Bu araştırma temel bilim araştırma programı tarafından desteklenmiştir (proje sayısı: 2016R1A2B4013467) aracılığıyla Ulusal Araştırma Vakfı, Kore (Bilim Bakanlığı, ICT ve gelecek planlama 2016 tarafından finanse edilen ve ayrıca Kore temel bilim tarafından desteklenen NMK) Enstitüsü araştırma programı (proje sayısı: C36707).

1. DMMTAV sentezi
<ol><li>
Kimyasal hazırlama ve molar oranı karıştırma DMAV, Na2S ve H2SO4 Not: DMAV: Na2S:H2kadar4 1:1.6:1.6 =<ol>
<li>DMAV 40 mL deiyonize ve N25,24 g çözülür-tasfiye (en az 30 dk için tasfiye) 50 mL santrifüj tüpü suya.</li>
		<li>Na2S reaktif 14,41 g Na2S·9H2O 50 ml deiyonize, N2çözülerek hazırlamak-bir 250 mL şişe su tasfiye.</li>
		<li>Hazırlamak için 40 mL konsantre sülfirik asit (% 96) 3,3 mL ekleyerek4 reaktif deiyonize Yani H2ve N2-50 mL santrifüj tüpü suya temizlendi. Not: DMAVson molar oranı: Na2S:H2SO4 = 1:1.6:1.6 1,3,4,8,15.</li>
		<li>DMAV çözüm (Adım 1.1.1) hazırlanan 40 mL 50 mL 250 mL şişe (Adım 1.1.2) bulunan Na2S çözeltisi ekleyin. DMAV 10 mL tasfiye su ile içeren tüp durulayın ve bu 250 mL şişe de ekleyin.</li>
		<li>Şişeye cam tüpleri ile donatılmış bir üç delikli lastik tıpa ile kapatın. Camı kullan N2 gaz giriş, çıkış ve H2SO4 çözüm giriş, sırasıyla tüpler. Hemen şişeye kapattıktan sonra balonun içine N2 gaz akışını sağlar. Not: Gaz basıncı sıçramasına olmadan tepki çözüm yüzey üzerinde akmaya sağlanmalıdır.</li>
		<li>Aside dayanıklı H2cam tüp SO4 çözüm giriş H2hazır 40 mL kadar eklemek için 50 mL şırınga ile boru bağlanmak4 çözüm (Adım 1.1.3). H240 mL kadar ekleyin4 çözüm, yavaş yavaş ve kademeli bir şekilde. Uyarı: Sülfürik asit ekleyerek üzerine beyaz duman-ecek var olmak oluşturmak; iyi havalandırılmış duman başlığı kullan.</li>
		<li>Monitör renk değişikliği sülfürik asit düzenli aralıklarla ekleme süre şişeye tepki karışımı. Bir ara ile 4-5 mL damla H2/ çok korumak4; karışım beyaz bir bulutlu çözüm olmalıdır. Not: Anlık sarı yağış konsantre sülfürik asit hızlı ek nedeniyle görünebilir.</li>
		<li>Reaksiyon çözüm adım 1.1.4 başlangıcından bu yana 1 h için ayakta olun.</li>
	</ol>
</li>
	<li>
DMMTAV çıkarma sıvı-sıvı ekstraksiyon yöntemiyle
	<ol>
<li>1 saat sonra yaklaşık 200 mL Dietil eter içeren bir HCI'yi huni tepki solüsyonu dökün.</li>
		<li>5-10 dk, gaz stopcock geçerek birkaç kez serbest bırakmak için huni sallamak. Not: Sentezlenmiş DMMTAV Dietil eter (0.713 g·mL-1) üst katmana aktarılır. Uyarı: Dietil eter gaz zararlı olabilir; iyi havalandırılmış duman başlığı kullan.</li>
		<li>Bir ölçek tepki çözüm toplamak ve ayrı ayrı DMMTAV bir şişe içeren Dietil eter toplamak. Reaksiyon çözüm aynı HCI'yi hunide geri yerleştirin ve yaklaşık 200 mL taze Dietil eter reshaking için ekleyin. Adımları 1.2.2 - 1.2.3 üç kere tekrar edin.</li>
		<li>Tekrar toplanan Dietil eter adım 1.2.3 aynı HCI'yi huni içine dökün ve yaklaşık 100 mL N2ekleyin-deiyonize suyla temizlendi. Sallamak için 5-10 dk ve N2atın-deiyonize su ve bir kaç mL Dietil eter saflık amaçlar için temizlendi. Cam petri kabına (en az 160 mm iç çapı) ve minimum yüksekliği 50 mm içinde kalan Dietil eter toplamak.</li>
