US 10012574

I'll search for authoritative records on this patent number, including USPTO and any 2026 CAFC docket activity.

Let me check for any litigation or CAFC docket activity involving this patent.

Let me look for the specific claim set and any related-family activity.

US Patent 10,012,574 — Analyst Summary

Sourcing note: The authoritative full text you supplied (Google Patents, fetched 2026‑09‑24) is the primary source for everything below. My live searches corroborated the bibliographic data (the patent appears in family/citation listings at Google Patents, FreePatentsOnline, and a Chinese patent database) but returned no litigation, PTAB, or Federal Circuit docket entries tied to this patent number. I flag uncertainties explicitly.

Bibliographic data

Abstract (as issued)

"Provided herein is a method comprising one or more of the following steps: (a) lysing cells of a biological sample and contacting the biological sample with an amount of ionic liquid sufficient to denature intracellular metabolic enzymes in the biological sample to produce a contacted cellular sample; (b) mixing the contacted cellular sample with an organic solvent to produce an ionic liquid‑organic solvent composition; (c) mixing the contacted cellular sample with the organic solvent to produce a dispersed microdroplet ionic liquid‑organic solvent composition; (d) contacting the ionic liquid‑organic solvent composition with an ion exchange composition to produce a second ionic liquid‑organic solvent composition; (d) separating the ionic liquid from the organic solvent; and (e) extracting metabolites from the ionic liquid. Kits and systems for practicing the subject methods are also provided."

(Note: the duplicate "(d)" labeling is in the original abstract text as published — I have not corrected it.)

Plain‑language overview of the disclosure

The invention is a metabolomics sample‑preparation workflow. The core insight is that an ionic liquid (IL) can do two jobs at once: (1) rupture cells, and (2) denature the cells' metabolic enzymes essentially instantaneously, thereby "quenching" metabolism so metabolite levels are frozen at the moment of sampling rather than degrading during preparation.

The workflow (spelled out in the patent's Example 1):

1. Filter a cell suspension to strip away culture media/extracellular material.
2. Resuspend the cells in a hydrophilic ionic liquid → simultaneous lysis + enzyme denaturation + metabolic quench (the patent states ≥95%–99.9% of metabolic processes can be quenched, and denaturation can be "instantaneous" when IL is added at ≥2×, ≥5×, or ≥10× by volume).
3. Add a hydrophobic/non‑polar organic solvent (e.g., pentane, hexane, heptane, octane, diethyl ether, chloroform) → two‑phase system.
4. Agitate (vortex/sonicate/shake) to form a dispersed microdroplet emulsion (0.001–1000 µm droplets) without any added dispersant, greatly increasing surface area. Hydrophobic species (lipids, membrane components) partition into the organic phase; proteins precipitate; hydrophilic metabolites stay in the IL.
5. Add an ion‑exchange composition (the working example is lithium bis(trifluoromethanesulfonimide), LiNTf₂) to trigger a salt‑metathesis reaction. The IL cation pairs with the NTf₂⁻ anion, forming a new, denser, water‑immiscible ionic liquid that coalesces and separates cleanly from the organic layer.
6. Separate and recover metabolites — remove residual IL by solid‑phase microextraction/chromatography under conditions where the IL is not an elutropic solvent; then analyze (LC‑MS, GC‑MS, etc.).

Being a "DLLME" (dispersive liquid‑liquid microextraction) method, it is a miniaturized, low‑solvent approach. The specification also describes optional downstream processing (protein capture on amine‑reactive solid supports, RNA/glycan capture via periodate oxidation to hydrazide beads, DNA purification, size‑exclusion separation) and a "three‑phase" variant in Examples 2 and 3.

Independent claims — plain‑language overview (with a hard caveat)

Uncertainty flag: The full text you supplied renders the specification, definitions, and examples, but the numbered claims section was not included in the fetched material, and my searches did not return the granted claim text. I therefore cannot quote or count the independent claims authoritatively. What follows is a plain‑language reconstruction based on the Summary, Abstract, and the system/kit descriptions — treat claim scope and count as unverified.

Based on that material, the patent appears to carry at least three independent claim families:

• Independent method claim (likely claim 1): A method of preparing a metabolomic sample in which cells of a biological sample are lysed and contacted with an amount of ionic liquid sufficient to denature the intracellular metabolic enzymes, producing a "contacted cellular sample." In plain terms: use an ionic liquid as a combined lysis/quench reagent.
• Independent method claim covering the fuller workflow: The stepwise method (a) lysing + IL contacting → (b) mixing with organic solvent → (c) agitating into a dispersed microdroplet IL‑organic composition → (d) contacting with an ion‑exchange composition to produce a second IL‑organic composition → (e) separating IL from organic solvent → (f) extracting metabolites from the IL. In plain terms: IL quench, then DLLME cleanup, then metathesis to break the emulsion, then recover metabolites.
• Independent system claim: A "system for high throughput analysis of cellular metabolites" comprising a contacting apparatus (e.g., Eppendorf tube, test tube, glass flask, 96‑well plate) for contacting samples with ionic liquid, a sampling device feeding samples to it, and an ionic‑liquid solvent chamber feeding IL to it — optionally with organic‑solvent chamber, agitator, ion‑exchange chamber, and a sample analyzer (LC‑MS/GC‑MS), plus a computer with modules (algorithms) that automate those steps.
• Independent kit claim: A kit containing one or more ionic liquids, one or more organic solvents, and an ion‑exchange composition, typically with instructions and containers/vials/pipets/filters.

