Chloroform

For cultured cell samples, cells were collected and lysed in 300 μl of TRIzol reagent (Invitrogen, U.S.A.). The samples were allowed to stand for 5 min at room temperature, after which 180 μl of Chloroform (Kanto Chemical, Japan) was added to allow for phase separation by centrifugation at 12,000 × g for 15 min at 4 °C. Next, aqueous phase was transferred to a fresh tube with equal volumes of isopropanol and GlycoBlue Coprecipitant (Invitrogen, U.S.A.). The samples were incubated at room temperature for 20 min. The samples were pelleted through centrifugation at 12,000 × g for 15 min at 4 °C. The RNA pellet was washed with 100% ethanol, air-dried before reconstituting it in nuclease-free water (Ambion, U.S.A.).

Silver acetate (CH₃COOAg), silver nitrate (AgNO₃), sodium borohydride (NaBH₄), and chloroform were obtained from Kanto Chemical Co., Inc. (Tokyo, Japan) Gold (III) chloride trihydrate (HAuCl₄·3H₂O), tetraoctylammonium bromide (TOAB), and dodecanethiol (DDT) were purchased from Nacalai Tesque Inc. (Kyoto, Japan), Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), and Sigma-Aldrich (St. Louis, MO, USA), respectively. All chemicals were used as received, without further purification.

The following reagents were used as received: N,N-dimethyl formamide (DMF), chloroform (CHCl₃), ethyl acetate (EtOAc), n-hexane, sodium sulfate (Na₂SO₄), methanol (MeOH), methylene chloride (CH₂Cl₂), sodium bicarbonate (NaHCO₃), sodium hydroxide (NaOH), N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl) uronium hexafluorophosphate (Kanto Chemical Co., Inc., Tokyo, Japan), N-ethyl-N-(1-methylethyl)propan-2-amine (Wako Co., Tokyo, Japan), stearylamine, silver (I) carbonate, N,N′-dicyclohexyl carbodiimide (DCC), 1-hydroxybenzotriazole (HOBt) (Kishida Chemical Co., Oshaka, Japan), di-tert-butyl-dicarbonate (TCI Co., Tokyo, Japan), hydrobromic acid (acetic acid solution) (Merck KGaA Co., Tokyo, Japan), α-d-cellobiose octaacetate, α-d-Glucose pentaacetate, ethyl 6-hydroxy hexanoate, dimethyl sulfoxide (DMSO), and deuterated solvents (CDCl3, DMSO-d₆ and D₂O) (Sigma-Aldrich, St. Louis, MO, USA). N,N-diisopropyl ethylamine (DIEA) were purchased from Wako Pure Chemical Industry Co., Inc. (Tokyo, Japan). Di-Boc-lysine (2) was synthesized according to the previous literature [24 (link)].

DOPC, DPPC and Chol were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Rhodamine B 1,2-dihexadeconyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (Rho-DPHE) (λ_ex = 560 nm, λ_em = 580 nm) and 6-dodecanoyl-2-dimethylamino naphthalene (laurdan, λ_ex = 430–455 nm, λ_em = 490–540 nm) were from Invitrogen (Carlsbad, CA, USA) and Funakoshi Co., Ltd. (Tokyo, Japan), respectively. Capsaicin was obtained from Sigma Aldrich Corporation (Tokyo, Japan). Chloroform and methanol were purchased from Kanto-Chemical Co., Inc. (Tokyo, Japan) and Nacalai Tesque, Inc. (Kyoto, Japan), respectively. All chemicals were of analytical grade and used as received. Deionized water was obtained using a Millipore Milli-Q (Millipore S.A.S., Molsheim, France), purification system, conductivity 18.2 MΩ cm⁻¹.

Cordycepin, proline, Folin-Ciocalteu’s phenol, ninhydrin, glacial acetic acid, thiobarbituric acid (TBA), tricholoroacetic acid (TCA), toluene and sulfosalicylic acid were bought from Sigma-Aldrich Japan K.K., Tokyo, Japan. Ethyl acetate, methanol, and methanol plus were provided from Junsei Chemical Co., Ltd., Tokyo, Japan. Hexane, chloroform, acetone, sodium carbonate and aluminum (III) chloride hexahydrate were obtained by Kanto Chemical Co. Inc., Tokyo, Japan.

Acetonitrile, chloroform, isopropanol, and methanol were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan) for the LC/MS analyses. Formic acid, ammonium formate (1 mol/L) and phenyl isothiocyanate (PITC) were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Purified water was obtained from a Milli-Q Gradient system (Millipore, Billerica, MA, USA). Ethanol was purchased from Nacalai Tesque (Kyoto, Japan). The mixture of ISs (UltimateSPLASH^TM ONE) and the chemical standards of lipid species were obtained from Avanti Polar Lipids Inc. (Alabaster, AL, USA) and Millipore-Sigma Ltd. (Burlington, MA, USA). Pooled normal human plasma, Na EDTA (IPLA-N, Lot: 26393), was purchased from Innovative Research, Inc. (Novi, MI, USA). The reference material (NIST SRM 1950 plasma, NIST QC) [61 (link)] was purchased from Merck Sigma-Aldrich (Darmstadt, Germany).