		<li>Cam petri kabına bir N2 atmosfer tür dönüştürme önlemek için torpido aktarın. Uyarı: Çözücü vakum pompa ile çıkış pass kutusunun içine çekti değil emin olun.</li>
		<li>Dimethylmonothioarsinate beyaz bir çökelti kadar kuru (crystalized DMMTAV) cam petri kabına üzerinde oluşturulur. Not: Protokol burada duraklatılmış.</li>
	</ol>
</li>
	<li>
Doğrulama sentezlenmiş DMMTAV ve depolama
	<ol>
<li>Crystalized DMMTAV, beyaz çökelti al ve ölçmek ve toplam ağırlığı kaydetmek. Uyarı: Hidrojen sülfür gazı zehirlenmesinden önlemek için iyi havalandırılan duman hood veya torpido kullanın.</li>
		<li>Erime crystalized DMMTAV N250 ml-deiyonize suyla temizlendi ve sarı çökelti 0.2 µm şırınga filtre ile filtre.</li>
		<li>DMAV itibariyle toplam DMMTAV, Yani≈9, 649 mg As· dönüştürülür varsayıyorum L-1. DMMTAV ≈1 mg· hisse senedi çözüm sulandırmak L-1 ve ≈40 µg· L-1 Electrospray iyonlaşma kullanarak doğrulama analizi için kitle Spectromtery (ESI-MS) ve HPLC-ICP-MS, anılan sıraya göre.</li>
		<li>M/z DMMTAV m/z 155 pozitif iyon modunda veya m/z 153 (Tablo 1) negatif iyon modunda ESI-MS11,16 ve parçaları kullanarak analiz. Not: Bkz: başvuru değerleri m/z (Tablo 1).</li>
		<li>DMMTAV HPLC-ICP-MS11,16,17 uygun eluent koşullar ile kullanarak hisse senedi çözümde Kromatografik analiz etmek ve büyük bir tepe'de açıklanan tutma zaman bulundu onaylayın Edebiyat. Not: Sentezlenmiş DMMTAV saflığı adım 1.3.5 analiz sonuçlarından kullanılarak hesaplanması.</li>
		<li>ICP-MS asit sindirim11 ' den sonra kullanarak toplam arsenik konsantrasyonu analiz ve doğru DMMTAV konsantrasyon sentezlenmiş DMMTAV hisse senedi çözüm seyreltme faktörler ve saflık, olduğu gibi aşağıdaki eşitliği kullanarak hesaplar: Analiz toplam konsantrasyonu (µg· olarak L-1) · Seyreltme faktörü · Saflık (%) doğru DMMTAV konsantrasyon DMMTAV hisse senedi çözüm (µg· = L-1)</li>
		<li>Daha fazla nicel Türleşme analiz18için karanlıkta DMMTAV hisse senedi çözüm 4 ° C'de depolayın.</li>
	</ol>
</li>
</ol>2. DMDTAV sentezi
<ol><li>
Kimyasal hazırlama ve molar oranı karıştırma DMAV , Na2S ve H2SO4 Not: DMAV: Na2S:H2kadar4 1:7.5:7.5 =<ol>
<li>DMAV 40 mL deiyonize su 50 mL santrifüj tüpü içinde 1,38 gr geçiyoruz.</li>
		<li>Na2S reaktif 18.01 g Na2S·9H2O bir 250 mL şişe 50 mL deiyonize suda çözülerek hazırlayın.</li>
		<li>4 mL konsantre sülfirik asit (% 96) 40 mL deiyonize su ekleyerek4 reaktif bir 50 mL santrifüj tüpü bulunan H2hazırlamak. Not: DMAVson molar oranı: Na2S:H2SO4 = 1:7.5:7.51,3,4,8,15.</li>
		<li>DMAV çözüm (Adım 2.1.1) hazırlanan 40 mL 50 mL 250 mL şişe (Adım 2.1.2) bulunan Na2S çözeltisi ekleyin. DMAV 10 mL deiyonize su ile içeren tüp durulayın ve bu 250 mL şişe de ekleyin.</li>
		<li>H2hazır 40 mL kadar ekleyin4 çözüm (2.1.3. adım) yavaş yavaş ve kademeli bir şekilde, balonun içine.</li>