The ionic‑liquid genus disclosed centers on a 1,3‑disubstituted imidazolium cation of Formula (I), with the specific working example 1‑butyl‑3‑methyl‑imidazol‑3‑ium (BMIM); the specification also names sulfonium, phosphonium, tetraalkylammonium, and pyrazolium cations and specifically lists 1,2,4‑trimethylpyrazolium methylsulfate, methyl‑trioctylammonium bis(trifluoromethylsulfonyl)imide, trihexyltetradecylphosphonium bromide, and 5‑(trifluoromethyl)dibenzothiophenium trifluoromethanesulfonate.

Litigation / CAFC 2026 docket status

I searched for Federal Circuit and district‑court activity tied to 10012574 and found nothing specific to this patent. Important negative finding: the well‑publicized 2026 Agilent patent case — Agilent Technologies, Inc. v. Axion BioSystems, Inc., D. Del. C.A. No. 23‑198 (trial scheduled March 23, 2026; pretrial order filed March 2026) — asserts U.S. 7,192,752, 7,468,255, and 8,026,080 (plus Lanham Act and DTSA claims), not the '574 patent. An older Agilent v. J&X Technologies, N.D. Tex. No. 3:19‑cv‑02156, also does not appear to involve this patent, though I could not fully verify the patents‑in‑suit there before reaching my search limit.

Bottom line: Based on available sources, US 10,012,574 appears to be an unasserted, active Agilent patent with no 2026 CAFC docket activity.

Related Agilent filings (context, not claim scope)

• US 2014/0273080 A1 — the pre‑grant publication of this application.
• US 2019/0293530 A1 ("Mass spectrometry compatible salt formation for ionic liquid sample preparation") — a later Agilent application that expressly incorporates US 2014/0273080 and US 2015/0369711 by reference and builds on the same IL‑metathesis approach, but claims priority only to 2018 provisionals. I have not verified whether US 2015/0369711 is a continuation in this '574 family.

I'll search for litigation involving US patent 10012574 across patent litigation sources.

The general searches returned no relevant results. Let me try patent litigation-specific sources.

No litigation results yet. Let me try additional litigation-specific sources.

Summary of Findings — Litigation Involving U.S. Patent No. 10,012,574

No litigation was found.

Based on my searches of patent-litigation-oriented sources (including Unified Patents' litigation/PTAB portal, Justia/CAFC case pages, Docket Alarm/PTAB materials, and general litigation-indexed web content), I could not identify any district-court lawsuit, ITC action, CAFC appeal, or PTAB proceeding (IPR/PGR) involving U.S. Patent No. 10,012,574. Accordingly, there are no cases for which I can report a plaintiff, defendant, jurisdiction, case number, filing date, or outcome.

Important caveats on what the searches did and did not show

1. The searches executed were not the litigation databases themselves. I was able to reach the Unified Patents-related litigation/PTAB materials and some CAFC decisions via search, but I could not directly run a query against the Unified Patents portal case list, PACER, or Docket Navigator for this specific number before my search budget was exhausted. So my statement is "no litigation identified," not a verified "no litigation exists." A definitive negative would require a direct docket query (PACER/Docket Navigator/Lex Machina) on the exact number.

2. Hits containing "10012574" as a string were false positives and were excluded. The literal string "10012574" appeared in results that have nothing to do with this patent — e.g., a Brazilian concurso candidate-registration PDF and a Cleveland Metropolitan School District financial report ("CON-10012574: Artist-In-Residence"). Per your instruction to interpret identifiers literally and not auto-correct, I did not treat these as the patent number, and I did not substitute a "similar" number.

3. No similar-number results were returned. My searches for "<10,012,574>"-type variants and for Agilent enforcement actions returned none, so there was nothing to confuse with the target patent.

Patent identity confirmation (for docket-query accuracy)

The number you gave resolves, per the authoritative full text supplied, to:

• U.S. Patent No. 10,012,574 B2
• Title: Method for metabolomic sample preparation based on ionic liquid dispersive liquid-liquid microextraction
• Application No.: US 14/205,100
• Inventor: James Alexander Apffel, Jr.
• Original/current assignee: Agilent Technologies, Inc.
• Priority: 2013-03-15; Filed: 2014-03-11; Granted: 2018-07-03
• Pre-grant publication: US 2014/0273080 A1
• Status: Active; adjusted expiration 2037-01-07

Note that assignee/status information on the patent record is expressly stated there as an assumption and not a legal conclusion, so it should not be relied on for litigation purposes without independent verification.

Recommended next steps to confirm a true negative

To definitively confirm whether this patent has ever been asserted or challenged, query by the exact identifiers above in:

• PACER (civil docket search on "10,012,574" and on assignee "Agilent Technologies") and CourtListener/RECAP;
• Unified Patents Portal — both the litigation case list (portal.unifiedpatents.com/litigation/caselist) and the PTAB case list (portal.unifiedpatents.com/ptab/caselist);
• USPTO Patent Center / PTAB for any IPR, PGR, or reexamination tied to application 14/205,100;
• Commercial docket tools (Docket Navigator, Lex Machina) filtering on the patent number.

If you'd like, I can attempt targeted queries against each of those specific resources (one at a time) to firm up the negative or surface any case I may have missed.

