, before/after comparison plots
Labeling Overview	Labeling rate (enrichment) = weighted mean labeling across m+ isotopologues; group-mean comparison and inter-sample correlation matrix — labeling-rate heatmap, group-mean bar charts
MID Composition	Inter-sample isotope-ratio distributions, intra-sample m+ distributions, group MID composition comparison (mean ± SD) — distribution plot sets, MID composition tables
Group Comparisons & Cross-Comparison Summary	Independent statistics per comparison (labeling-rate differences, differential MIDs); cross-comparison summary heatmaps and Venn diagrams — differential labeled-metabolite lists and intersection lists

결과 예시

SELECTED FIGURES FROM ACTUAL REPORTS

보정 잔차(RMS) 막대 차트

대사물질별 천연 존재도 보정 잔차로, RMS가 작을수록 보정 신뢰도가 높습니다.

그룹 평균 표지율 막대 차트

그룹 간 평균 표지율(농축도)을 비교하여 그룹 간 플럭스 차이를 보여줍니다.

차이 MID 히트맵

차이 대사물질의 m+ 비율을 군집화한 히트맵으로, 빨간색과 파란색이 표지 차이의 방향과 크기를 나타냅니다.

표지율 히트맵

대사물질 × 시료 표지율 행렬로, 표지가 높은 경로와 이상 시료를 빠르게 찾을 수 있습니다.

KEGG 경로 비율 분석

검출된 표지 대사물질의 KEGG level-3 경로 비율로 플럭스 커버리지 범위를 명확히 보여줍니다.

시료 내 동위원소 분포

단일 시료 내 각 대사물질의 m+ 구성으로, 결과 판독의 기본 단위입니다.

시료 간 동위원소 비율 분포

시료 간 m+ 비율 분포로, 표지가 정상상태에 도달했는지 평가합니다.

그룹 MID 구성 비교

그룹 MID 평균 ± SD를 누적 막대로 비교하여 경로를 따른 표지 전파를 비교합니다.

표지율 차이 막대 차트

지정된 비교에 대한 표지율 차이와 유의성을 제시하여 차이 표지 대사물질을 선별합니다.

적용 분야

APPLICATIONS

종양 대사 재프로그래밍해당 과정 / TCA 회로 플럭스동위원소 추적자 대사 흐름 매핑세포 에너지 대사약물 대사 기전

FAQ

How long does a Metabolic Flux & Isotope Tracing project take?

Mature targeted panels typically take 1-3 weeks; untargeted projects take 2-4 weeks (counted from sample receipt and QC approval). The exact timeline is confirmed in writing before project start, and progress is trackable on the OmicsHub cloud platform.

What if my sample amount is below the recommendation?

Please do not dilute your samples. Low-input samples can often be accommodated by adjusting the preparation workflow — tell us the actual amount and our technical team will assess feasibility and note adjustments in the study plan.

Can I add targets to an existing panel?

Yes. Mature panels support on-demand additions; novel analytes outside the list can be implemented through custom method development (requires standards and extra validation time) — see our Custom Targeted Metabolomics service.

What samples do you need for Metabolic Flux & Isotope Tracing?

Serum/plasma ≥100 μL (fasting, no haemolysis), tissue ≥50 mg (snap-frozen), cells ≥1×10⁶ (medium washed off), urine ≥200 μL. Keep fasting time consistent, aliquot into single-use volumes and avoid freeze–thaw. Full guidance: Technical Resources → Sample Guide.