		<li>Sülfürik asit düzenli aralıklarla ekleme süre şişeye tepki karışımı renk değişikliği izlemek. Bir ara ile 4-5 mL damla H2/ çok korumak4; karışımı bir beyaz/sarı bulutlu çözüm olmalıdır. Not: Anlık sarı yağış nedeniyle hızlı bir ek konsantre sülfirik asit görünebilir. Uyarı: Sülfürik asit ekleyerek üzerine beyaz duman-ecek var olmak oluşturmak; iyi havalandırılan duman başlığı kullan.</li>
		<li>Flask örtü olmadan gecede tepki çözümde korumak. Not: Protokol burada duraklatılmış.</li>
	</ol>
</li>
	<li>
Katı faz ayıklama (SPE) yöntemini kullanarak DMDTAV çıkarma
	<ol>
<li>Sonra gece ayakta tepki, C18 kullanarak tepki solüsyonu yazın silis tabanlı SPE tuzağa düşürmek için filtre DMDTAV reçine üzerinde sentezledim. Uyarı: İyi havalandırılmış duman başlığı kullan.</li>
		<li>10 mM amonyum asetat (pH 6.3) 0,77 g amonyum asetat deiyonize su 1 litre içinde çözülerek hazırlayın. 10 mM amonyum asetat yeterli hacmi adsorbed DMDTAVayıklamak için C18 şırınga (Adım 2.2.1) elute. Bir cam Petri kabına (en az 160 mm iç çapı) ve minimum yüksekliği 50 mm filtre uygulanmış amonyum asetat toplamak.</li>
		<li>Cam petri kabına bir N2 atmosfer tür dönüştürme önlemek için torpido aktarın. Uyarı: Solvent vakum pompa ile çıkış pass kutusunun içine çekti değil emin olun.</li>
		<li>Dimethyldithioarsinate beyaz bir çökelti kadar kuru (crystalized DMDTAV) cam petri çanak üzerinde kurdu. Not: Protokol burada duraklatılmış.</li>
	</ol>
</li>
	<li>
Doğrulama sentezlenmiş DMDTAV ve depolama
	<ol>
<li>Crystalized DMDTAVbeyaz çökelti al ve ölçüm kayıt toplam ağırlığını ölçmek. Uyarı: Bir iyi havalandırılmış duman hood veya torpido hidrojen sülfür gazı zehirlenmesinden engellemek için kullanın.</li>
		<li>Erime crystalized DMDTAV N250 ml-50 mL tasfiye deiyonize suyla santrifüj kapasitesi tüp ve herhangi bir çökelti 0.2 µm şırınga filtre ile filtre.</li>
		<li>DMAV itibarıyla Toplam DMDTAV, Yani≈2, 539 mg As· dönüştürülür varsayıyorum L-1. DMDTAV ≈1 mg· hisse senedi çözüm sulandırmak L-1 ve ≈40 µg· L-1 ESI-MS ve HPLC-ICP-MS, sırasıyla kullanarak doğrulama analizi için.</li>
		<li>M/z DMDTAV m/z 171 pozitif iyon modunda veya m/z 169 (Tablo 1) negatif iyon modunda ESI-MS11,16 ve parçaları kullanarak analiz. Not: Bkz: başvuru değerleri m/z (Tablo 1).</li>
		<li>DMDTAV HPLC-ICP-MS11,16,17 uygun eluent koşullar ile kullanarak hisse senedi çözümde Kromatografik analiz etmek ve açıklanan tutma zamanı bir tepe bulunabilir doğrulayın Edebiyat. Not: Sentezlenmiş DMDTAV saflığı adım 2.3.5 analiz sonuçlarından kullanılarak hesaplanması.</li>
		<li>ICP-MS asit sindirim11 ' den sonra kullanarak toplam arsenik konsantrasyonu analiz ve doğru DMDTAV konsantrasyon sentezlenmiş DMDTAV hisse senedi çözüm seyreltme faktörler ve saflık aşağıdaki eşitliği kullanarak hesaplar: Analiz toplam konsantrasyonu (µg· olarak L-1) · Seyreltme faktörü · Saflık (%) DMDTA hisse senedi çözüm (µg·V konsantrasyon gerçek DMDTAV = L-1)</li>
		<li>Daha fazla nicel Türleşme analiz18için karanlıkta DMDTAV hisse senedi çözüm 4 ° C'de depolayın.</li>
	</ol>
</li>
</ol>