Proceedings overview

Zero AIA trial proceedings. The structured "PTAB proceedings on file" block — the canonical list from the USPTO Open Data Portal — returns no IPR, PGR, or CBM for U.S. Patent No. 10,012,574 (breakdown: 0 active / 0 claims invalidated / 0 claims sustained / 0 settled / 0 institution denied). My independent web searches corroborate this: every string hit for "10012574" resolved to unrelated documents (a Brazilian concurso PDF, a Cleveland schools financial report, and unrelated PTAB/§ 325(d) papers), and no Agilent or third-party PTAB paper references this patent. Defensive posture: the claims are untested at the Board — no cancellation, no estoppel, and no helpful FWD to lean on. You will have to build any invalidity case from scratch on the specification and prosecution history.

(Cross-reference to the previously generated sections: the "no litigation" finding there is consistent with this result. An unasserted patent is exactly the profile of a patent that attracts no IPRs — so this absence is a coherent signal, not a data gap.)

Proceedings

None. There is nothing to report at proceeding granularity — no proceeding number exists to cite, and I will not invent one. For completeness, the AIA-trial posture of this patent by vehicle:

Strategic summary

Claim status: 100% UNTESTED. No claim of US 10,012,574 has been canceled, confirmed, or construed by the PTAB. Nothing in this patent's file has been narrowed by an IPR certificate, so the claim language as issued (as reconstructed from the specification in the earlier sections — the numbered claims were not in the fetched text, so I still cannot quote or count them authoritatively) is what you face. Do not tell a client that any claim of this patent is dead, or that this patent is "hardened." Neither statement is supported. The correct characterization is "challenged by nobody, so far."

Estoppel landscape: none. With zero proceedings, § 315(e)(2) estoppel has never attached to anyone. No petitioner, no privy, and no real party in interest is barred from raising any § 102/§ 103 ground. This cuts both ways for you: (a) you are free to file an IPR on any patent-and-printed-publication ground, and (b) if you do file one and lose, you personally take the estoppel hit for grounds you raised or reasonably could have raised. Because the art is likely to include non-patent literature (the ionic-liquid DLLME and IL-denaturation literature is substantial and largely academic), consider scouting the strongest printed-publication grounds before committing to an IPR, since anything you can reasonably raise but leave out of the petition is compromised at the district court.

Pattern signals: not applicable across this patent, but note the patent owner's profile. No petitioner has filed serial IPRs against '574, and there is no defensive aggregator (e.g., Unified Patents) in the chain. However, the searches surfaced two Agilent post-grant patterns worth carrying into any engagement: (1) Agilent as habitual petitioner — IPR2018-00297, -00298, -00299 and -00313 against Thermo Fisher Scientific over RE45,553, RE45,386, and 7,230,232 (see the parallel denial arguments in the Thermo IPR2018-00313 papers and the institution decision in IPR2018-00299); and (2) Agilent as patent owner losing at the Board — in Synthego Corp. v. Agilent Techs., IPR2022-00402 and IPR2022-00403, the Board issued final written decisions on 2023-05-17 holding all claims of both asserted patents unpatentable, and Agilent appealed (oral argument 2025-03-07; the related N.D. Cal. case 5:21-cv-07801-EJD remained stayed as of the 2025-05-01 joint status report). Those are different patents and supply no claim-level precedent for '574 — I list them only to show that (i) this assignee both uses and is targeted by the AIA trial system, and (ii) the '574 portfolio has not yet been drawn into that pattern.

Portfolio note (from the earlier sections, flagged as relevant to freedom-to-operate, not to PTAB): the later Agilent application US 2019/0293530 A1 ("Mass spectrometry compatible salt formation for ionic liquid sample preparation") expressly incorporates US 2014/0273080 (the pre-grant publication of this '574 application) by reference and continues the same IL-metathesis workflow. If you are mapping the family, treat '574 as an early-generation node in a still-active Agilent IL-sample-prep line — but note that the incorporation by reference is a specification-support fact, not a PTAB fact, and I have not verified whether US 2015/0369711 is a continuation in the '574 family.

Recommended next steps

1. Confirm the negative at the source of record. Query the patent-specific PTAB E2E ("PATTS") page and the PTAB case list for application 14/205,100 and patent 10,012,574: PTAB E2E / PATTS and Unified Patents PTAB case list. The ODP ingest behind the structured block can lag newly filed petitions; if you find a recently filed (not yet institution‑decided) petition, that changes the posture materially and should be treated as the top of the stack.
2. Because there is no FWD to quote, build your own claim chart from the issued claims. The fetched full text did not include the numbered claims, so the highest-value next action is pulling the claim set from USPTO Patent Center (application 14/205,100) or the granted PDF, and reading the claims against the specification's Example 1 (filter → resuspend in hydrophilic IL → add hydrophobic organic solvent → agitate to microdroplets without added dispersant → add LiNTf₂ metathesis → separate → solid-phase microextraction). The "no added dispersant" and "second ionic liquid-organic solvent composition" limitations are the natural places to look for § 112(b) indefiniteness and written-description gaps — but those are district-court-only fights now, because the PGR window closed on 2019-04-03.
3. Mind the § 315(b) clock. This patent has no litigation history that I could find, so no one is currently time-barred. The moment a complaint asserting '574 is served on you, the one-year IPR window opens; if the asserted claims are broad method claims with thin written-description support, an early petition may be worth reserving.
4. Do not overstate the negative. "No PTAB activity" is a verified absence as of the ODP ingest date and my searches, not a guarantee. Say it plainly and give the caveat, rather than analogizing to patents that have been tested.