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Toxicol. 26, 952-962 (2013).</li><li>Alava, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HPLC-ICP-MS+method+development+to+monitor+arsenic+speciation+changes+by+human+gut+microbiota.">HPLC-ICP-MS method development to monitor arsenic speciation changes by human gut microbiota.</a> Biomed. Chromatogr. 26, 524-533 (2012).</li><li>Kurosawa, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+metabolic+activation+associated+with+glutathione+in+dimethylmonoarsinic+acid+(DMMTAV)-induced+toxicity+obtained+from+in+vitro+reaction+of+DMMTAV+with+glutathione.">A novel metabolic activation associated with glutathione in dimethylmonoarsinic acid (DMMTAV)-induced toxicity obtained from in vitro reaction of DMMTAV with glutathione.</a> J. trace Elem. Med. Biol. 33, 87-94 (2016).</li></ol>

Yazarlar ifşa gerek yok.

Gelişmiş iletişim kuralı önceki çalışmalar1,3,4,8,15 atlanmış veya kısaltılmış, hangi zorluklarla veya başarısızlık sırasında yol açmış olabilir önemli adımlar açıklık vardır DMMTAV ve DMDTAV sentezi. DMMTAV oksidasyon duyarlı1,5olduğu iç...

Dimethylarsinic asit (DMAV) Akut toksisite ve metilasyonu ve thiolation yenmesi1,2üzerine geçiren nedeniyle genotoxicity sergilemek için kanıtlanmıştır beri arsenik thiolation metabolik yolu vardır yoğun olmuştur vitro ve in vivo3,4 de olduğu gibi çevre medya (Örneğin, çöp sızıntı)5,6okudu. Önceki yıllarda yapılan çalışmalarda her ikisi de azaltılmış ve thiolated analogları DMAV yaşayan hücreleri, örnek, dimethylarsinous asit (DMAIII), dimethylmonothioarsinic asit (DMMTAV) ve dimethyldithioarsinic asit (için bulduk DMDTAV)7,8,9, dimethylated thioarsenicals DMMTAV diğer inorganik veya organik arsenicals10bilinen daha büyük toksisite sergilenmesi gibi. Son derece sulfidic koşulları11altında çevre ve insanlar için bir risk oluşturabilecek bu yana yüksek derecede toksik thioarsenicals bolluk ciddi çevresel etkileri vardır. Ancak, DMMTAV ve DMDTAV (trans) oluşumu ve onların kaderi çevre medya mekanizmaları hala daha fazla çalışma gerektirir. Böylece, thioarsenicals kantitatif analiz DMMTAV ve DMDTAVçevresel etkileri anlayış geliştirmek için gereklidir.
Standart kimyasallar kantitatif analiz için anahtar gereksinimi olmakla birlikte, DMMTAV ve DMDTAV standartları sayesinde diğer türler türler dönüşmesi riski yüksek önceki çalışmalarda kopyalayan tarafından elde etmek zordur ve unstandardized sentez yordamlar12. Ayrıca, başvurulan yöntemleri standart kimyasallar sentezleme ve kantitatif analiz performans, pratik sorunlara neden olabilir bazı kısıtlamalara sahiptir. DMMTAV ve DMDTAV sık DMAV, Na2S ve H24 ' te bir belirli molar oranı1 veya köpüren H2S gaz Yani DMAV 13,14bir çözüm ile karıştırılarak hazırlanır. Oksijen kullanarak doğrudan temini, son derece zehirli ve zor bir deneyimsiz kullanıcı için denetime H2S gazı kükürt tarafından kabarcıklanma yöntemi özellikleri değiştirme. Tersine, thiolation, DMMTAV ve DMDTAV çevre sudies5,6,12, kalitatif analiz için yaygın olarak kullanılan yukarıdaki karıştırma yöntem1, özellikleri DMAV H2Na2S ve H24 ve üretir DMMTAV ve DMDTAVolarak hedef kimyasallar, üretmek daha kolay stokiometrik kontrol sağlayan, doğrudan karşılaştırıldığında bu yüzden karıştırarak oluşturulan S H2S gaz kullanın.
Yöntemi yordamları1,3,4,8,15 karıştırma başvuru bu çalışma sergi kısıtlamalar bazı yol açabilir kritik onların deneysel adımlar bahsedilen deneysel hatası. Örneğin, belirli solvent (deiyonizeYani, su) hazırlık ve ayıklama ayrıntılarını ve sentezlenmiş arsenicals kristalleşme aşırı kısaltılmış veya yeterince ayrıntılı bir biçimde anlatılan değil. Böyle dağınık ve yordam adımları hakkında sınırlı bilgi thioarsenicals ve güvenilmez miktar analiz tutarsız oluşumuna neden olabilir. Bu nedenle, burada geliştirilen değiştirilmiş protokolünü DMMTAV ve DMDTAV hisse senedi çözümleri sayısal türler ayırma analizi ile sentezi açıklar.

<table width="712">
	<tbody>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Cacodylic acid</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">20835-10G-F</td>
			<td style="width:179px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Sodium sulfide nonahydrate</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">S2006-500G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Sulfuric acid 96%</td>
			<td style="width:179px;">J.T.Baker</td>
			<td style="width:179px;">0000011478</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Ammonium acetate</td>
			<td style="width:179px;">Sigma-Aldrich</td>
			<td style="width:179px;">A7262-500G</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Formic acid 98%</td>
			<td style="width:179px;">Wako Pure Chemical Industries, Ltd.</td>
			<td style="width:179px;">066-00461</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Diethyl ether (Extra Pure)</td>
			<td style="width:179px;">Junsei Chemical</td>
			<td style="width:179px;">33475-0380</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Adapter cap for 60 mL Bond Elut catridges</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">12131004</td>
			<td>Syringe type of SPE</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">Bond Elut C18 cartridge</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">14256031</td>
			<td>Syringe type of SPE</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:179px;height:21px;">HyPURITY C-18</td>
			<td style="width:179px;">Thermo Scientific</td>
			<td style="width:179px;">22105-254630</td>
			<td>5 um, 125 x 4.6 mm</td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Glovebox</td>
			<td style="width:179px;">Chungae-chun, Rep. of Korea</td>
			<td style="width:179px;"></td>
			<td>Customized&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Agilent 1260 Infinity Bio-inert LC</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">DEAB600252, DEACH00245</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="width:179px;height:42px;">Agilent Technologies 7700 Series ICP-MS</td>
			<td style="width:179px;">Agilent Technologies</td>
			<td style="width:179px;">JP12031510</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="width:179px;height:63px;">Finnigan LCQ Deca XP MAX Mass Spectrometer System</td>
			<td style="width:179px;">Thermo Electron Corporation</td>
			<td style="width:179px;">LDM10627</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of dmmtav and dmdtav using dmav for environmental applications: synthesis, purification, and confirmation

Dimethylmonothioarsinic asit (DMMTAV) gibi Dimethylated thioarsenicals ve dimethyldithioarsinic asit (DMDTAV), dimethylarsinic asit (DMAV) thiolation metabolik yolu tarafından üretilen, son zamanlarda olmuştur çevre yanı sıra insan organlarını buldum. DMMTAV ve DMDTAV dimethylated thioarsenicals ve çevre medya onların istikrar ekolojik etkileri belirlemek için sayısal. Bu bileşiklerin sentezi yöntemi önceki çalışmalarda zorlu çoğaltılıyor yapma unstandardized özeldir. Ayrıca, tür dönüşümü olmadan bileşiklerin depolama da dahil olmak üzere depolama teknikleri hakkında bilgi eksikliği vardır. Ayrıca, sadece sınırlı bilgi sentez yöntemleri hakkında kullanılabildiğinden, standart kimyasallar sentezleme ve kantitatif analiz gerçekleştirme deneysel zorluklar olabilir. Burada sunulan protokolü için dimethylated thioarsenicals, DMMTAV ve DMDTAV, pratik olarak değiştirilmiş sentez yöntemi sağlar ve tür ayırma analizi yüksek performanslı sıvı miktar içinde yardımcı olacaktır Kromatografi İndüktif eşleşmiş plazma kütle spektrometresi (HPLC-ICP-MS) ile birlikte. Bu yordamın deneysel adımını kimyasal reaktifler, filtrasyon yöntemleri ve depolama hazırlanması üzerinde odaklanarak tarafından modifiye edilmiştir.

DMMTAV yanlışlıkla DMAIII sentez yöntemi19tarafından hazırlanmış olup, doğrulama sentezlenmiş DMMTAV ve DMDTAV sentezi ve çıkarma ve ideal standart belirleme için önemli bir adım olduğu için kimyasal maddeler. Sentezlenmiş kimyasallar DMMTA en yüksekV (MW 154 g·mol-1) ve DMDTAV (MW 170 g·mol-1) kitle ücret oranı (m/z) electrospray iyonlaşma kütl...