I'll search for the assignment record for this patent number.

Let me try to reach the assignment record directly and via other indexed sources.

Let me try to pin down the recorded assignment's reel/frame and correspondent.

Assignment & Ownership Reconstruction — US 10,012,574

Sourcing note and hard caveat up front. I was able to retrieve the patent's legal-events record (via Google Patents, which mirrors USPTO assignment data) and corroborating pages (Justia, patentimages PDF). I was not able to directly query https://assignmentcenter.uspto.gov/ or https://assignment.uspto.gov/patent/index.html in this session, so I do not have the Reel/Frame number, the execution date, or the correspondent of record for the one assignment that exists. I am not going to invent those fields. Where a field is unavailable I mark it [not retrieved]. Everything else below is grounded in the record and cross-referenced to the litigation/PTAB findings in the earlier sections of this analysis.

This patent has a recorded assignment chain — it is not a "no records" case. There is exactly one recorded conveyance, the original inventor-to-employer assignment. That is the entire post-filing ownership history.

Inventors

Pattern check — departures: No unusual pattern. A single inventor is named, so the "all inventors departed within 12 months" tell cannot even be evaluated as a group. More importantly, the record leans against a departure: Apffel continues to appear as inventor on later Agilent-assigned applications in the same technical space — e.g., an application directed to "A method for removing an ionic liquid from an aqueous sample" (US 2015/0369711 family), also assigned to Agilent Technologies, Inc. (Justia inventor page: https://patents.justia.com/inventor/james-alexander-apffel). Continued filing activity assigned to the same employer is inconsistent with a fire-sale-antecedent departure. [Verdict: not present.]

Minor cross-reference note: the earlier "Patent summary" section flagged that US 2015/0369711 may or may not be a continuation of this '574 family. The inventor overlap and shared assignee support a same-family/same-inventor-group relationship, but I still cannot confirm a formal continuation priority link from the material I have — treat that as unresolved.

Original assignee

Agilent Technologies, Inc. (Delaware corporation) — named as original and current assignee on the issued patent. Per the patent record supplied, assignee status is expressly stated to be an assumption and not a legal conclusion.

• Primary line of business: analytical instrumentation and life-sciences/chemical-analysis products — LC‑MS/GC‑MS systems, chromatography columns, and consumables. This is directly aligned with the patent's subject matter (a mass-spectrometry metabolomics sample-prep method).
• Status: Operating, publicly traded, solvent. Agilent (NYSE: A) is the 1999 HP spin-off; it remains an independent operating company. There is no indication of acquisition, dissolution, bankruptcy, or name change in the record.
• Did they ship a product embodying the claims? Unclear / not established. The claims are method claims (a metabolomics sample-prep workflow). Agilent indisputably sells the instrument ecosystem the method feeds (LC‑MS/GC‑MS) and sells sample-prep consumables, but I have no verified evidence of a specific Agilent commercial kit or product that practices the full claimed IL‑DLLME workflow. I will not assert product embodiment I can't document.
• SEC cross-reference: Agilent's 10‑K/8‑K filings do not itemize individual patents, and I found no SEC disclosure referencing this patent or any patent-sale collateral arrangement involving it. So SEC filings neither confirm nor contradict the assignment record — they are simply silent at this granularity.

Assignment timeline

Chronological list of every recorded assignment:

• executed [not retrieved] / recorded 2014-03-17 — Reel [not retrieved]/[not retrieved]
  • Conveyance: Assignment of assignors' interest ("ASSIGNMENT OF ASSIGNORS INTEREST")
  • Assignor: James Alexander Apffel, Jr.
  • Assignee: AGILENT TECHNOLOGIES, INC.
  • Correspondent: [not retrieved] — I could not access the correspondent-of-record field. It cannot be flagged as a repeat player because I have no name to compare.
  • Context: Routine inventor-to-employer assignment, recorded six days after the 2014-03-11 application filing. This is the standard "assignment on filing" pattern for a large corporate filer, not a transfer between unrelated parties.

That is the full chain. No subsequent assignment (change of name, merger, security agreement, license, release, or correction) appears in the legal-events record. There is no conveyance to any IP-holding LLC, no security interest recorded against the patent, and no release or reversion. The Google Patents legal-events block shows the 2014-03-17 assignment as the sole reassignment event through issuance (2018-07-03) and as of the fetch date (2026-09-24).

Verification links:

• Assignment Center (patent-number search): https://assignmentcenter.uspto.gov/ — enter 10,012,574
• USPTO legacy assignment index: https://assignment.uspto.gov/patent/index.html — search 14/205,100 or 10012574
• Google Patents legal events (source actually used): https://patents.google.com/patent/US10012574/en

Timeline diagram

timeline
    title Ownership of US 10012574
    2013 : Priority provisional filed by Apffel
    2014 : Non-provisional filed by Apffel
         : Assigned to Agilent Technologies
    2018 : Patent issued to Agilent
    2037 : Adjusted expiration

NPE / troll-pattern signals

Verdict

Insufficient data (only the original assignment).

Per the required taxonomy, "only the original assignment" is the defined trigger for this verdict, and that is exactly the record here: a single conveyance, James Alexander Apffel, Jr. → Agilent Technologies, Inc., recorded 2014-03-17, with no reel/frame or correspondent retrievable in this session. I want to be precise about why this is the right call rather than "NPE" or "operating-company assertion": no NPE signal is present (0 of 8), so an NPE verdict would be affirmatively wrong; but the "operating-company assertion" verdict requires the owner to be suing actual competitors on this patent, and the earlier litigation section established no suit naming US 10,012,574 exists. With one recorded link, no plaintiff, and no post-issuance transfers, the chain simply provides nothing to score as a troll pattern.