Watch this Scientific Journal Video about Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation at JoVE.com

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

DMMTAV ve çevresel uygulamaları için DMAV kullanarak DMDTAV hazırlanması: sentez, arıtma ve onay

Bu yazı için dimethylmonothioarsinic asit (DMMTAV) değiştirilmiş deneysel protokoller sunar ve dimethyldithioarsinic asit (DMDTAV) sentezi, dimethylarsinic asit (DMAV) thiolation DMAV karıştırma yoluyla inducing , Na2S ve H2kadar4. Değiştirilen protokol böylece deneysel hataları kantitatif analiz sebep olabilecek sentez adımları sınırlamaları aşmak deneysel bir kılavuz sağlar.

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

preparation-dmmtav-dmdtav-using-dmav-for-environmental-applications

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

5.5K Görüntüleme.  Korea Basic Science Institute, Seoul Center. Bu yazı için dimethylmonothioarsinic asit (DMMTAV) değiştirilmiş deneysel protokoller sunar ve dimethyldithioarsinic asit (DMDTAV) sentezi, dimethylarsinic asit (DMAV) thiolation DMAV karıştırma yoluyla inducing , Na2S ve H2kadar4. Değiştirilen protokol böylece deneysel hataları kantitatif analiz sebep olabilecek sentez adımları sınırlamaları aşmak deneysel bir kılavuz sağlar....

Video: Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Waste Water Derived Electroactive Microbial Biofilms: Growth, Maintenance, and Basic Characterization

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup controlled by a potentiostat. Thereby the use of potentiostats allows an exact adjustment of the electrode potential and ensures reproducible microbial culturing conditions. During growth the current production is monitored using chronoamperometry (CA). Based on these data the maximum current density (jmax) and the coulombic efficiency (CE) are discussed as measures for characterization of the bioelectrocatalytic performance. Cyclic voltammetry (CV), a nondestructive, i.e. noninvasive, method, is used to study the extracellular electron transfer (EET) of electroactive bacteria. CV measurements are performed on anodic biofilm electrodes in the presence of the microbial substrate, i.e. turnover conditions, and in the absence of the substrate, i.e. nonturnover conditions, using different scan rates. Subsequently, data analysis is exemplified and fundamental thermodynamic parameters of the microbial EET are derived and explained: peak potential (Ep), peak current density (jp), formal potential (Ef) and peak separation (&#916;Ep). Additionally the limits of the method and the state-of the art data analysis are addressed. Thereby this video-article shall provide a guide for the basic experimental steps and the fundamental data analysis.

Chronoamperometric growth of anodic electroactive microbial biofilms in a fed-batch reactor using a three-electrode setup, controlled by a potentiostat, is demonstrated. The extracellular electron transfer is characterized using cyclic voltammetry in the presence of the electron donor and in the absence of the electron donor. Fundamental data analysis is demonstrated.

Büyüme, Bakım ve Temel Karakterizasyonu: Atık Su Elektroaktif mikrobiyel filmlerin Türetilmiş

The growth of anodic electroactive microbial biofilms from waste water inocula in a fed-batch reactor is demonstrated using a three-electrode setup ...

Çevre

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.

A whole cell bioreporter assay with Burkholderia sartisoli RP037-mChe was developed to detect fractions of an organic contaminant (i.e., fluorene) available for bacterial degradation after active transport by mycelia bridging air-filled pores in a water unsaturated model system.

A Whole Hücre Bioreporter Yaklaşımı Su Doymamış Sistemleri Organik Kontaminantların Taşıma ve Biyoyararlanımını Değerlendirmek için

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

Hazırlık ve Test zehirlilik için hızlı değerlendirme İçme Suyu Örnekleri için RTgill-W1 Hücreleri ile akışkan bioçipler Empedans tabanlı

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in nutrient and gas cycles. After cores with intact sediment-water interfaces are collected, they are submerged in incubation tanks and kept under aerobic conditions at in situ temperatures. To initiate a time course of overlying water chemistry, cores are sealed without bubbles using a top cap with a suspended stirrer. Time courses of 4-7 sample points are used to determine the rate of sediment water exchange. Artificial illumination simulates day-time conditions for shallow photosynthetic sediments, and in conjunction with dark incubations can provide net exchanges on a daily basis. The net measurement of N2 is made possible by sampling a time course of dissolved gas concentrations, with high precision mass spectrometric analysis of N2:Ar ratios providing a means to measure N2 concentrations. We have successfully applied this approach to lakes, reservoirs, estuaries, wetlands and storm water ponds, and with care, this approach provides valuable information on biogeochemical balances in aquatic ecosystems.

The Benthic Exchange of O2, N2 and Dissolved Nutrients Using Small Core Incubations

Here, we present small core incubations for the measurement of sediment-water gas and solute exchange. These will provide reliable measurements of sediment-water exchange that assess the role of sediment in influencing biological and biogeochemical processes in aquatic ecosystems.

O Bentik Değişim<sub&gt; 2</sub&gt; K<sub&gt; 2</subKüçük Çekirdek KUluçkalar Kullanma&gt; ve Çözünmüş Besinler

The measurement of sediment-water exchange of gases and solutes in aquatic sediments provides data valuable for understanding the role of sediments in ...

Use of a Filter Cartridge for Filtration of Water Samples and Extraction of Environmental DNA

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies employed disk fiber filters to collect eDNA from water samples, although a number of microbial studies in aquatic environments have employed filter cartridges, because the cartridge has the advantage of accommodating large water volumes and of overall ease of use. Here we provide a protocol for filtration of water samples using the filter cartridge and extraction of eDNA from the filter without having to cut open the housing. The main portions of this protocol consists of 1) filtration of water samples (water volumes &#8804;4 L or &gt;4 L); (2) extraction of DNA on the filter using a roller shaker placed in a preheated incubator; and (3) purification of DNA using a commercial kit. With the use of this and previously-used protocols, we perform metabarcoding analysis of eDNA taken from a huge aquarium tank (7,500 m3) with known species composition, and show the number of detected species per library from the two protocols as the representative results. This protocol has been developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

We describe a protocol for filtration of water samples with a filter cartridge and extraction of environmental DNA (eDNA) without having to cut open the housing to remove the filter. This protocol is developed for metabarcoding eDNA from fishes, but is also applicable to eDNA from other organisms.

Filtrasyon Su Örnekleri ve Çevre DNA Ekstraksiyon için bir Filtre Kartuşu Kullanımı

Recent studies demonstrated the use of environmental DNA (eDNA) from fishes to be appropriate as a non-invasive monitoring tool. Most of these studies ...

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical data that may be used in predictive modelling to forecast evolutionary responses of natural ecosystems to future environmental changes. However, excluding a few exceptional cases, long-term studies are scarce because of logistic difficulties associated with accessing temporal samples. Temporal dynamics are frequently studied in the laboratory or in controlled mesocosm experiments with exceptional studies that reconstruct the evolution of natural populations in the wild.
Here, a standard operating procedure (SOP) is provided to revive or resurrect dormant Daphnia magna, a widespread zooplankton keystone species in aquatic ecosystems, to dramatically advance the state-of-the-art longitudinal data collection in natural systems. The field of Resurrection Ecology was defined in 1999 by Kerfoot and co-workers, even though the first attempts at hatching diapausing zooplankton eggs date back to the late 1980s. Since Kerfoot&#39;s seminal paper, the methodology of resurrecting zooplankton species has been increasingly frequently applied, though propagated among laboratories only via direct knowledge transfer. Here, an SOP is described that provides a step-by-step protocol on the practice of resurrecting dormant Daphnia magna eggs.
Two key studies are provided in which the fitness response of resurrected Daphnia magna populations to warming is measured, capitalizing on the ability to study historical and modern populations in the same settings. Finally, the application of next generation sequencing technologies to revived or still dormant stages is discussed. These technologies provide unprecedented power in dissecting the processes and mechanisms of evolution if applied to populations that have experienced changes in selection pressure over time.

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

Diriliş uyuyan Daphnia magna: Protokolü ve uygulamaları

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

Kompozisyon ve Bioaerosols farklı çevresel koşullar altında dağıtım Analizi

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the local to federal levels. The continuing evolution of PFAS chemistries presents a challenge to the environmental monitoring, where ongoing development of targeted methods necessarily lags the discovery of new chemical compounds. There is a need, therefore, to have forward-looking methodologies that can detect emerging and unexpected compounds, monitor these species over time, and resolve details of their chemical structure to enable future work in human health. To this end, non-targeted analysis by high-resolution mass spectrometry offers a broad base detection approach that can be combined with almost any sample preparation scheme and provides significant capabilities for compound identification after detection. Herein, we describe a solid-phase extraction (SPE) based sample concentration method tuned for shorter chain and more hydrophilic PFAS chemistries, such as per fluorinated ether acids and sulfonates, and describe analysis of samples prepared in this fashion in both targeted and non-targeted modes. Targeted methods provide superior quantification when reference standards are available but are intrinsically limited to expected compounds when performing analysis. In contrast, a non-targeted approach can identify the presence of unexpected compounds and provide some information about their chemical structure. Information about chemical features can be used to correlate compounds across sample locations and track abundance and occurrence over time.