Bottom line for an ownership analyst: this is a clean, unremarkable employee-assignment chain that terminates at the original operating-company assignee, Agilent Technologies, Inc. There is no shell entity, no assertion vehicle, no bankruptcy or fire-sale history, and no defensive-aggregator neutralization. The only "finding" is an evidentiary gap — the reel/frame and correspondent fields — which should be closed by a direct Assignment Center query on 10,012,574 / 14/205,100 before this ownership record is relied on for any standing or chain-of-title purpose.

I'll research the prior-art citations for US 10,012,574. Let me run several targeted searches.

The general searches returned unrelated material. Let me target the patent's own citation lists directly.

The citation lists aren't appearing in general results. Let me try to pull the patent's own citation tables and the examiner-cited references.

Prior-Art Analysis — US 10,012,574 B2

Bottom line up front: I was unable to retrieve the actual prior-art citation list (the front-page "References Cited" / "Citations" tables) for US 10,012,574 from the sources available to me, and the authoritative full text supplied earlier does not include that section (it renders the abstract, definitions, description, and examples but stops before the claims and does not carry the front-page reference tables). Because your instructions require that I not fabricate, I am not going to invent a list of references or assign § 102 claim mappings to references I have not verified. Below I separate (A) what is verifiable from the material I have, (B) what I found that is not prior art, and (C) genuinely relevant field art that I can flag as a candidate only.

Contradiction flag: This task presupposes a retrievable set of "patent citations for 10012574." That premise conflicts with the record I actually have — the patent text supplied lacks both the front-page reference list and the numbered claims. I am reporting the gap rather than filling it with guesses.

A. What IS verifiable from the authoritative text

A.1 Patent identity (matches the earlier bibliographic section — no contradiction)

A.2 References cited within the specification (NOT asserted prior art)

The specification cites six U.S. patents — but expressly as background on mass-spectrometer hardware, in the boilerplate sentence: "Such systems are well known in the art (see, e.g., U.S. Pat. Nos. ...)". These are technique/context citations, not novelty-defeating art, and I would not map them to any claim under § 102:

• U.S. 7,534,996
• U.S. 7,531,793
• U.S. 7,507,953
• U.S. 7,145,133
• U.S. 7,229,834
• U.S. 6,924,478

I have not verified the dates/assignees of these six from a primary source in this session, so I am giving the numbers exactly as they appear in the patent text and declining to state publication/filing dates I have not confirmed. Critically, all six predate the 2013 priority date, so they are at least eligible as § 102 references — but they describe MS instrumentation, not the claimed ionic-liquid lysis/quench/DLLME metabolomics method, so they would not anticipate the operative claims.

A.3 The specification's own admitted state of the art (non-patent literature, unnamed)

The description characterizes DLLME in general terms — "the extraction solvent is hydrophobic and of higher density than water while the disperser is miscible with both aqueous and organic phases" — with no bibliographic citations. So the patent's DLLME background is admitted prior art without an identified reference. That is analytically important: the patent concedes DLLME and its parameters were known, meaning the likely § 102/§ 103 battleground is the ionic-liquid-as-simultaneous-lysing-and-denaturing-agent feature, not the DLLME hardware.

B. Things I found that are NOT prior art (excluded)

1. "Cited By" / forward citations — The number 10012574 appears in the forward-citation tables of other documents, e.g. US 2011/0124034 A1 (Heliae Development) and US 2009/0004048 A1, where "US10012574B2 … Agilent Technologies" is listed as a later-published citation. These are documents that cite the '574 patent, not art that predates it. They are irrelevant to § 102 and I have excluded them.

2. Document JP2016521430A search report (PCT/US2013/071235) — FALSE POSITIVE. One search hit returned an international search report under PCT/US2013/071235 citing US 2009/0297404 (Shannon), JP 2002-367982 (Matsushita Electric), US 2008/0079408 (Hasegawa), US 6,509,542, and US 6,020,794. The subject matter of those citations (image-forming/electrophotographic content, figures numbered into the "eighties") is unrelated to analytical sample preparation, and PCT/US2013/071235 is a different application from the '574 family. I have not attributed these references to US 10,012,574. I flag this so no downstream reader repeats the error.

3. String-collision noise — "10012574" also appears in unrelated records (a Brazilian concurso registration list; a Cleveland Metropolitan School District financial report "CON-10012574: Artist-In-Residence"; an IEEE document URL). Per the literal-interpretation rule, these were not treated as the patent.

C. Candidate relevant prior art (SUPPLEMENTARY — unverified as to citation/§ 102)

The following are references I encountered in field literature during these searches that go to the patent's core concepts. I am not representing that any was cited during prosecution of the '574 patent or that any anticipates a specific claim — I could not verify either, and I lack the claim text to do so.

1. Rezaee, M.; Assadi, Y.; Milani Hosseini, M.R.; Aghaee, E.; Ahmadi, F.; Berijani, S. "Determination of organic compounds in water using dispersive liquid–liquid microextraction," J. Chromatogr. A 1116 (2006) 1–9. The foundational DLLME paper (appeared as ref. [108] in a 2019 review surfaced in search). It establishes DLLME with organic extractant + disperser. Potential relevance: would go to any claim reciting only generic DLLME with organic solvent/microdroplets; it does not disclose an ionic liquid, cell lysis, or enzyme denaturation, so I could not map it, even provisionally, to a claim requiring those elements.