Here, we present a protocol for the sequential targeted quantification and non-targeted analysis of fluorinated compounds in water by mass spectrometry. This methodology provides quantitative levels of known fluorochemical compounds and identifies unknown chemicals in related samples with semi-quantitative estimates of their abundance.

Ücret belirlenmesi- ve Polyfluorinated kimyasal türler ile kombine hedef ve hedef tarama olmayan yüksek çözünürlüklü kütle spektrometresi iş akışı

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the ...

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term effects are very important in judging their consequences in the environment and on human health. In demonstrating long-term influences, effects of chemicals over generations in recent studies provide new insight. Here, we describe protocols for studying effects of chemicals over multiple generations using free-living nematode Caenorhabditis elegans. Two aspects are presented: (1) trans-generational (TG) and (2) multi-generational effect studies, the latter of which is separated to multi-generational exposure (MGE) and multi-generational residual (MGR) effect studies. The TG effect study is robust with a simple purpose to determine whether chemical exposure to parents can result in any residual consequences on offspring. After the effects are measured on parents, sodium hypochlorite solutions are used to kill the parents and keep the offspring so as to facilitate effect measurement on the offspring. The TG effect study is used to determine whether the offspring are affected when their parent is exposed to the pollutants. The MGE and MGR effect study is systematical used to determine whether continuous generational exposure can result in adaptive responses in offspring over generations. Careful pick-up and transfer are used to distinguish generations to facilitate effect measurement on each generation. We also combined protocols to measure locomotion behavior, reproduction, lifespan, biochemical and gene expression changes. Some example experiments are also presented to illustrate the trans- and multi-generational effect studies.

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants

Trans- and multi-generational effects of persistent chemicals are essential in judging their long-term consequences in the environment and on the human health. We provide novel detailed methods for studying trans- and multi-generational effects using free-living nematode Caenorhabditis elegans.

Toxicants Trans ve çok jenerasyonel etkileri okumak için Caenorhabditis elegans kullanma

Information about toxicities of chemicals are essential in their application and waste management. For chemicals at low concentrations, the long-term ...

Modeling the Size Spectrum for Macroinvertebrates and Fishes in Stream Ecosystems

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological community or food web. Importantly, the size spectrum assumes that individual size, rather than species&#8217; behavioral or life history characteristics, is the primary determinant of abundance within an ecosystem. Thus, unlike traditional allometric relationships that focus on species-level data (e.g., mean species&#8217; body size vs. population density), size spectra analyses are &#8216;ataxic&#8217; &#8211; individual specimens are identified only by their size, without consideration of taxonomic identity. Size spectra models are efficient representations of traditional, complex food webs and can be used in descriptive as well as predictive contexts (e.g., predicting responses of large consumers to changes in basal resources). Empirical studies from diverse aquatic ecosystems have also reported moderate to high levels of similarity in size spectra slopes, suggesting that common processes may regulate the abundances of small and large organisms in very different settings. This is a protocol to model the community-level size spectrum in wadable streams. The protocol consists of three main steps. First, collect quantitative benthic fish and invertebrate samples that can be used to estimate local densities. Second, standardize the fish and invertebrate data by converting all individuals to ataxic units (i.e., individuals identified by size, irrespective of taxonomic identity), and summing individuals within log2 size bins. Third, use linear regression to model the relationship between ataxic M and D estimates. Detailed instructions are provided herein to complete each of these steps, including custom software to facilitate D estimation and size spectra modeling.

This is a protocol to model the size spectrum (scaling relationship between individual mass and population density) for combined fish and invertebrate data from wadable streams and rivers. Methods include: field techniques to collect quantitative fish and invertebrate samples; lab methods to standardize the field data; and statistical data analysis.

Akış ekosistemlerinde Makroomurgasızlar ve balıklar için boyut spektrumunu modelleme

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological ...

DMMTA^V ve çevresel uygulamaları için DMA^V kullanarak DMDTA^V hazırlanması: sentez, arıtma ve onay

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