2. Trujillo-Rodríguez, M.J.; Rocío-Bautista, P.; Pino, V.; Afonso, A.M. "Ionic liquids in dispersive liquid–liquid microextraction," TrAC Trends Anal. Chem. 51 (2013) 87–106 (appeared as ref. [5] in a search hit). Published in 2013 — date of publication relative to the 2013-03-15 priority date is a genuine open question I could not resolve, which matters for whether it is § 102(a)/(b) art at all. A review combining ILs with DLLME. Potential relevance: would bear on the IL+DLLME combination, but reviews generally do not disclose the specific lysis/denaturation "quench" step.

3. The patent text itself names specific ionic liquids (BMIM, 1,2,4-trimethylpyrazolium methylsulfate, methyl-trioctylammonium NTf₂, trihexyltetradecylphosphonium bromide, 5-(trifluoromethyl)dibenzothiophenium triflate) and the LiNTf₂ metathesis step. Any art disclosing IL-mediated protein denaturation or in-situ IL anion metathesis with LiNTf₂ to condense a dispersed IL phase would be the highest-value § 102 art — I was unable to locate or verify such references in this session, so I am not naming any.

D. Why the requested § 102 claim mapping cannot be responsibly produced here

1. No numbered claim text is available. As flagged in the earlier section, the supplied material omits the claims. Without the granted claim language I cannot say which claim (e.g., "claim 1" vs. the fuller workflow claim vs. the system/kit claims) any reference might anticipate. Mapping art to claim numbers now would be fabrication.
2. No verified citation list. I could not open the patent's own front-page "References Cited" table (Google Patents Citations section), the USPTO Patent Center prosecution record for application 14/205,100, or FreePatentsOnline/Espacenet INPADOC citing documents before exhausting my search budget. Every search for the citation list returned either unrelated families or forward citations.

Therefore, I decline to state, for any reference, that it "anticipates claim X under § 102." I have no verified reference set and no claim text to map it against.

E. How to obtain the exact citation list (so I can complete the § 102 mapping)

Run these targeted queries on the exact identifiers, then hand me the output and I will do the full per-reference § 102/§ 103 mapping:

1. USPTO Patent Public Search (https://ppubs.uspto.gov) — retrieve US 10,012,574 B2; the front page lists "References Cited" (U.S. Patent Documents / Foreign Patent Documents / Other Publications) and "Primary Examiner."
2. USPTO Patent Center (https://patentcenter.uspto.gov) — open application 14/205,100 and pull the filed/issued "List of References Cited by Examiner" (PTO-892) and any examiner's notice of references; this is the authoritative source for examiner-cited art.
3. Google Patents (https://patents.google.com/patent/US10012574B2/en) — the "Citations" tab (patent + non-patent literature cited by the examiner/applicant) and the "Similar Documents" list.
4. FreePatentsOnline (https://www.freepatentsonline.com/10012574.html) — front-page "References Cited."
5. Espacenet / INPADOC (https://worldwide.espacenet.com) — "Also published as" and cited-documents view for the family around US 2014/0273080 A1.

Please paste the reference table (or the PTO-892), and I will return: full citation, publication/filing date, a brief technical description of each reference, and — against the granted claim text, once we have it — the specific claim(s) each reference potentially anticipates under § 102, plus any § 103 combination that appears stronger.

I'll ground this analysis by pulling the patent's actual claim set and its cited prior-art references. Let me run several targeted searches in parallel.

I have the claims. Now let me get the complete claim set (claims 17+) and the patent's cited prior-art references, which is the "Prior Art section" the task points to.

I've now retrieved the claim set and the page's prior-art metadata, but I hit my search ceiling before I could pull the patent's full citation list. Here is the § 103 analysis, with sourcing limits flagged up front.

Obviousness analysis — US 10,012,574 B2 under 35 U.S.C. § 103

1. Sourcing, and a correction to the earlier sections

Two upgrades to earlier work, plus caveats — read these first.

• The claim set is now available. The previously generated "Patent summary" and "PTAB challenges" sections flagged that the numbered claims were not in the fetched text and therefore could not be quoted. I retrieved a claim set (claims 1–17) from the Patents‑Review rendering of the application at https://www.patents-review.com/a/20140273080-method-metabolomic-sample-preparation-based-ionic-liquid.html#7. Important caveat: that rendering is of the pre‑grant publication US 2014/0273080 A1, not necessarily the granted claims of US 10,012,574. The granted claims may have been amended during prosecution. I could not pull the granted claims from USPTO Patent Center before my search budget ran out. Treat the claim language below as provisional until verified against the granted PDF (http://patentimages.storage.googleapis.com/39/7f/51/2d2067ab9dbdd3/US10012574.pdf).
• The "Prior Art section of this page" is thin in what I could retrieve. The Google Patents page supplies only prior‑art keywords — substituted, ionic liquid, cells, heteroaryl, organic solvent — plus two depicted structures (a generic substituted imidazolium *N1C=C[N+]([1*])=C1, and a set including BMIM and a dicyanamide species). It also cites, for the mass‑spectrometry hardware only, U.S. Pat. Nos. 7,534,996 / 7,531,793 / 7,507,953 / 7,145,133 / 7,229,834 / 6,924,478. I could not retrieve the patent's front‑page list of cited U.S. references or non‑patent literature. That list is the single most important item to pull next, because the strongest § 103 grounds for this patent are almost certainly in the non‑patent ionic‑liquid DLLME literature, not the patent art. Everything I label [candidate art] below is drawn from well‑documented field art that I surfaced during search but could not confirm appears on the '574 face.

2. The claims at issue (from US 2014/0273080 A1 rendering)

Note the claim set does not recite the specification's "no added dispersant" limitation. That matters: the most distinctive‑sounding feature in the spec (dispersant‑free microdroplet formation) is not in the claims as rendered, so it is not a § 103 obstacle.

3. Legal standard

Under Graham v. John Deere, 383 U.S. 1 (1966), and KSR Int'l v. Teleflex, 550 U.S. 398 (2007), a claim is obvious where the differences over the prior art are such that the subject matter as a whole would have been obvious to a PHOSITA, considering (i) scope/content of the art, (ii) differences, (iii) level of ordinary skill, and (iv) secondary considerations. Critically under KSR, a motivation to combine need not be stated in the references; it may come from the nature of the problem, common sense, or the predictable use of prior‑art elements according to known methods. Where a combination is "the mere arrangement of old elements, each performing the same function it was known to perform," it is obvious. In re Aller and In re Boesch make clear that selecting an optimal or obvious‑to‑try member of a disclosed genus (e.g., picking BMIM out of an imidazolium genus, or LiNTf₂ out of a list of exchange salts) is routine optimization, not invention.

Level of ordinary skill: a trained analytical chemist/biochemist with an M.S./Ph.D. (or equivalent experience) in bioanalytical chemistry or metabolomics, familiar with (a) quenching strategies for metabolome sampling, (b) liquid‑liquid extraction and DLLME, and (c) ionic‑liquid chemistry. That artisan is the same person the patent's own Examples describe.

4. What the specification itself admits is prior art

These are applicant admissions usable as prior art regardless of whether they appear on the face of the patent:

• "Ionic liquids are suitable for denaturing proteins and have been used for the extraction of small molecules and DNA." (spec, [Background/Examples]).
• DLLME "has been widely applied to a range of analytical samples, primarily environmental," and the patent recites the classic DLLME mechanism — hydrophobic extraction solvent denser than water, disperser miscible with both phases, turbid microdroplet solution, "large contact area… remarkably decreasing the extraction time and increasing the extraction efficiency."
• Ionic liquids are "salts composed of ion pairs that are in the liquid state at room temperature" — i.e., RTILs by definition operate at room temperature (foreshadowing claim 11).
• The protein‑denaturation chemistry is framed as conventional.

So the patent concedes three of the four conceptual pillars: (1) ILs denature proteins, (2) ILs extract small molecules/DNA, (3) DLLME generates high‑surface‑area microdroplets for fast extraction. The only remaining pillar is the combination of these with ILs in a metabolomic quench workflow — and the fourth pillar, salt metathesis to reclaim the IL, is independently known in the IL‑DLLME literature.

5. The prior‑art combinations

Combination A — Claim 1 (the broadest claim; the most vulnerable)

References: An IL‑protein‑denaturation/IL‑cell‑lysis reference + a metabolomic‑quenching reference +, optionally, the patent's own admitted IL‑extraction art.

Rationale. Claim 1 requires only: lyse cells; contact with IL in an amount sufficient to denature intracellular metabolic enzymes. Nothing in the claim requires DLLME, metathesis, or any recovery step. Prior art taught that ILs (i) disrupt/denature proteins, (ii) lyse cells, and (iii) extract small molecules and DNA from biological material. A PHOSITA facing the recognized problem in metabolomics of stopping metabolism rapidly (cold‑methanol quenching, acid quenching, and filter‑freeze approaches were all known) would have had an explicit motivation to substitute an IL lysis/quench reagent: it does lysis and denaturation in one operation, and ILs were already known to be compatible with downstream small‑molecule extraction. Under KSR, using a known denaturant/lytic agent to perform its known function is not inventive. Claim 1 is the strongest candidate for a § 103 rejection — likely anticipation‑adjacent if a single reference teaches IL lysis + denaturation of a cellular sample.

Supporting field art [candidate]: the widely cited IL‑DLLME and IL‑bioanalysis literature (e.g., the review chapter at https://shodhganga.inflibnet.ac.in/bitstream/10603/472021/9/09_chapter%206.pdf, which catalogs IL‑DLLME applications to biological matrices and notes ILs' low volatility/fl ammability and extraction performance). [Recall/field knowledge, not verified against the '574 face.]

Combination B — Claims 3, 4, 5, 8 (the full workflow)

References:

1. Rezaee et al. (2006) — the original DLLME paper (dispersed microdroplet extraction; this is the foundational DLLME reference the '574 spec paraphrases almost verbatim). [Recalled; verify.]
2. An IL‑DLLME / "in‑situ IL‑DLLME" reference using a hydrophilic IL that is converted to a hydrophobic IL by anion metathesis with LiNTf₂ (the "in‑situ solvent formation" technique). [Candidate; the shodhganga review explicitly names "in‑situ IL‑DLLME… in situ solvent formation for microextraction based on ILs" and lists IL‑DLLME applications — Fan, Li, Peña, Gharehbaghi, Xu, Asensio‑Ramos, Liu, Yousefi, inter alia.]
3. A metabolomic quenching reference.

Claim mapping and motivation.

• Claim 3 (add organic solvent): Rezaee‑type DLLME teaches an aqueous sample + hydrophobic extraction solvent → two‑phase.
• Claim 4 (dispersed microdroplets): Rezaee teaches that the DLLME dispersion step is the microdroplet‑forming step, and explains the surface‑area/speed benefit — exactly the '574 rationale.
• Claim 5 (ion‑exchange → second IL‑organic composition): in‑situ IL‑DLLME teaches adding an ion‑exchange salt (classically LiNTf₂) so the hydrophilic IL exchanges anion, becomes hydrophobic/denser, and phase‑separates as a discrete IL layer — an explicit teaching of "contacting with an ion exchange composition to produce a second ionic liquid‑organic solvent composition." This is the single most probative ground against claims 5 and 7.
• Claim 8 (extracting metabolites): inherent to any DLLME/extraction step; the patent's own spec says hydrophilic metabolites "are extracted into the ionic liquid."

Motivation to combine is strong and multi‑sourced: (i) both DLLME and IL‑quenching served the same field need (fast, miniaturized, low‑solvent bioanalysis); (ii) IL‑DLLME was already applied to biological matrices (serum, urine, blood, brain tissue — see shodhganga and the J. Pharm. Biomed. Anal. / Talanta IL‑DLLME literature); (iii) the in‑situ metathesis step was a known "how to get your IL back out" solution to the known problem that ILs interfere with LC‑MS (ion suppression). A PHOSITA would have had reasonable expectation of success that combining an IL quench with an IL‑DLLME cleanup would work, because each element performs the identical function it was known to perform.

Combination C — Dependent claims 6, 7, 9, 10, 11, 12

Combination D — System claims 13–16

Claim 13 recites a contacting apparatus + sampling device + IL solvent chamber — i.e., a sample container, a sample handler, and a reagent reservoir. That is the ordinary architecture of any automated liquid‑handling/analytical instrument (the spec itself lists Eppendorf tube, test tube, flask, 96‑well plate; high‑throughput sample changer; solvent reservoir). Claims 14–16 add an ion‑exchange chamber, LiNTf₂, and an analyzer (LC‑MS/GC‑MS) — all conventional in automated sample prep. Under KSR, automating a known bench method with known robotic liquid‑handling components, each performing its ordinary function, is obvious; there is no claimed structural or functional synergy. Claim 16 in particular (a mere "sample analyzer") is a non‑limiting result‑oriented element that a PHOSITA would supply as a matter of course.

6. Motivation to combine — consolidated

A PHOSITA would combine these references because:

1. Common problem, common field. Metabolomic quenching and miniaturized extraction both target fast, reproducible, low‑volume sample prep. DLLME's whole selling point — high surface area → short extraction time (Rezaee; and the '574 spec admits this) — maps directly onto the quenching‑speed requirement.
2. ILs were already the known dual‑function reagent (lyse + denature + solvate small molecules), per applicant's own admission.
3. The metathesis step was a known solution to a known IL‑related problem — removing the IL cation to avoid LC‑MS contamination/ion suppression (the mechanism discussed at length in Agilent's own later application US 2019/0293530, §[0004]–[0006], which describes salt metathesis with a water‑immiscible IL‑forming counterion as a pre‑existing paradigm).
4. Predictable, same‑function elements. Each claim element (organic solvent, dispersion, ion exchange, LiNTf₂, room‑temperature contacting, filtering) performs the identical function it was known to perform. Under KSR that is the hallmark of obviousness.
5. No criticality. The claims recite no unexpected numerical limits, no critical ratios, no synergistic effect — the hallmarks that would rebut obviousness are absent.

7. Rebuttal / secondary considerations to check

• Unexpected results: the patent reports ≥95–99.9 % quenching and "instantaneous" denaturation. If the applicant has comparative data showing the IL method preserves metabolites that cold‑methanol quenching degrades, that is a nexus argument — but it is a results argument, and the claims recite no numbers, so the nexus to the claimed scope is weak.
• Teaching away: one could argue ILs' viscosity and LC‑MS ion suppression counsel against ILs in metabolomics — but the metathesis step (the very thing claimed) was the known fix, so this argument largely cuts toward obviousness.
• Commercial success / long‑felt need: the field did have a long‑felt need for fast quench; that supports the existence of the motivation to combine rather than rebutting it.

8. Bottom line

• Claim 1 is the lead target and the most likely to fall under § 103 (IL lysis + denaturation of intracellular metabolic enzymes is squarely within the acknowledged art).
• Claims 3–8 fall to Combination B — DLLME (Rezaee) + in‑situ IL‑DLLME metathesis (LiNTf₂) + metabolomic quenching — with the in‑situ anion‑exchange teaching being the single most probative reference against claims 5 and 7.
• Claims 6, 7, 9–12 are routine‑optimization dependents.
• System claims 13–16 are automation of a bench method using conventional components.

Do not overstate this. Two things must be verified before any of it is usable: (1) the granted claim text (the analysis above uses the published claims), and (2) the patent's actual prior‑art citation list — the strongest references for this patent are almost certainly the non‑patent IL‑DLLME / IL‑denaturation literature, which the page's keyword metadata hints at but which I could not enumerate before exhausting my search budget. I have labeled every field reference [candidate] for that reason and have not fabricated any citation number, date, or holding.